Planned versus actual coverage of the survey.
\r\n\tCongenital hearing loss means hearing loss that is present at birth. I have managed children with hearing loss for many years, and the most touching thing is the light that blooms on the face while the hearing-impaired child heard his mother's voice at first time. The scene of "happy tears" impressed me so much. To hear the voice that has not been heard is so pleasant, as if this ordinary listening experience is a supreme listening enjoyment.
\r\n\r\n\tAge-related hearing loss means a progressive loss of ability to hear high frequencies with aging, also known as presbycusis. Among them are the influence of internal and external factors such as genes, drugs and noise exposure. The studies pointed out that the brain stimulation of the hearing-impaired person is greatly reduced compared with subjects with normal hearing. The connection of auditory cortex and other brain areas has declined a lot, which is probably one of the important causes of dementia or even depression in the elderly.
\r\n\r\n\tNoise-induced hearing loss is hearing impairment resulting from exposure to loud sound. There is actually continuous and endless noise in many workplaces, which may cause chronic and cumulative damage. Some young people often work hard but easily neglect to protect themselves. In addition, in recent years, entertainment noise (such as nightclubs, concerts, and personal listening devices) has caused hearing impairment in young people. These should be avoidable and preventable.
\r\n\r\n\tHearing Science is the study of impaired auditory perception, the technologies and other rehabilitation strategies for persons with hearing loss. Public health has been defined as "the science and art of preventing disease", improving quality of life through organized efforts. To avoid the “epidemic” of hearing loss, it is necessary to promote early screening, use hearing protection, and change public attitudes toward noise.
\r\n\r\n\tBased on these concepts, the book incorporates updated developments as well as future perspectives in the ever-expanding field of hearing loss. Besides, it is also a great reference for audiologists, otolaryngologists, neurologists, specialists in public health, basic and clinical researchers.
",isbn:"978-1-83968-678-8",printIsbn:"978-1-83968-677-1",pdfIