Examples of the aims to conduct impurity studies [20, 23, 25, 26, 27].
\r\n\tThere will be a chapter on secondary causes of sexual dysfunction disorders related to diabetes, cardiovascular disease, and obesity. A chapter on remedial measures to enhance sexual activity and maintain human relationships will be discussed. As there is a growing number of cancer survivors a chapter on cancer-related sexual dysfunction will be welcomed for including it.
",isbn:null,printIsbn:null,pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,isNomenclature:!1,hash:"b988fda30a4e2364ee9d47e417bd0ba9",bookSignature:"Dr. Dhastagir Sultan Sheriff",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11889.jpg",keywords:"Sex, Sexual Response Cycle, Erection, Premature Ejaculation, Libido, Orgasm, Painful Intercourse, Psychological, Female, Lack of Desire, Erectile Disorders, Pain Disorders",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"April 8th 2022",dateEndSecondStepPublish:"May 6th 2022",dateEndThirdStepPublish:"July 5th 2022",dateEndFourthStepPublish:"September 23rd 2022",dateEndFifthStepPublish:"November 22nd 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"3 months",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dhastagir Sultan Sheriff is a life member of the European Society for Human Reproduction and Early Human Development, Association of Physiologists and Pharmacologists of India, member of the National Academy of Medical Sciences, New Delhi, and resource person for UNESCO for Medical and Bioethics. 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\nImpurities in drug substance (i.e., active pharmaceutical ingredient, API) or drug product can arise due to synthetic/manufacturing processes, degradation, storage conditions, container, excipients, or contamination. They can be identified or unidentified, volatile or nonvolatile, organic or inorganic species [1, 2, 3].
\nSince different regulatory requirements and management strategies are required to be established and complied, sources of impurities shall be carefully classified prior to take subsequent steps; for instance, to distinguish an impurity which is simply derived from API alone or actually derived from interaction products of API-excipient, excipient-excipient, or API-residual impurities existing in excipients [4, 5, 6].
\nDespite an increase in the research of impurities, a number of problems are still arisen in the development of identification technologies for degradation products and pathways. The first aim of this research is to address a brief review of the current major international regulatory requirements regarding the management of impurities in pharmaceutical products. Then secondly, a general scheme to establish an analytical method and acceptance criteria of degradation-related impurities (DRIs) and process-related impurities (PRIs) can be proposed, accordingly. Finally, our research will focus on developing a practicable algorithm to perform the identification of DPIs by using high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS). Meanwhile, verification method for the justification of reliabilities regarding identification results will be assessed.
\nAccording to the definitions of International Council for Harmonization (ICH), Food and Drug Administration (FDA), and USP, impurities are classified into DRIs, PRIs, residual solvents, and heavy metals as shown in Figure 1 [1, 2, 7].
\nClassification of impurities [
Two types of impurities might be API-related. The first type of API-related impurities is generated by degradation of API itself under specific storage conditions, e.g., oxidation, dehydration, carbon dioxide removal, etc. The other type is occurred due to the interaction between API and excipients, container, or residual impurities in excipients, reagents, or solvents [8, 9]. API-related impurities are potentially genotoxic, mutagenic, and carcinogenic risk due to their structure-activity relationship (SRA) [10, 11].
\nIt is well known that excipients or the residual impurities in excipients can be very likely to cause instability of the API and drug product. A lot of impurities in excipients, such as presence of reactive peroxides or high water content in povidone or polyethylene glycols (PEGs), antioxidants in magnesium stearate, aldehydes in lactose, benzaldehyde in benzyl alcohol, formaldehyde in starch, lignin and hemicelluloses in microcrystalline cellulose were illustrated to demonstrate how reactive chemical entities are commonplace in excipients and incompatible to API. Some specific functional groups in API may be susceptible to degradation mechanisms, i.e., hydrolysis, oxidation, polymerization, etc. [4, 5, 6, 12, 13, 14].
\nAdditionally, extractables and leachables such as initiators/catalysts, storage stabilizers, antioxidants, processing aids, light stabilizers, antistatic agents, colorants, lubricants associated with pharmaceutically relevant materials may also produce uncertain risks to the stability or quality of products [15].
\nRegardless of the classes of impurities, presence of impurities may have the potential to affect the quality, safety, and efficacy of drug products. Therefore, studies of impurities are one of the most important works in the development of APIs and drug products [1, 16, 17].
\nStudy of impurities in pharmaceuticals is one of the most highly regarded topics; it is essential, but time consuming and challenging. In terms of regulations and technology, we must keep pace with the times [18, 19]. Comprehensively speaking, aims to develop an impurity study have two major directions as follows: regulatory requirements and scientific/technical demands (Table 1).
\nRegulatory requirements | \nScientific and technical requirements | \n
---|---|
\n
| \n\n
| \n
From the perspective of regulatory requirements, impurities may affect the quality of APIs and DPs and ultimately affect the safety of the patient. Views for the dealing of impurities may differ between biologists, toxicologists, and analytical chemists, and therefore need to be integrated [20]. Potential genotoxic impurities can be determined according to the published literature, results of gene mutation in bacteria, in vitro and in vivo tests of chromosomal damage in mammalian cells or rodent hematopoietic cells, or/and comparative structural analysis to identify chemical functional moieties correlated with mutagenicity [16]. Moreover, daily exposure, duration of exposure on the effects of degradation products and genotoxic impurities, and theoretical clinical dose, whereas potential mutagenic impurities must be controlled to levels less than the threshold of toxicological concern based on lifetime exposure shall be evaluated as a risk consideration [16, 17, 18].
\nAdequate qualification must include genotoxicity and repeat-dose toxicology studies of appropriate duration to support the proposed indication. Moreover, other specific toxicity studies, e.g., embryofetal developmental toxicity study may be appropriate. Genotoxic impurities and degradation products pose an additional risk and should be controlled in accordance with the requirements of ICH M7(R1), unless they are qualified for safety [18, 21].
\nIn addition to the regulatory requirements, internal and external scientific and technical needs are the second perspective to conduct an impurity study. Impurity determination and forced degradation studies are two of the basic requirements as a tool to predict potential DPIs, to develop analytical method, synthetic processes, and formulation, to receive a better understanding of storage conditions, stability of drug product, and to obtain information of degradation products/pathways, as well as to evaluate the specificity (selectivity) of assay method [22, 23, 24, 25].
\nA number of international/local guidelines and guidances for the evaluation and control of impurities in drug substances and drug products have been published [1, 2, 3, 7, 8, 9, 21, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38]. Comparison of the application scopes in line with the impurity categories was drawn as indicated in Figure 2.
\nComparison of the application scopes of regulatory guidelines/guidance for the management of impurities in pharmaceutical products [
As said by the requirements of ICH Q3A(R2), all types of impurities present in API at a level greater than (>) the identification threshold must conduct studies to characterize their structures, no matter they are shown in any batch manufactured by the proposed commercial process or any degradation product observed in stability studies under recommended storage conditions. Specified identified impurities shall be included in the list of impurities along with specified unidentified impurities that are estimated to be present at a level greater than the identification threshold [2, 7, 33].
\nBriefly, five major steps for the management of degradation products, no matter they are degradation products of API or reaction products of API with excipient(s) or container closure system, have been requested by the ICH Q3B (R2) and summarized as follows [3]:
Confirm which impurities are degradation products?
Monitor and/or specify the amount of all degradation products.
Summarize all degradation products during manufacture and stability studies.
Elucidate and justify a rational evaluation of possible degradation pathway in the drug product or interaction with excipients or container closure system.
Establish specifications of all degradation products, including specified identified, specified unidentified, unspecified degradation product with an acceptance criterion of not more than (≤) identification threshold described in Q3B (R2), and their total amount.
Specificity (selectivity) of the method applied to determine specified and unspecified degradation product shall be validated. This includes subjecting of API or drug products to stress studies of light, heat, humidity, acid and base hydrolysis, and oxidation to evaluate the HPLC separation resolution, mass balance, etc. [3, 22, 24, 25].
\nAlthough Q3B (R2) was developed by ICH to provide guidance on impurities in drug products for new drug applications (NDAs), it is also considered to be applicable to the drug products of abbreviated new drug application (ANDAs) [33].
\nRegulation requirements regarding genotoxic, mutagenic, and carcinogenic impurities have been published and revised by European Medicine Agency (EMA), FDA, and ICH in 2006, 2008, and 2017, respectively, to describe how to perform assessments and controls, including prevention and reduction of impurities [21, 28, 32].
\nConcept of threshold of toxicological concern (TTC) has been developed to define an acceptable intake for any unstudied chemical that poses a negligible risk of carcinogenicity or other toxic effects [21]. In general, exposure level of 1.5 μg per person per day (i.e., TTC) for each impurity can be considered as a common acceptable qualification threshold for supporting marketing application. Any impurity found at a level below this threshold generally does not need further safety qualification for genotoxicity and carcinogenicity concerns. The threshold is an estimate of daily exposure expected to result in an upper-bound lifetime risk of cancer of less than 10−6 (one in a million), a risk level that is thought to pose negligible safety concerns [21, 32].
\nCurrently, ICH Q3C is the major guideline related to the management of residual solvents in API, excipients, and drug products (Figure 2). In general, solvents that are used in the manufacturing procedures are the required parts to determine [8]. Types of solvents are sorted according to their carcinogenic and genotoxic risks as follows [8, 37]:
Class 1: solvents obviously confirmed or strongly suspected to cause cancer in humans.
Class 2: nongenotoxic and possible carcinogenic risks in animals.
Class 3: low-toxic solvents.
Elemental impurities may arise from residual catalysts that were added intentionally in synthesis, or may be present as impurities, e.g., through interactions with processing equipment or container/closure systems or by being present in components of the drug product. Because elemental impurities pose toxicological concerns and do not provide any therapeutic benefit to the patient, their levels in drug products should be controlled within acceptable limits. Appropriate documentation demonstrating compliance for detailed risk assessment, screenings, and validation data for release methods must be conducted [9, 30, 34].
\nRecommended maximum acceptable concentration limits for the residues of metal catalysts or metal reagents that may be present in pharmaceutical products were issued earlier by EMA [29, 30]. Another classification of impurities, i.e., elemental impurities that the pharmaceutical industry needs to comply with is defined recently in ICH Q3D [9]. Comparison for these classifications of residues of metal or elemental impurities in pharmaceutical products defined by EMA and ICH was indicated as shown in Figure 3. Several significant difference of elemental safety concerns between EMA and ICH, such as Cr, As, Cd, Hg, Pb, etc., can be found.
\nComparison for the classification of residues of metal and elemental impurities in pharmaceutical products by requirements of EMA and ICH Q3D [
This chapter will be followed by a discussion of procedure to establish an analytical method and acceptance criteria of DRIs and PRIs.
\nSteps for the determination of potential degradation products, including a science-based risk assessment, can been addressed as below [11, 25]:
Stress studies of API.
Accelerated stability studies or kinetically equivalent shorter term stability studies.
Validation/verification by long-term stability studies of both the drug substance and formulated drug product.
An integrated scheme in accordance with the requirements of ICH for the establishment of analytical methods and acceptance criteria of PRIs and DRIs is proposed as demonstrated in Figure 4 [2, 3, 17, 22, 39, 40].
\nScheme to establish analytical methods and acceptance criteria of process-related impurities (PRIs) and degradation-related impurities (DRIs) according to the requirements of ICH guidelines [
In general, when an unknown peak was found, no matter it was found in a stress or stability studies of API or drug product, the first step is to distinguish the classification of unknown impurity belongs to. Different regulatory requirements of the management for different kinds of impurities, i.e., PRIs and DRIs are required to apply. For instance, requirements of ICH Q3B(R2) and Q1A(R2) request that impurities present in API need not be monitored or specified in the drug product unless they are also degradation products. Due to the probability of degradation during storage period and are likely to influence quality, safety, and/or efficacy, degradation impurities must be included into the plan of stability studies [39]. Meanwhile, degradation impurities can ultimately determine the expiration, retest, or shelf-life periods of API and drug products, by evaluating the intersection of extrapolation-upper confidence limit and upper acceptance criterion of degradation product(s) [40]. Reporting threshold, identification threshold, and qualification threshold in the case of maximum daily dose ≤2 g/day of APIs administrated are illustrated in Figure 4 [17].
\nStructure of impurities present in API at a level greater than (>) the identification threshold needs to be elucidated. An identified impurity content can be either determined by interpolation with calibration curve of reference material or calculated using the peak area of the main component, i.e., API. In contrast, unidentified impurity content can only be determined using the peak area of API, no matter they are specified or unspecified impurities. Impurities with specific acceptance criteria are referred to as specific impurities, including identified and unidentified impurities [2].
\nBefore conducting method validation, all of the impurities shall be verified by spiked or known addition to demonstrate they do exist under the “real” storage conditions such as accelerated or long-term storage conditions. Otherwise, it may not be necessary to examine specifically for certain degradation products if they are not formed under the “real” storage conditions [11, 25, 39].
\nThe method for technology transfer to QC laboratory, i.e., receiving unit (RU) must be a well-validated and stability-indicating method. A method fails to pass the criteria of validation or technology transfer, investigation to clarify the root cause(s) and revalidation shall be initiated and conducted by the originating unit (OU) and approved by quality unit (QU).
\nAlgorithms for the identification and verification of DRIs are proposed as indicated in Figure 5. Degradation reaction kinetics can be represented by a linear regression curve on an arithmetic or logarithmic scale [39]. Meanwhile, nature of degradation relationship is determined by the reaction kinetic constants and can be accordingly used to distinguish whether an impurity is DRI or PRI compound (Figure 5).
\nAlgorithms for the identification and verification of API-related degradation impurities (DRIs).
One example regarding how to distinguish PRIs and DRIs by kinetic study was illustrated as demonstrated in Figure 6. Analysis by HPLC revealed that some impurities were existed in one of our products. Kinetic study helps us to distinguish the type of impurities.
\nKinetic study of impurities formation by conducting stress studies to distinguish DRIs and PRIs.
Plots of the impurity formation concentration ([A] or Ln[A]) versus time can obtain rate constant, i.e., the slope of a reaction in straight line as arithmetic (i.e., k0) or logarithmic (i.e., k1) scale. Furthermore, correlation coefficient (r) of linear regression analysis indicates a perfect positive correlation (r = 1) or conversely, there is no relationship between the two variables (r = 0).
\nThe slopes and correlation coefficients of Pk#5, Pk#6, and Pk#7 indicated that they were not degradation-related products of API. But conversely, kinetic curves showed that Pk#1–4 and Pk#8 were degradation products. These results were also consistent with the findings of molecular weight results shown in LC-MS/MS (data not shown).
\nAs shown in Figure 5, the first step for structure elucidation is running full Q1 scans in both positive ion mode and negative ion mode to locate the m/z of parent peak. In this step, sample solution is typically introduced directly into mass spectrometer (MS) at a flow rate of 10 μL/min using a syringe pump. However, since dimer or oligomer may also be one of the potential impurities, range of Q1 scan shall be as wide as possible, e.g., to mass number of 1000–1200 at least.
\nCarefully compare the difference of mass-to-charge (∆m/z) numbers between experimental and nominal values of parent (molecular) peak as well as their stable isotope distribution patterns and natural abundances. Previous study for the elucidation of degradation pathways of ethyl cysteinate dimer (ECD), a significant ∆m/z value of −2 in Q1 scan between experimental result (m/z = 323.60) and nominal result (m/z = 325.46) of parent peak was found and indicating that an intramolecular disulfide (S-S) product, i.e., [ECDS-S+H]+, was the prominent form of ECD (not [ECD+H]+) in aqueous solution before labeling of radioisotope, i.e., technetium-99m for i.v. injection (Figure 7) [25].
\nStructures of (a) ethyl cysteinate dimer (ECD), (b) intramolecular disulfide (S-S) product of ECD, i.e., ECDS-S, and (c) intermolecular dimer of ECD and reducing agent (SnCl2), i.e., Sn(ECD)2 (DP#4) [
Repeat the product ion scans, precursor ion scans, and neutral loss scans of API to establish its collision-activated dissociation (CID) fragmentation database, including the optimal CID energies of each fragment and multiple reaction monitoring (MRM) pairs. Propose the promising structures of CID fragments and fragmentation pathways of API, accordingly. Provide the comparison of ∆m/z results between experimental and nominal values for each peak, which is related to the fragmentation to verify the reliability of proposed fragments and fragmentation pathways [24, 25].
\nLinear relationship within dynamic ranges for the quantitation of MRM pairs, i.e., correlation coefficients (r = 1) between precursor ions and product ions is another indication to verify high stability and reproducibility of fragmentation in CID conditions of tandem MS [24, 25].
\nBefore using the MRM pairs for impurity scanning, interference of fragments generated from background, matrix, or contaminants such as plasticizers present in the solvents and mobile phase must be verified. Plasticizers, e.g., di(2-ethylhexyl) phthalate (DEHP) are one of the most common contaminants in organic solvents, including acetonitrile and alcohol [41].
\nRepeat the same procedures mentioned above in Figure 5 to obtain a comprehensive information of fragments for any available intermediates and degradation products which are received from synthetic division, from contract manufacturing organization (CMO), from a stress study, or stability study sample conducted by the R&D team.
\nSteps for the determination of impurities related to degradation of API are illustrated as follows:
Step 1: According to the CID fragments of API, intermediates, or/and degradation products, a list of potential core fragments, which may be related to the unknown component(s) is proposed.
Step 2: Predict a set of potential/extending MRM pairs in line with the list obtained in step 1 and then coupled it with the relevant (bio-) transformations under the storage conditions of APIs/drug products for conducting MS/MS scans.
Step 3: Conduct the precursor ion scans together with function of information-dependent acquisition (IDA), where CID is automatically performed on the two highest intensity MS peaks to find the possible precursor ions containing core fragments established in step 2.
Step 4: Perform the reliability assessment by analysis commercial batches or long-term/accelerated stability samples to verify the identification results of step 3.
One preliminary study was illustrated as shown in Figure 8 can be used to detail the algorithms of Figure 5. Core fragment of m/z 243 was found in the MS/MS study of API. In the meantime, four potential extending core fragments, i.e., m/z 183, m/z 185, m/z 197, and m/z 199 were obtained by the MS/MS studies of intermediate and degradation product (Step 1).
\nStep for the establishment of potential extending core fragments, conduct of product ions screening with transformation/IDA function, and validation/verification.
A total of five potential core fragments, coupled with the experience accumulated by degradation products that may be produced by similar chemical structures and prediction of relevant (bio-) transformations reactions under storage conditions, such as oxidation (+O, +2O), dehydration (−H2O, −2(H2O)), remove of carbon dioxide, and remove of acetic acid, a set of MRM pairs for scanning is established (Step 2).
\nConduct the precursor ion scans by coupled with the IDA function for automatic performing collision on the two highest intensity MS peaks in the targeting regions of HPLC (Step 3). (Note: IDA is a build-in function of API 4000 QTrap (AB Sciex) for conducting an automatic collision on the highest intensity peak(s) scan.)
\nIn addition to the methods mentioned above, i.e., kinetic study and difference of mass-to-charge (∆m/z) between experimental and nominal results, three other evaluation methods to verify the reliability of the identification results are available: including verification by real samples, by stable isotope distribution patterns, and by mass balance.
Verification by real samples
Investigation results of unknown degradation product(s) must be verified by the “real samples”, i.e., commercial batches or long-term/accelerated stability studies samples. Verification of reliability is achieved by comparison the difference of retention time (tR), MRM pairs, and stable isotope distribution patterns between real samples and stress study samples. If it is available, purified or enrichment sample of impurity can be spiked into a real sample for further verification.
2. Verification by stable isotope distribution patterns or natural abundances
Each element, like a fingerprint, has its own unique stable isotope distribution patterns and natural abundances. Occasionally, stable isotope distribution patterns or natural abundances are available as a unique tool for structure characterization.
\nTen, two, and two of uncommon patterns in the MS spectra as shown in Figure 9(a)–(c) were clearly indicated in our structure identification of ethyl cysteinate dimer (ECD) cold kit, (R)-N-methyl-3-(2-bromophenoxy)-3-phenylpropanamine (MBPP), and methoxyisobutylisonitrile (sestamibi, or Cu(MIBI)4), respectively. These uncommon patterns were attributed to the contribution of stable isotope distributions of tin (Sn), bromine (Br), and copper (Cu), respectively.
\nStable isotope distribution patterns and simulation of mass spectra of (a) Sn(ECD)2 (DP#4), (b) (R)-N-methyl-3-(2-bromophenoxy)-3-phenylpropanamine (MBPP), and (c) methoxyisobutylisonitrile (sestamibi).
When 7 major (or actually total 10) peaks are shown in the MS spectra, it may strongly mislead the works of structure elucidation as shown in Figure 9(a). However, if it is available to know the presence of some special elements may present in impurity.
\nIf it is able to presuppose that some special elements may contain in the structure, then it will be easier to elucidate the MS spectra. In other words, when pattern of MS spectra is significantly different from the normal CHO distribution, it may also indicate that a special element exists on the structure.
\nBy comparing the natural abundance of 10 stable isotopes of tin and simulation MS spectra of a promising molecular formula, a series of metal complexes of tin can be verified. In the case for study of impurities in ECD kit, it was an ultimate and effective way to identify all of impurities containing Sn, i.e., DP#4, DP#5, DP#6, DP#6′, DP#6″, DP#7, DP#7′, DP#7″, and DP#8 [25]. Similar case was found in the structure determination of sestamibi as shown in Figure 9(c). Coordination number (CN = 4) and core metal (Cu) in sestamibi can be clearly verified.
3. Verification by mass balance
When performing a stress study of API, one should determine content of API on each day by using a daily and freshly prepared calibration curve of API reference material, and interpolated within the validated dynamic range. The mass balance is calculated by summation of the API and total impurity content. It is a tool to justify whether there are impurities unseparated (i.e., same retention time) or undetectable (e.g., without UV-visible chromophores). This topic and several major problems to cause poor mass balance have been detailed by Nussbaum et al. [42]
\nManagement of impurities related to APIs in pharmaceutical products must be implemented in strict compliance with the regulatory requirements of pharmaceutical industry due to their quality and safety concerns. An integrated scheme in accordance with the regulatory requirements to establish analytical methods and acceptance criteria of process-related impurities (PRIs) and degradation-related impurities (DRIs) was presented, accordingly. Meanwhile, procedures for the identification and validation/verification of API-related DRIs were proposed. Validation or verification methods to evaluate the reliability of structure identification such as kinetic reactions, stress and stability studies, comparison of retention time(s) and ∆m/z between experimental and nominal values of targeting peaks, compatibility of MRM pairs with “real samples,” stable isotope distribution patterns, and mass balance were demonstrated. Applying of the processes proposed in this article will help to ensure the reliability and quality of the impurity analytical results.
\nThe authors would like to thank Dr. Shyh-Fong Chen, Mr. Chu-Huang Hsieh, Miss Jane-Yu Huang, Miss Pi-Lin Liu, and Mr. Yung-Hsuan Sung, Pharmaceutical Business Unit, Everlight Chemical Industrial Corporation (ECIC) for their helpful advice on the Regulatory Requirements in this chapter. The authors also appreciate Dr. Lai-Chuan Chang, Biotech Total Solutions Co., Ltd., Dr. Lee-Chung Men, Dr. Lie-Hang Shen, Dr. Mei-Hsiu Liao, Miss Yi-Chih Hsia, and Mr. Chang-Yung Su, Institute of Nuclear Energy Research (INER) for their great supporting on the radiopharmaceutical and analytical works.
\nDairy farming plays an essential role in the global socioeconomic scenario, being one of the most traditional rural activities and fundamental for agribusiness. Milk is one of the most complete and most consumed food globally, in addition to providing a social function, generating thousands of direct and indirect jobs throughout the production chain. The development of new technologies should add more efficiency to the milk production chain, a constant challenge for several sectors involved in the segment.
The use of animal reproduction biotechnologies has contributed to the increase in animal productivity and has been one of the main responsible for the increase in milk production. The current scenario is the search for a model capable of providing high production efficiency concerning animal welfare without harming the environment and with the most advanced reproductive techniques for obtaining pregnancies and genetic improvement. In this way, ovarian physiology is a key aspect to contribute to the efficiency of dairy production.
The ovarian follicular population is characterized by the total amount of follicles present in the ovary. Each follicle contains an oocyte, so it is known that there are a great number of oocytes in the ovary. However, only a small portion of the ovarian follicles undergo ovulation. Therefore, the ovarian follicular reserve is an important indicator of fertility in cattle, which may influence the applicability of reproductive biotechniques.
This chapter addresses the mechanism of folliculogenesis and the most recent research. It also brings discussions on how reproductive biotechniques can influence fertility in dairy cattle.
The origin of the female reproductive system is still in embryonic life in the sublumbar region located caudally to the kidneys. The primordial germ cells, which will give rise to the germline formation, originate in the proximal epiblast and then move from the yolk sac to the gonadal ridges through the mesentery, around day 30 of germinal development [1]. At this moment, the Müller and Wolff ducts are still present, which will give rise to the female and male reproductive tract, respectively.
After the colonization of the ridges, around 35 days of gestation, the differentiation process begins by specifying the somatic cells of the ridge, where Sertoli cells will originate from the XY chromosome, and the granulosa cells will originate from the XX chromosome. There is an involution of Wolff’s duct (or mesonephros) and development of Müller’s duct (or paramesonephric) in the escarpments. In males, as they inherit the testicles determining factor (TDF) from the Y chromosome, Sertoli cells release the anti-müllerian hormone (AMH) and inhibit the development of Müller’s ducts [2, 3]. After this process, they are formed as oogonia that through mitotic and meiotic divisions form a nest of oogonia in a tubular shape, and then a process of differentiation into oocytes begins [4].
Gonadal structures called germline cysts are elevated in the ovigerous cords and surrounded by a basement membrane shortly after colonization of the gonadal ridges by primordial germ cells. Meiotic divisions are initiated until the process is stopped in meiosis prophase I when primary oocytes are already formed. The primary oocytes are surrounded by a layer of undifferentiated pregranulosa cells [5]. The interruption of meiosis can last for years until a given follicle enters the growth process, resuming meiosis and continuing the follicular development through the primordial follicle until its final stage in the antral follicle [6].
It is known that folliculogenesis depends on interactions between the somatic cells of the follicle and the oocyte, so the communication between the granulosa and theca cells with the oocyte is essential for follicular development and growth to occur [7, 8]. The passage from the primordial follicle to the primary follicle is a transition phase and is characterized by the action of specific growth factors for each stage of folliculogenesis [9]. In bovine species, the so-called follicular growth waves correspond to a stimulus for the recruitment of preantral follicles.
Once the primordial follicle is recruited, whose granulosa cells are flat, it becomes the primary follicle and there is a transition between the flat cells to the cuboidal-shaped granulosa cells [9]. At this stage, the zona pellucida appears, which will remain around the oocyte throughout the follicle’s development. Continuing to grow, the secondary follicle is constituted when the granulosa cells multiply and form two layers of cubic morphology, in addition to the emergence of the first theca cells [10].
The growth of these secondary follicles (when they reach approximately 4 mm in diameter) is regulated by the follicle-stimulating hormone (FSH), which has its receptors in the granulosa cells. When they reach a larger size (approximately 7–9 mm in diameter), they start to be controlled by luteinizing hormone (LH). At this stage, the follicle is already characterized as tertiary and has LH receptors in the theca cells that are already entirely organized, and the formation of the follicular antrum can be observed [11, 12]. The phases of follicular development are shown in Figure 1.
Schematic sequence of complete follicular development.
The follicular population may vary between individuals, and some factors such as genetics, breed, age, species, and hormone levels can influence the number of follicles present in the ovaries [13]. For bovine females, it is estimated that the number of follicles at birth is about 235,000 [14].
An increase in the number of antral follicles present in the ovary and stimulated by gonadotrophin secretion is influenced by body development in heifers [15]. Endocrine activity at first seems to be controlled by suppressing negative feedback mechanisms until the heifer has a good body condition to initiate the estrous cycle and reproduction activities [16, 17, 18].
The first ovulation in heifers is marked by a gradual increase in LH secretion, which leads to the development of the antral follicles and the secretion of estrogen. At birth, antral follicles are not typically present in the ovaries, and the number of follicles increases when heifers reach 2 months of age. After that, the number of follicles declines at 5 months of age, and some changes continue to occur throughout the productive life of the female [19, 20, 21].
Ovarian follicular population is highly variable among species, a concept that is well established [20, 21, 22] and has been already reported in cattle [23]. In recent years, numerous studies have focused on the ovarian follicular population and its influence on reproductive activities, as well as on animal reproduction biotechniques [22, 24]. The antral follicle count (AFC) is a strategy to identify different profiles of cows, performing transrectal B-mode ultrasonography and counting all follicles larger than 3 mm [25]. The total number of follicles counted in the pair of ovaries is added up, and the cow is classified as low, medium, or high AFC. A feature in cattle is the high variability of AFC between animals, but it is known that there is high repeatability in the same individual [19, 26, 27]. The appearance of the ovary on ultrasound examination of cows with high and low AFC is presented in Figure 2.
Aspect of the ovary on ultrasound examination of cows with different counts of antral follicles (AFC). On the right (A) cow with low AFC, and the left (B) cow with high AFC. Images were generated via the transrectal route with the equipment model S8v (SonoScape®) with a frequency of 8.6 MHz and a linear transducer of 5–10.0 MHz.
Furthermore, according to [28], it has already been established that the concentrations of anti-müllerian hormone (AMH), which is released by the granulosa cells of growing ovarian follicles, are positively related to the entry into puberty of bovine females. In other words, the higher serum concentrations of this hormone are, the higher AFC will be [27]. AMH is a glycoprotein that belongs to the TGF-β growth factor family [29] and it is correlated with follicular growth [30]. AMH is in the granulosa cells and it is responsible for the growth of preantral and antral follicles [31] and follicular growth modulator through the control of ovarian follicular reserve depletion [32].
The intrafollicular AMH expression increases until the follicle reaches 5 mm in cows and then decreases as the follicle reaches the antral stage and increases in size [31]. The positive correlation of AMH with the ovarian follicular population has already been described in previous studies [33, 34]. Thus, the measurement of AMH can be a method of predicting AFC [35]. In
Reproductive biotechniques, such as embryo transfer (ET) and IVEP, depends on the population of antral follicles present in the ovary of donor females to succeed. Among other factors that interfere with ET and IVEP, it is important to mention genetics, breed, and age [37, 38]. High AFC bovine females have been described to have a greater number of viable embryos produced
In contrast, a high conception rate was observed after the use of TAI in low AFC females
Donors with a high number of antral follicles have been selected, mainly for OPU-
In summary, several studies have tried to verify the influence and the correlation of AFC in the reproductive performance of bovine females. So far, it is not possible to establish the role of AFC in fertility parameters due to the controversial results. Although it is quite predictable that AFC may be related to reproductive efficiency, a better understanding of the subject is necessary. Furthermore, considering basic research, it is necessary to elucidate some aspects of follicular physiology that remain unknown [47].
Increasing the productive efficiency of a herd is one of the great challenges for dairy cattle farming. In the past, genetic selection programs sought essential characteristics for increasing milk production, with effective gains in milk quantity and quality, but reproductive efficiency was disregarded. In recent years, several works have been presented to increase milk production and increase reproductive performance, a key association for efficient dairy farming.
Considering the importance of a sustainable, intensive, and economically viable production system, achieving the reproductive efficiency of the dairy herd is crucial for the effects on profitability by the number of offspring produced, genetic progress, and the shorter interval between lactations. This is a great challenge, as there is low heritability between production and reproduction traits. Therefore, the crucial importance of precisive reproductive assistance is highlighted, providing maximum production efficiency in the smallest possible area and respecting the aspects of animal comfort.
Despite the rapid development of the technique since its emergence in the late 1980s, until recently, IVEP was used only as a last resource when traditional techniques failed. However, the high genetic gains provided to the herds, obtaining a greater number of pregnancies concerning
Holstein cows typically have lower oocyte production when subjected to IVEP. However, it is possible to obtain good results by performing a pre-selection of females with a high number of antral follicles using ultrasound. It is important to highlight that non-lactating females often have a higher number of follicles and oocytes.
Until a few years ago, some obstacles prevented the large-scale use of IVEP in dairy cattle. One of them was the large number of calves born from unwanted sex (male), which significantly increased the production cost. Another difficulty was the distance, often thousands of kilometers, between the laboratories and the properties where the recipient cows were located. The inefficiency of cryopreservation techniques for IVP embryos, especially when dealing with
These two major obstacles have now been overcome, making large-scale
In addition, some alternatives can be employed to improve the methods that assist in the recovery of better-quality oocytes and a higher competence in OPU to obtain more interesting results in embryo production. In this context, the follicular wave synchronization before OPU and consider the influence of the antral follicle population seem to be good alternatives [50].
Dairy European breeds, such as Holstein and Jersey, suffer great discomfort under high temperatures and high humidity conditions. Therefore, failures in cyclicity and the demonstration of estrus occur, making management difficult and compromising AI results. Furthermore, it is known that embryos are naturally more resistant to heat stress than gametes, which can suffer degeneration and further reduce pregnancy rates in the summer [51]. The transfer of embryos 7 days after fertilization avoids the harmful effects before this period, providing more advantageous rates than AI [52, 53, 54]. For dairy cattle, therefore, the use of transferred embryos seems to be the most viable option, especially in periods of excessive heat [55].
In AI, the number of descendants of genetically selected bulls is multiplied. IVEP, in turn, also generates descendants of females of high genetic merit, causing an even more significant impact on the improvement of a herd. Greater genetic gain is achieved in each generation with the transfer of embryos produced
In some dairy farms, cows with better genetic potential are used as embryo donors and recipients. Thus, an efficient genetic selection from animals in the herd becomes possible. As for the economic aspect, with the number of pregnancies in IVEP, it is possible to produce embryos at affordable costs, making the embryo commercially attractive compared to semen [56]. Another advantage is the better use of high-value semen due to the possibility of fertilizing ten or more cows with a single dose.
The use of conventional semen, both in AI and
The most used technique for semen sexing is flow cytometry, which offers an accuracy of 85–95% [59]. However, during the sexing process, the sperm may be damaged, which might compromise their viability, reducing the fertilization potential and embryonic development [60]. The sexing process reduces sperm motility, compromising AI indices. [61] Related that the mean conception rate after AI between 2012 and 2016 was 56.9% with conventional semen and 47.3% using sexed semen. In IVEP, however, the method allows obtaining very satisfactory rates of blastocysts, with quality similar to those produced with conventional semen, since this technique requires fewer viable spermatozoa [62, 63]. Generally, the conception rate obtained with sexed semen is 50–60% of the rates obtained with conventional semen in cows and 70–90% of conventional semen in heifers [64].
As there is a reduction in fertility using this semen, some strategies are currently suggested to improve conception rates in insemination programs that use sexed semen. First, it is recommended to use this semen in heifers and most in the first three services due to greater fertility. In AI programs with heat observation, the highest conception rates were achieved, with AI being performed between 16 and 24 h after the onset of heat [65]. Finally, in TAI programs, the best rates were achieved with semen deposition 60 h after removing the progesterone source [66].
In the
The
The production of embryos by SOV also does not allow pregnant cows, while in IVEP, this is possible. Follicular aspiration can be performed as long as the ovaries can be manipulated without being subjected to excessive traction. The process flow of
Schematic sequence of steps in the
Schematic sequence of steps in the
The cryopreservation of bovine embryos generated
The greater sensitivity of these embryos to low temperatures is mainly due to the greater accumulation of lipids in the cytoplasm [71]. Lipids, made up mostly of triacylglycerols, directly affect the survival of embryos during cooling, as they can undergo irreversible changes and severely compromise development. An alternative method to promote chemical delipidation of embryos and increase cryotolerance by decreasing lipid accumulation has been related [72]. Forskolin, for example, a compost derived from the Indian plant
Among the cryopreservation methods, vitrification is the most used technique worldwide due to the speed of the process and its low cost [74]. On the other hand, direct transfer (DT), a technique used to simplify the
The
Due to the metabolic and morphological differences compared to those produced
Thus, if the proposal is to associate embryo transfer and sexed semen, the best strategy at the moment is the
In order to minimize the effects of early embryonic loss, the Doppler ultrasound technique has been included in reproductive programs. This non-invasive and real-time biotechnology allows the characterization of blood perfusion of reproductive organs and tissues throughout the estrous cycle and pregnancy in cattle. One of its purposes is to accurately estimate the corpus luteum (CL) functionality for the selection of recipients and for the early diagnosis of pregnancy in TAI and TETF (Fixed Time Embryo Transfer) programs.
In addition to allowing for greater accuracy in the evaluation of the recipient, another feature of the Doppler is the diagnosis of pregnancy at 20–22 days, which is early compared to the conventional system performed at 30 days after insemination. Super-early resynchronization programs developed in heifers and cows are being introduced in dairy herds, as the reduction in the interval between two TAIs promotes gains in reproductive efficiency. Despite the correct evaluation being dependent on the experience and knowledge of the operator and the correct configuration of the equipment, the popularization of the technique is consolidated every day and presents good prospects for the future.
The current scenario of reproduction biotechnology demonstrates great potential for a sustainable increase in milk production, mainly due to the increase in reproductive and productive efficiency. Furthermore, the growth in the use of reproductive biotechniques is associated with the parallel development of a support network such as veterinarians, the pharmaceutical industry, disposable materials, equipment, and service providers. The generation of employment and the need to train human resources to meet the demand for activities are intended to provide social growth.
With the possibility of obtaining an accelerated genetic gain through the shortening of the generation interval, the use of prepubertal females, mainly in the production of embryos, has aroused great commercial interest and investment in research. The genetic potential of the female must first be evaluated in advance, that is, before total production. This is feasible thanks to progress in research with genetic markers for accurate prediction of the females that will be more efficient in milk production. It is also important to consider improving equipment for OPU (oocyte recovery by Ovum pick-up). There are currently fully adapted transducers for use in very young females. Despite the good number of aspirated follicles, a challenge in this category is the low blastocyst rate, promoting limited results in IVF.
Thus, to be viable for the use of these females, the next step is to develop protocols that improve the competence of the retrieved oocyte. Gonadotropin stimulus to increase the proportion (and size) of large follicles and synchronization of follicular waves before OPU to decrease immature oocytes have been investigated. A revolution in dairy farming that has become increasingly accessible is genomic selection which has significantly altered the global dairy industry. The reduction in the generation interval from 7 to 2.5 years and the reduction of costs with progeny tests were only the first benefits presented by the gene-editing biotechnique.
Silencing, altering or replacing genes that cause problems are effective strategies to increase the productive efficiency of the herd, selecting and breeding genetically superior animals. The generation gap is likely to narrow further as assessments gain wide acceptance, as genetic gains are cumulative across generations. Genetic progress is expected as continued genetic selection is implemented. Since 2009, more than one million animals have received genetic evaluations. Although these tests are carried out primarily on male animals, genotyping costs are currently economically viable. Currently, genomic selection programs are investing more in health traits (resistance to disease), reproduction, and selection for environmentally sustainable production, including reducing waste production and gas emissions.
This change of concept, which seeks longevity and animal welfare, is because, in recent years, there has been a decline in fertility and resistance in several populations, leading to a decrease in the profitability of the herds. The increase in slaughter rates, veterinary expenses, replacement costs, and reduced milk sales were just some of the consequences of the negative impact of years of selection focusing only on milk production and animal appearance. Furthermore, the adoption of a selection index, such as evaluating the quality and viability of embryos before the transfer, increases the efficiency of the process.
An example of this has been in North America, where the implementation of a genetic-based selection program for reproductive disorders is actively researched. A high and positive genetic correlation between retained placenta and metritis is being observed, implying selection of genes to improve one trait reflecting positively on the other. This demonstrates that the increased need for genomic traits for these traits contributes to the reproductive efficiency of dairy herds.
Other characteristics that have been valued in genomic tests are identifying biomarkers considered for genetic improvement, highly correlated with reproductive performance, such as anti-müllerian hormone (AMH), and identifying relevant genes to reduce pregnancy losses. Identifying genetic markers related to the development and anticipation of the embryo and their selection to avoid embryonic losses can minimize economic damage. Another issue to be further elucidated shortly is whether genes relevant to embryonic development are positively associated with fertility traits. Estimates of the heritability of conventional reproductive traits are generally low. Even so, the progressive inclusion of genomic tests, as a routine in the field, has great potential for identifying superior animals. In the medium and long term, one perspective is that genetic improvement programs will bring consistent profitability for the dairy industry.
Genomic testing still faces challenges because a decisive outcome in the short term is unlikely. Genetic variation for economic characteristics is maintained by increased frequency of rare alleles, new mutations and changes in goals, and no selection management. Moreover, although genomic selection is being well applied at rates of genetic gain, we still know very little about the genetic structure that promotes this variation. The most relevant future challenge will probably be the incorporation of new characteristics in the selection index in breeding programs, overcoming a measurement difficulty or low heritability of them. Added to this, it is still uncertain whether traits produced over several generations emerged included in routine genomics, as gene frequencies change over time.
It is already known that the selection of some genes can directly or indirectly influence other aspects. The concern with creations called “ecologically correct” remains controversial. The inclusion of characteristics such as lower gas emissions can compromise herd productivity. It should be remembered that the increase in milk production per animal reduces the total production of residues in the atmosphere. In other words, it is something broader than simply a genetic alteration to favor an environmental issue narrowly.
Genomic testing positively changes productivity dynamics, but attention is needed to the consequences of these genetic manipulations. The pioneering application of genomic selection in cattle will lead to a series of unanticipated discoveries that could affect animals and society. An accidental finding was recently published in highly relevant research. It was discovered that two cloned bulls whose cell lineage had undergone gene editing, aiming at the characteristic of not having horns, were transgenic. The animals contained in their genome the genetic material of the bacterium used as a vector in gene editing. The Food and Drugs Administration (FDA) guarantees that intentional genomic alterations are safe for animals and anyone who consumes foods derived from them. However, there is still no universally accepted verification method for genomic editing.
Finally, with all the technological changes, the dairy herd has its premises, but the consumer market has also increased its requirements. Producers face the challenge that today there are claims for harmonic milk in ingestion (A2A2) [78], welfare for female producers, and respect for the environmental preservation area. People worldwide are looking for information about the products daily and are no longer limited to the final part of the milk production chain.
The increase in reproductive efficiency is a proposal fully adjusted to environmental sustainability. More productive herds require less area to generate more feed. Furthermore, the use of genetically improved animals according to the climatic conditions of each region prevents land competition with agriculture. As for differentiated milk production, the inclusion of bulls genotyped for the A2 allele of beta-casein accelerates aggregation of A2A2 animals in the herd.
Another critical aspect is the mandatory link between reproductive biotechnology and animal welfare. More productive animals only respond to greater reproductive efficiency if they have all vital requirements well met. Technological innovations such as robotic milking, with the cow’s autonomy about milking, signals a prospect of increased milk production with the same number of animals. A new change in concept which, adding welfare to the creation of dairy cattle will reflect positively on the profitability of producers.
All aspects of folliculogenesis remain a vast area to be studied, despite the notable progress made with previous research. It is not possible to determine the complete influence of AFC on female bovine fertility. The use of AFC as a tool to produce embryos
Overcoming the main limitations of IVEP, together with the good results and its high applicability, has contributed to the use of biotechnique on a large scale. Thus, IVEP is no longer limited to elite animals or animals that do not respond to superovulation but actively contributes to the production, improvement, and profitability of dairy production.
The transfer of IVP embryos is a great strategy to reduce the cost of high genetic value semen, and it seems to be the most viable option in periods or regions of high temperatures. Thus, IVEP has benefited dairy farms of all sizes and animals of different breeds, whether
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\n\nCorresponding authors will receive a 25% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters. A 20% discount for publishing a long-form monographs, 25% for compacts and 23% for short-form monographs.
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He is currently a principal researcher in data analytics and optimisation at TECNALIA (Spain), a visiting fellow at the Basque Center for Applied Mathematics (BCAM) and a part-time lecturer at the University of the Basque Country (UPV/EHU). His research interests gravitate on the use of descriptive, prescriptive and predictive algorithms for data mining and optimization in a diverse range of application fields such as Energy, Transport, Telecommunications, Health and Industry, among others. In these fields he has published more than 240 articles, co-supervised 8 Ph.D. theses, edited 6 books, coauthored 7 patents and participated/led more than 40 research projects. 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He is currently a full professor in\nthe Department of Automation and Applied Informatics at the\nsame university. Dr. Voloşencu is the author of ten books, seven\nbook chapters, and more than 160 papers published in journals\nand conference proceedings. He has also edited twelve books and\nhas twenty-seven patents to his name. He is a manager of research grants, editor in\nchief and member of international journal editorial boards, a former plenary speaker, a member of scientific committees, and chair at international conferences. His\nresearch is in the fields of control systems, control of electric drives, fuzzy control\nsystems, neural network applications, fault detection and diagnosis, sensor network\napplications, monitoring of distributed parameter systems, and power ultrasound\napplications. 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Radiotherapy and Nuclear Medicine Technology has always been my aspiration and my life. As years passed I accumulated a tremendous amount of skills and knowledge in Radiotherapy and Nuclear Medicine, Conventional Radiology, Radiation Protection, Bioinformatics Technology, PACS, Image processing, clinically and lecturing that will enable me to provide a valuable service to the community as a Researcher and Consultant in this field. My method of translating this into day to day in clinical practice is non-exhaustible and my habit of exchanging knowledge and expertise with others in those fields is the code and secret of success.",institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"313277",title:"Dr.",name:"Bartłomiej",middleName:null,surname:"Płaczek",slug:"bartlomiej-placzek",fullName:"Bartłomiej Płaczek",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/313277/images/system/313277.jpg",biography:"Bartłomiej Płaczek, MSc (2002), Ph.D. (2005), Habilitation (2016), is a professor at the University of Silesia, Institute of Computer Science, Poland, and an expert from the National Centre for Research and Development. His research interests include sensor networks, smart sensors, intelligent systems, and image processing with applications in healthcare and medicine. He is the author or co-author of more than seventy papers in peer-reviewed journals and conferences as well as the co-author of several books. He serves as a reviewer for many scientific journals, international conferences, and research foundations. Since 2010, Dr. Placzek has been a reviewer of grants and projects (including EU projects) in the field of information technologies.",institutionString:"University of Silesia",institution:{name:"University of Silesia",country:{name:"Poland"}}},{id:"35000",title:"Prof.",name:"Ulrich H.P",middleName:"H.P.",surname:"Fischer",slug:"ulrich-h.p-fischer",fullName:"Ulrich H.P Fischer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/35000/images/3052_n.jpg",biography:"Academic and Professional Background\nUlrich H. P. has Diploma and PhD degrees in Physics from the Free University Berlin, Germany. He has been working on research positions in the Heinrich-Hertz-Institute in Germany. Several international research projects has been performed with European partners from France, Netherlands, Norway and the UK. He is currently Professor of Communications Systems at the Harz University of Applied Sciences, Germany.\n\nPublications and Publishing\nHe has edited one book, a special interest book about ‘Optoelectronic Packaging’ (VDE, Berlin, Germany), and has published over 100 papers and is owner of several international patents for WDM over POF key elements.\n\nKey Research and Consulting Interests\nUlrich’s research activity has always been related to Spectroscopy and Optical Communications Technology. Specific current interests include the validation of complex instruments, and the application of VR technology to the development and testing of measurement systems. He has been reviewer for several publications of the Optical Society of America\\'s including Photonics Technology Letters and Applied Optics.\n\nPersonal Interests\nThese include motor cycling in a very relaxed manner and performing martial arts.",institutionString:null,institution:{name:"Charité",country:{name:"Germany"}}},{id:"341622",title:"Ph.D.",name:"Eduardo",middleName:null,surname:"Rojas Alvarez",slug:"eduardo-rojas-alvarez",fullName:"Eduardo Rojas Alvarez",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/341622/images/15892_n.jpg",biography:null,institutionString:null,institution:{name:"University of Cuenca",country:{name:"Ecuador"}}},{id:"215610",title:"Prof.",name:"Muhammad",middleName:null,surname:"Sarfraz",slug:"muhammad-sarfraz",fullName:"Muhammad Sarfraz",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/215610/images/system/215610.jpeg",biography:"Muhammad Sarfraz is a professor in the Department of Information Science, Kuwait University. 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Prof. Sarfraz is also an editor-in-chief and editor of various international journals.",institutionString:"Kuwait University",institution:{name:"Kuwait University",country:{name:"Kuwait"}}},{id:"32650",title:"Prof.",name:"Lukas",middleName:"Willem",surname:"Snyman",slug:"lukas-snyman",fullName:"Lukas Snyman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/32650/images/4136_n.jpg",biography:"Lukas Willem Snyman received his basic education at primary and high schools in South Africa, Eastern Cape. He enrolled at today's Nelson Metropolitan University and graduated from this university with a BSc in Physics and Mathematics, B.Sc Honors in Physics, MSc in Semiconductor Physics, and a Ph.D. in Semiconductor Physics in 1987. After his studies, he chose an academic career and devoted his energy to the teaching of physics to first, second, and third-year students. After positions as a lecturer at the University of Port Elizabeth, he accepted a position as Associate Professor at the University of Pretoria, South Africa.\r\n\r\nIn 1992, he motivates the concept of 'television and computer-based education” as means to reach large student numbers with only the best of teaching expertise and publishes an article on the concept in the SA Journal of Higher Education of 1993 (and later in 2003). The University of Pretoria subsequently approved a series of test projects on the concept with outreach to Mamelodi and Eerste Rust in 1993. In 1994, the University established a 'Unit for Telematic Education ' as a support section for multiple faculties at the University of Pretoria. In subsequent years, the concept of 'telematic education” subsequently becomes well established in academic circles in South Africa, grew in popularity, and is adopted by many universities and colleges throughout South Africa as a medium of enhancing education and training, as a method to reaching out to far out communities, and as a means to enhance study from the home environment.\r\n\r\nProfessor Snyman in subsequent years pursued research in semiconductor physics, semiconductor devices, microelectronics, and optoelectronics.\r\n\r\nIn 2000 he joined the TUT as a full professor. Here served for a period as head of the Department of Electronic Engineering. Here he makes contributions to solar energy development, microwave and optoelectronic device development, silicon photonics, as well as contributions to new mobile telecommunication systems and network planning in SA.\r\n\r\nCurrently, he teaches electronics and telecommunications at the TUT to audiences ranging from first-year students to Ph.D. level.\r\n\r\nFor his research in the field of 'Silicon Photonics” since 1990, he has published (as author and co-author) about thirty internationally reviewed articles in scientific journals, contributed to more than forty international conferences, about 25 South African provisional patents (as inventor and co-inventor), 8 PCT international patent applications until now. Of these, two USA patents applications, two European Patents, two Korean patents, and ten SA patents have been granted. A further 4 USA patents, 5 European patents, 3 Korean patents, 3 Chinese patents, and 3 Japanese patents are currently under consideration.\r\n\r\nRecently he has also published an extensive scholarly chapter in an internet open access book on 'Integrating Microphotonic Systems and MOEMS into standard Silicon CMOS Integrated circuitry”.\r\n\r\nFurthermore, Professor Snyman recently steered a new initiative at the TUT by introducing a 'Laboratory for Innovative Electronic Systems ' at the Department of Electrical Engineering. The model of this laboratory or center is to primarily combine outputs as achieved by high-level research with lower-level system development and entrepreneurship in a technical university environment. Students are allocated to projects at different levels with PhDs and Master students allocated to the generation of new knowledge and new technologies, while students at the diploma and Baccalaureus level are allocated to electronic systems development with a direct and a near application for application in industry or the commercial and public sectors in South Africa.\r\n\r\nProfessor Snyman received the WIRSAM Award of 1983 and the WIRSAM Award in 1985 in South Africa for best research papers by a young scientist at two international conferences on electron microscopy in South Africa. He subsequently received the SA Microelectronics Award for the best dissertation emanating from studies executed at a South African university in the field of Physics and Microelectronics in South Africa in 1987. In October of 2011, Professor Snyman received the prestigious Institutional Award for 'Innovator of the Year” for 2010 at the Tshwane University of Technology, South Africa. This award was based on the number of patents recognized and granted by local and international institutions as well as for his contributions concerning innovation at the TUT.",institutionString:null,institution:{name:"University of South Africa",country:{name:"South Africa"}}},{id:"317279",title:"Mr.",name:"Ali",middleName:"Usama",surname:"Syed",slug:"ali-syed",fullName:"Ali Syed",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/317279/images/16024_n.png",biography:"A creative, talented, and innovative young professional who is dedicated, well organized, and capable research fellow with two years of experience in graduate-level research, published in engineering journals and book, with related expertise in Bio-robotics, equally passionate about the aesthetics of the mechanical and electronic system, obtained expertise in the use of MS Office, MATLAB, SolidWorks, LabVIEW, Proteus, Fusion 360, having a grasp on python, C++ and assembly language, possess proven ability in acquiring research grants, previous appointments with social and educational societies with experience in administration, current affiliations with IEEE and Web of Science, a confident presenter at conferences and teacher in classrooms, able to explain complex information to audiences of all levels.",institutionString:null,institution:{name:"Air University",country:{name:"Pakistan"}}},{id:"75526",title:"Ph.D.",name:"Zihni Onur",middleName:null,surname:"Uygun",slug:"zihni-onur-uygun",fullName:"Zihni Onur Uygun",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/75526/images/12_n.jpg",biography:"My undergraduate education and my Master of Science educations at Ege University and at Çanakkale Onsekiz Mart University have given me a firm foundation in Biochemistry, Analytical Chemistry, Biosensors, Bioelectronics, Physical Chemistry and Medicine. After obtaining my degree as a MSc in analytical chemistry, I started working as a research assistant in Ege University Medical Faculty in 2014. In parallel, I enrolled to the MSc program at the Department of Medical Biochemistry at Ege University to gain deeper knowledge on medical and biochemical sciences as well as clinical chemistry in 2014. In my PhD I deeply researched on biosensors and bioelectronics and finished in 2020. Now I have eleven SCI-Expanded Index published papers, 6 international book chapters, referee assignments for different SCIE journals, one international patent pending, several international awards, projects and bursaries. In parallel to my research assistant position at Ege University Medical Faculty, Department of Medical Biochemistry, in April 2016, I also founded a Start-Up Company (Denosens Biotechnology LTD) by the support of The Scientific and Technological Research Council of Turkey. Currently, I am also working as a CEO in Denosens Biotechnology. The main purposes of the company, which carries out R&D as a research center, are to develop new generation biosensors and sensors for both point-of-care diagnostics; such as glucose, lactate, cholesterol and cancer biomarker detections. My specific experimental and instrumental skills are Biochemistry, Biosensor, Analytical Chemistry, Electrochemistry, Mobile phone based point-of-care diagnostic device, POCTs and Patient interface designs, HPLC, Tandem Mass Spectrometry, Spectrophotometry, ELISA.",institutionString:null,institution:{name:"Ege University",country:{name:"Turkey"}}},{id:"267434",title:"Dr.",name:"Rohit",middleName:null,surname:"Raja",slug:"rohit-raja",fullName:"Rohit Raja",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/267434/images/system/267434.jpg",biography:"Dr. Rohit Raja received Ph.D. in Computer Science and Engineering from Dr. CVRAMAN University in 2016. His main research interest includes Face recognition and Identification, Digital Image Processing, Signal Processing, and Networking. Presently he is working as Associate Professor in IT Department, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur (CG), India. He has authored several Journal and Conference Papers. He has good Academics & Research experience in various areas of CSE and IT. He has filed and successfully published 27 Patents. He has received many time invitations to be a Guest at IEEE Conferences. He has published 100 research papers in various International/National Journals (including IEEE, Springer, etc.) and Proceedings of the reputed International/ National Conferences (including Springer and IEEE). He has been nominated to the board of editors/reviewers of many peer-reviewed and refereed Journals (including IEEE, Springer).",institutionString:"Guru Ghasidas Vishwavidyalaya",institution:{name:"Guru Ghasidas Vishwavidyalaya",country:{name:"India"}}},{id:"246502",title:"Dr.",name:"Jaya T.",middleName:"T",surname:"Varkey",slug:"jaya-t.-varkey",fullName:"Jaya T. Varkey",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/246502/images/11160_n.jpg",biography:"Jaya T. Varkey, PhD, graduated with a degree in Chemistry from Cochin University of Science and Technology, Kerala, India. She obtained a PhD in Chemistry from the School of Chemical Sciences, Mahatma Gandhi University, Kerala, India, and completed a post-doctoral fellowship at the University of Minnesota, USA. She is a research guide at Mahatma Gandhi University and Associate Professor in Chemistry, St. Teresa’s College, Kochi, Kerala, India.\nDr. Varkey received a National Young Scientist award from the Indian Science Congress (1995), a UGC Research award (2016–2018), an Indian National Science Academy (INSA) Visiting Scientist award (2018–2019), and a Best Innovative Faculty award from the All India Association for Christian Higher Education (AIACHE) (2019). She Hashas received the Sr. Mary Cecil prize for best research paper three times. She was also awarded a start-up to develop a tea bag water filter. \nDr. Varkey has published two international books and twenty-seven international journal publications. She is an editorial board member for five international journals.",institutionString:"St. Teresa’s College",institution:null},{id:"250668",title:"Dr.",name:"Ali",middleName:null,surname:"Nabipour Chakoli",slug:"ali-nabipour-chakoli",fullName:"Ali Nabipour Chakoli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/250668/images/system/250668.jpg",biography:"Academic Qualification:\r\n•\tPhD in Materials Physics and Chemistry, From: Sep. 2006, to: Sep. 2010, School of Materials Science and Engineering, Harbin Institute of Technology, Thesis: Structure and Shape Memory Effect of Functionalized MWCNTs/poly (L-lactide-co-ε-caprolactone) Nanocomposites. Supervisor: Prof. Wei Cai,\r\n•\tM.Sc in Applied Physics, From: 1996, to: 1998, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Determination of Boron in Micro alloy Steels with solid state nuclear track detectors by neutron induced auto radiography, Supervisors: Dr. M. Hosseini Ashrafi and Dr. A. Hosseini.\r\n•\tB.Sc. in Applied Physics, From: 1991, to: 1996, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Design of shielding for Am-Be neutron sources for In Vivo neutron activation analysis, Supervisor: Dr. M. Hosseini Ashrafi.\r\n\r\nResearch Experiences:\r\n1.\tNanomaterials, Carbon Nanotubes, Graphene: Synthesis, Functionalization and Characterization,\r\n2.\tMWCNTs/Polymer Composites: Fabrication and Characterization, \r\n3.\tShape Memory Polymers, Biodegradable Polymers, ORC, Collagen,\r\n4.\tMaterials Analysis and Characterizations: TEM, SEM, XPS, FT-IR, Raman, DSC, DMA, TGA, XRD, GPC, Fluoroscopy, \r\n5.\tInteraction of Radiation with Mater, Nuclear Safety and Security, NDT(RT),\r\n6.\tRadiation Detectors, Calibration (SSDL),\r\n7.\tCompleted IAEA e-learning Courses:\r\nNuclear Security (15 Modules),\r\nNuclear Safety:\r\nTSA 2: Regulatory Protection in Occupational Exposure,\r\nTips & Tricks: Radiation Protection in Radiography,\r\nSafety and Quality in Radiotherapy,\r\nCourse on Sealed Radioactive Sources,\r\nCourse on Fundamentals of Environmental Remediation,\r\nCourse on Planning for Environmental Remediation,\r\nKnowledge Management Orientation Course,\r\nFood Irradiation - Technology, Applications and Good Practices,\r\nEmployment:\r\nFrom 2010 to now: Academic staff, Nuclear Science and Technology Research Institute, Kargar Shomali, Tehran, Iran, P.O. Box: 14395-836.\r\nFrom 1997 to 2006: Expert of Materials Analysis and Characterization. Research Center of Agriculture and Medicine. Rajaeeshahr, Karaj, Iran, P. O. Box: 31585-498.",institutionString:"Atomic Energy Organization of Iran",institution:{name:"Atomic Energy Organization of Iran",country:{name:"Iran"}}},{id:"248279",title:"Dr.",name:"Monika",middleName:"Elzbieta",surname:"Machoy",slug:"monika-machoy",fullName:"Monika Machoy",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/248279/images/system/248279.jpeg",biography:"Monika Elżbieta Machoy, MD, graduated with distinction from the Faculty of Medicine and Dentistry at the Pomeranian Medical University in 2009, defended her PhD thesis with summa cum laude in 2016 and is currently employed as a researcher at the Department of Orthodontics of the Pomeranian Medical University. She expanded her professional knowledge during a one-year scholarship program at the Ernst Moritz Arndt University in Greifswald, Germany and during a three-year internship at the Technical University in Dresden, Germany. She has been a speaker at numerous orthodontic conferences, among others, American Association of Orthodontics, European Orthodontic Symposium and numerous conferences of the Polish Orthodontic Society. She conducts research focusing on the effect of orthodontic treatment on dental and periodontal tissues and the causes of pain in orthodontic patients.",institutionString:"Pomeranian Medical University",institution:{name:"Pomeranian Medical University",country:{name:"Poland"}}},{id:"252743",title:"Prof.",name:"Aswini",middleName:"Kumar",surname:"Kar",slug:"aswini-kar",fullName:"Aswini Kar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/252743/images/10381_n.jpg",biography:"uploaded in cv",institutionString:null,institution:{name:"KIIT University",country:{name:"India"}}},{id:"204256",title:"Dr.",name:"Anil",middleName:"Kumar",surname:"Kumar Sahu",slug:"anil-kumar-sahu",fullName:"Anil Kumar Sahu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/204256/images/14201_n.jpg",biography:"I have nearly 11 years of research and teaching experience. I have done my master degree from University Institute of Pharmacy, Pt. Ravi Shankar Shukla University, Raipur, Chhattisgarh India. I have published 16 review and research articles in international and national journals and published 4 chapters in IntechOpen, the world’s leading publisher of Open access books. I have presented many papers at national and international conferences. I have received research award from Indian Drug Manufacturers Association in year 2015. My research interest extends from novel lymphatic drug delivery systems, oral delivery system for herbal bioactive to formulation optimization.",institutionString:null,institution:{name:"Chhattisgarh Swami Vivekanand Technical University",country:{name:"India"}}},{id:"253468",title:"Dr.",name:"Mariusz",middleName:null,surname:"Marzec",slug:"mariusz-marzec",fullName:"Mariusz Marzec",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/253468/images/system/253468.png",biography:"An assistant professor at Department of Biomedical Computer Systems, at Institute of Computer Science, Silesian University in Katowice. Scientific interests: computer analysis and processing of images, biomedical images, databases and programming languages. He is an author and co-author of scientific publications covering analysis and processing of biomedical images and development of database systems.",institutionString:"University of Silesia",institution:{name:"University of Silesia",country:{name:"Poland"}}},{id:"212432",title:"Prof.",name:"Hadi",middleName:null,surname:"Mohammadi",slug:"hadi-mohammadi",fullName:"Hadi Mohammadi",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/212432/images/system/212432.jpeg",biography:"Dr. Hadi Mohammadi is a biomedical engineer with hands-on experience in the design and development of many engineering structures and medical devices through various projects that he has been involved in over the past twenty years. Dr. Mohammadi received his BSc. and MSc. degrees in Mechanical Engineering from Sharif University of Technology, Tehran, Iran, and his PhD. degree in Biomedical Engineering (biomaterials) from the University of Western Ontario. He was a postdoctoral trainee for almost four years at University of Calgary and Harvard Medical School. He is an industry innovator having created the technology to produce lifelike synthetic platforms that can be used for the simulation of almost all cardiovascular reconstructive surgeries. He’s been heavily involved in the design and development of cardiovascular devices and technology for the past 10 years. He is currently an Assistant Professor with the University of British Colombia, Canada.",institutionString:"University of British Columbia",institution:{name:"University of British Columbia",country:{name:"Canada"}}},{id:"254463",title:"Prof.",name:"Haisheng",middleName:null,surname:"Yang",slug:"haisheng-yang",fullName:"Haisheng Yang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/254463/images/system/254463.jpeg",biography:"Haisheng Yang, Ph.D., Professor and Director of the Department of Biomedical Engineering, College of Life Science and Bioengineering, Beijing University of Technology. He received his Ph.D. degree in Mechanics/Biomechanics from Harbin Institute of Technology (jointly with University of California, Berkeley). Afterwards, he worked as a Postdoctoral Research Associate in the Purdue Musculoskeletal Biology and Mechanics Lab at the Department of Basic Medical Sciences, Purdue University, USA. He also conducted research in the Research Centre of Shriners Hospitals for Children-Canada at McGill University, Canada. Dr. Yang has over 10 years research experience in orthopaedic biomechanics and mechanobiology of bone adaptation and regeneration. He earned an award from Beijing Overseas Talents Aggregation program in 2017 and serves as Beijing Distinguished Professor.",institutionString:null,institution:{name:"Beijing University of Technology",country:{name:"China"}}},{id:"89721",title:"Dr.",name:"Mehmet",middleName:"Cuneyt",surname:"Ozmen",slug:"mehmet-ozmen",fullName:"Mehmet Ozmen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/89721/images/7289_n.jpg",biography:null,institutionString:null,institution:{name:"Gazi University",country:{name:"Turkey"}}},{id:"265335",title:"Mr.",name:"Stefan",middleName:"Radnev",surname:"Stefanov",slug:"stefan-stefanov",fullName:"Stefan Stefanov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/265335/images/7562_n.jpg",biography:null,institutionString:null,institution:{name:"Medical University Plovdiv",country:{name:"Bulgaria"}}},{id:"242893",title:"Ph.D. Student",name:"Joaquim",middleName:null,surname:"De Moura",slug:"joaquim-de-moura",fullName:"Joaquim De Moura",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/242893/images/7133_n.jpg",biography:"Joaquim de Moura received his degree in Computer Engineering in 2014 from the University of A Coruña (Spain). In 2016, he received his M.Sc degree in Computer Engineering from the same university. He is currently pursuing his Ph.D degree in Computer Science in a collaborative project between ophthalmology centers in Galicia and the University of A Coruña. His research interests include computer vision, machine learning algorithms and analysis and medical imaging processing of various kinds.",institutionString:null,institution:{name:"University of A Coruña",country:{name:"Spain"}}},{id:"294334",title:"B.Sc.",name:"Marc",middleName:null,surname:"Bruggeman",slug:"marc-bruggeman",fullName:"Marc Bruggeman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/294334/images/8242_n.jpg",biography:"Chemical engineer graduate, with a passion for material science and specific interest in polymers - their near infinite applications intrigue me. \n\nI plan to continue my scientific career in the field of polymeric biomaterials as I am fascinated by intelligent, bioactive and biomimetic materials for use in both consumer and medical applications.",institutionString:null,institution:null},{id:"255757",title:"Dr.",name:"Igor",middleName:"Victorovich",surname:"Lakhno",slug:"igor-lakhno",fullName:"Igor Lakhno",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/255757/images/system/255757.jpg",biography:"Igor Victorovich Lakhno was born in 1971 in Kharkiv (Ukraine). \nMD – 1994, Kharkiv National Medical Univesity.\nOb&Gyn; – 1997, master courses in Kharkiv Medical Academy of Postgraduate Education.\nPh.D. – 1999, Kharkiv National Medical Univesity.\nDSC – 2019, PL Shupik National Academy of Postgraduate Education \nProfessor – 2021, Department of Obstetrics and Gynecology of VN Karazin Kharkiv National University\nHead of Department – 2021, Department of Perinatology, Obstetrics and gynecology of Kharkiv Medical Academy of Postgraduate Education\nIgor Lakhno has been graduated from international training courses on reproductive medicine and family planning held at Debrecen University (Hungary) in 1997. Since 1998 Lakhno Igor has worked as an associate professor in the department of obstetrics and gynecology of VN Karazin National University and an associate professor of the perinatology, obstetrics, and gynecology department of Kharkiv Medical Academy of Postgraduate Education. Since June 2019 he’s been a professor in the department of obstetrics and gynecology of VN Karazin National University and a professor of the perinatology, obstetrics, and gynecology department. He’s affiliated with Kharkiv Medical Academy of Postgraduate Education as a Head of Department from November 2021. Igor Lakhno has participated in several international projects on fetal non-invasive electrocardiography (with Dr. J. A. Behar (Technion), Prof. D. Hoyer (Jena University), and José Alejandro Díaz Méndez (National Institute of Astrophysics, Optics, and Electronics, Mexico). He’s an author of about 200 printed works and there are 31 of them in Scopus or Web of Science databases. Igor Lakhno is a member of the Editorial Board of Reproductive Health of Woman, Emergency Medicine, and Technology Transfer Innovative Solutions in Medicine (Estonia). He is a medical Editor of “Z turbotoyu pro zhinku”. Igor Lakhno is a reviewer of the Journal of Obstetrics and Gynaecology (Taylor and Francis), British Journal of Obstetrics and Gynecology (Wiley), Informatics in Medicine Unlocked (Elsevier), The Journal of Obstetrics and Gynecology Research (Wiley), Endocrine, Metabolic & Immune Disorders-Drug Targets (Bentham Open), The Open Biomedical Engineering Journal (Bentham Open), etc. He’s defended a dissertation for a DSc degree “Pre-eclampsia: prediction, prevention, and treatment”. Three years ago Igor Lakhno has participated in a training course on innovative technologies in medical education at Lublin Medical University (Poland). Lakhno Igor has participated as a speaker in several international conferences and congresses (International Conference on Biological Oscillations April 10th-14th 2016, Lancaster, UK, The 9th conference of the European Study Group on Cardiovascular Oscillations). His main scientific interests: are obstetrics, women’s health, fetal medicine, and cardiovascular medicine. \nIgor Lakhno is a consultant at Kharkiv municipal perinatal center. He’s graduated from training courses on endoscopy in gynecology. He has 28 years of practical experience in the field.",institutionString:null,institution:null},{id:"244950",title:"Dr.",name:"Salvatore",middleName:null,surname:"Di Lauro",slug:"salvatore-di-lauro",fullName:"Salvatore Di Lauro",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0030O00002bSF1HQAW/ProfilePicture%202021-12-20%2014%3A54%3A14.482",biography:"Name:\n\tSALVATORE DI LAURO\nAddress:\n\tHospital Clínico Universitario Valladolid\nAvda Ramón y Cajal 3\n47005, Valladolid\nSpain\nPhone number: \nFax\nE-mail:\n\t+34 983420000 ext 292\n+34 983420084\nsadilauro@live.it\nDate and place of Birth:\nID Number\nMedical Licence \nLanguages\t09-05-1985. Villaricca (Italy)\n\nY1281863H\n474707061\nItalian (native language)\nSpanish (read, written, spoken)\nEnglish (read, written, spoken)\nPortuguese (read, spoken)\nFrench (read)\n\t\t\nCurrent position (title and company)\tDate (Year)\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. Private practise.\t2017-today\n\n2019-today\n\t\n\t\nEducation (High school, university and postgraduate training > 3 months)\tDate (Year)\nDegree in Medicine and Surgery. University of Neaples 'Federico II”\nResident in Opthalmology. Hospital Clinico Universitario Valladolid\nMaster in Vitreo-Retina. IOBA. University of Valladolid\nFellow of the European Board of Ophthalmology. Paris\nMaster in Research in Ophthalmology. University of Valladolid\t2003-2009\n2012-2016\n2016-2017\n2016\n2012-2013\n\t\nEmployments (company and positions)\tDate (Year)\nResident in Ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl.\nFellow in Vitreo-Retina. IOBA. University of Valladolid\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. \n\t2012-2016\n2016-2017\n2017-today\n\n2019-Today\n\n\n\t\nClinical Research Experience (tasks and role)\tDate (Year)\nAssociated investigator\n\n' FIS PI20/00740: DESARROLLO DE UNA CALCULADORA DE RIESGO DE\nAPARICION DE RETINOPATIA DIABETICA BASADA EN TECNICAS DE IMAGEN MULTIMODAL EN PACIENTES DIABETICOS TIPO 1. Grant by: Ministerio de Ciencia e Innovacion \n\n' (BIO/VA23/14) Estudio clínico multicéntrico y prospectivo para validar dos\nbiomarcadores ubicados en los genes p53 y MDM2 en la predicción de los resultados funcionales de la cirugía del desprendimiento de retina regmatógeno. Grant by: Gerencia Regional de Salud de la Junta de Castilla y León.\n' Estudio multicéntrico, aleatorizado, con enmascaramiento doble, en 2 grupos\nparalelos y de 52 semanas de duración para comparar la eficacia, seguridad e inmunogenicidad de SOK583A1 respecto a Eylea® en pacientes con degeneración macular neovascular asociada a la edad' (CSOK583A12301; N.EUDRA: 2019-004838-41; FASE III). Grant by Hexal AG\n\n' Estudio de fase III, aleatorizado, doble ciego, con grupos paralelos, multicéntrico para comparar la eficacia y la seguridad de QL1205 frente a Lucentis® en pacientes con degeneración macular neovascular asociada a la edad. (EUDRACT: 2018-004486-13). Grant by Qilu Pharmaceutical Co\n\n' Estudio NEUTON: Ensayo clinico en fase IV para evaluar la eficacia de aflibercept en pacientes Naive con Edema MacUlar secundario a Oclusion de Vena CenTral de la Retina (OVCR) en regimen de tratamientO iNdividualizado Treat and Extend (TAE)”, (2014-000975-21). Grant by Fundacion Retinaplus\n\n' Evaluación de la seguridad y bioactividad de anillos de tensión capsular en conejo. Proyecto Procusens. Grant by AJL, S.A.\n\n'Estudio epidemiológico, prospectivo, multicéntrico y abierto\\npara valorar la frecuencia de la conjuntivitis adenovírica diagnosticada mediante el test AdenoPlus®\\nTest en pacientes enfermos de conjuntivitis aguda”\\n. National, multicenter study. Grant by: NICOX.\n\nEuropean multicentric trial: 'Evaluation of clinical outcomes following the use of Systane Hydration in patients with dry eye”. Study Phase 4. Grant by: Alcon Labs'\n\nVLPs Injection and Activation in a Rabbit Model of Uveal Melanoma. Grant by Aura Bioscience\n\nUpdating and characterization of a rabbit model of uveal melanoma. Grant by Aura Bioscience\n\nEnsayo clínico en fase IV para evaluar las variantes genéticas de la vía del VEGF como biomarcadores de eficacia del tratamiento con aflibercept en pacientes con degeneración macular asociada a la edad (DMAE) neovascular. Estudio BIOIMAGE. IMO-AFLI-2013-01\n\nEstudio In-Eye:Ensayo clínico en fase IV, abierto, aleatorizado, de 2 brazos,\nmulticçentrico y de 12 meses de duración, para evaluar la eficacia y seguridad de un régimen de PRN flexible individualizado de 'esperar y extender' versus un régimen PRN según criterios de estabilización mediante evaluaciones mensuales de inyecciones intravítreas de ranibizumab 0,5 mg en pacientes naive con neovascularización coriodea secunaria a la degeneración macular relacionada con la edad. CP: CRFB002AES03T\n\nTREND: Estudio Fase IIIb multicéntrico, randomizado, de 12 meses de\nseguimiento con evaluador de la agudeza visual enmascarado, para evaluar la eficacia y la seguridad de ranibizumab 0.5mg en un régimen de tratar y extender comparado con un régimen mensual, en pacientes con degeneración macular neovascular asociada a la edad. CP: CRFB002A2411 Código Eudra CT:\n2013-002626-23\n\n\n\nPublications\t\n\n2021\n\n\n\n\n2015\n\n\n\n\n2021\n\n\n\n\n\n2021\n\n\n\n\n2015\n\n\n\n\n2015\n\n\n2014\n\n\n\n\n2015-16\n\n\n\n2015\n\n\n2014\n\n\n2014\n\n\n\n\n2014\n\n\n\n\n\n\n\n2014\n\nJose Carlos Pastor; Jimena Rojas; Salvador Pastor-Idoate; Salvatore Di Lauro; Lucia Gonzalez-Buendia; Santiago Delgado-Tirado. Proliferative vitreoretinopathy: A new concept of disease pathogenesis and practical\nconsequences. Progress in Retinal and Eye Research. 51, pp. 125 - 155. 03/2016. DOI: 10.1016/j.preteyeres.2015.07.005\n\n\nLabrador-Velandia S; Alonso-Alonso ML; Di Lauro S; García-Gutierrez MT; Srivastava GK; Pastor JC; Fernandez-Bueno I. Mesenchymal stem cells provide paracrine neuroprotective resources that delay degeneration of co-cultured organotypic neuroretinal cultures.Experimental Eye Research. 185, 17/05/2019. DOI: 10.1016/j.exer.2019.05.011\n\nSalvatore Di Lauro; Maria Teresa Garcia Gutierrez; Ivan Fernandez Bueno. Quantification of pigment epithelium-derived factor (PEDF) in an ex vivo coculture of retinal pigment epithelium cells and neuroretina.\nJournal of Allbiosolution. 2019. ISSN 2605-3535\n\nSonia Labrador Velandia; Salvatore Di Lauro; Alonso-Alonso ML; Tabera Bartolomé S; Srivastava GK; Pastor JC; Fernandez-Bueno I. Biocompatibility of intravitreal injection of human mesenchymal stem cells in immunocompetent rabbits. Graefe's archive for clinical and experimental ophthalmology. 256 - 1, pp. 125 - 134. 01/2018. DOI: 10.1007/s00417-017-3842-3\n\n\nSalvatore Di Lauro, David Rodriguez-Crespo, Manuel J Gayoso, Maria T Garcia-Gutierrez, J Carlos Pastor, Girish K Srivastava, Ivan Fernandez-Bueno. A novel coculture model of porcine central neuroretina explants and retinal pigment epithelium cells. Molecular Vision. 2016 - 22, pp. 243 - 253. 01/2016.\n\nSalvatore Di Lauro. Classifications for Proliferative Vitreoretinopathy ({PVR}): An Analysis of Their Use in Publications over the Last 15 Years. Journal of Ophthalmology. 2016, pp. 1 - 6. 01/2016. DOI: 10.1155/2016/7807596\n\nSalvatore Di Lauro; Rosa Maria Coco; Rosa Maria Sanabria; Enrique Rodriguez de la Rua; Jose Carlos Pastor. Loss of Visual Acuity after Successful Surgery for Macula-On Rhegmatogenous Retinal Detachment in a Prospective Multicentre Study. Journal of Ophthalmology. 2015:821864, 2015. DOI: 10.1155/2015/821864\n\nIvan Fernandez-Bueno; Salvatore Di Lauro; Ivan Alvarez; Jose Carlos Lopez; Maria Teresa Garcia-Gutierrez; Itziar Fernandez; Eva Larra; Jose Carlos Pastor. Safety and Biocompatibility of a New High-Density Polyethylene-Based\nSpherical Integrated Porous Orbital Implant: An Experimental Study in Rabbits. Journal of Ophthalmology. 2015:904096, 2015. DOI: 10.1155/2015/904096\n\nPastor JC; Pastor-Idoate S; Rodríguez-Hernandez I; Rojas J; Fernandez I; Gonzalez-Buendia L; Di Lauro S; Gonzalez-Sarmiento R. Genetics of PVR and RD. Ophthalmologica. 232 - Suppl 1, pp. 28 - 29. 2014\n\nRodriguez-Crespo D; Di Lauro S; Singh AK; Garcia-Gutierrez MT; Garrosa M; Pastor JC; Fernandez-Bueno I; Srivastava GK. Triple-layered mixed co-culture model of RPE cells with neuroretina for evaluating the neuroprotective effects of adipose-MSCs. Cell Tissue Res. 358 - 3, pp. 705 - 716. 2014.\nDOI: 10.1007/s00441-014-1987-5\n\nCarlo De Werra; Salvatore Condurro; Salvatore Tramontano; Mario Perone; Ivana Donzelli; Salvatore Di Lauro; Massimo Di Giuseppe; Rosa Di Micco; Annalisa Pascariello; Antonio Pastore; Giorgio Diamantis; Giuseppe Galloro. Hydatid disease of the liver: thirty years of surgical experience.Chirurgia italiana. 59 - 5, pp. 611 - 636.\n(Italia): 2007. ISSN 0009-4773\n\nChapters in books\n\t\n' Salvador Pastor Idoate; Salvatore Di Lauro; Jose Carlos Pastor Jimeno. PVR: Pathogenesis, Histopathology and Classification. Proliferative Vitreoretinopathy with Small Gauge Vitrectomy. Springer, 2018. ISBN 978-3-319-78445-8\nDOI: 10.1007/978-3-319-78446-5_2. \n\n' Salvatore Di Lauro; Maria Isabel Lopez Galvez. Quistes vítreos en una mujer joven. Problemas diagnósticos en patología retinocoroidea. Sociedad Española de Retina-Vitreo. 2018.\n\n' Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor Jimeno. iOCT in PVR management. OCT Applications in Opthalmology. pp. 1 - 8. INTECH, 2018. DOI: 10.5772/intechopen.78774.\n\n' Rosa Coco Martin; Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor. amponadores, manipuladores y tinciones en la cirugía del traumatismo ocular.Trauma Ocular. Ponencia de la SEO 2018..\n\n' LOPEZ GALVEZ; DI LAURO; CRESPO. OCT angiografia y complicaciones retinianas de la diabetes. PONENCIA SEO 2021, CAPITULO 20. (España): 2021.\n\n' Múltiples desprendimientos neurosensoriales bilaterales en paciente joven. Enfermedades Degenerativas De Retina Y Coroides. SERV 04/2016. \n' González-Buendía L; Di Lauro S; Pastor-Idoate S; Pastor Jimeno JC. Vitreorretinopatía proliferante (VRP) e inflamación: LA INFLAMACIÓN in «INMUNOMODULADORES Y ANTIINFLAMATORIOS: MÁS ALLÁ DE LOS CORTICOIDES. RELACION DE PONENCIAS DE LA SOCIEDAD ESPAÑOLA DE OFTALMOLOGIA. 10/2014.",institutionString:null,institution:null},{id:"243698",title:"Dr.",name:"Xiaogang",middleName:null,surname:"Wang",slug:"xiaogang-wang",fullName:"Xiaogang Wang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/243698/images/system/243698.png",biography:"Dr. Xiaogang Wang, a faculty member of Shanxi Eye Hospital specializing in the treatment of cataract and retinal disease and a tutor for postgraduate students of Shanxi Medical University, worked in the COOL Lab as an international visiting scholar under the supervision of Dr. David Huang and Yali Jia from October 2012 through November 2013. Dr. Wang earned an MD from Shanxi Medical University and a Ph.D. from Shanghai Jiao Tong University. Dr. Wang was awarded two research project grants focused on multimodal optical coherence tomography imaging and deep learning in cataract and retinal disease, from the National Natural Science Foundation of China. He has published around 30 peer-reviewed journal papers and four book chapters and co-edited one book.",institutionString:null,institution:null},{id:"7227",title:"Dr.",name:"Hiroaki",middleName:null,surname:"Matsui",slug:"hiroaki-matsui",fullName:"Hiroaki Matsui",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Tokyo",country:{name:"Japan"}}},{id:"312999",title:"Dr.",name:"Bernard O.",middleName:null,surname:"Asimeng",slug:"bernard-o.-asimeng",fullName:"Bernard O. Asimeng",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Ghana",country:{name:"Ghana"}}},{id:"318905",title:"Prof.",name:"Elvis",middleName:"Kwason",surname:"Tiburu",slug:"elvis-tiburu",fullName:"Elvis Tiburu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Ghana",country:{name:"Ghana"}}},{id:"336193",title:"Dr.",name:"Abdullah",middleName:null,surname:"Alamoudi",slug:"abdullah-alamoudi",fullName:"Abdullah Alamoudi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"318657",title:"MSc.",name:"Isabell",middleName:null,surname:"Steuding",slug:"isabell-steuding",fullName:"Isabell Steuding",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}},{id:"318656",title:"BSc.",name:"Peter",middleName:null,surname:"Kußmann",slug:"peter-kussmann",fullName:"Peter Kußmann",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}}]}},subseries:{item:{id:"2",type:"subseries",title:"Prosthodontics and Implant Dentistry",keywords:"Osseointegration, Hard Tissue, Peri-implant Soft Tissue, Restorative Materials, Prosthesis Design, Prosthesis, Patient Satisfaction, Rehabilitation",scope:"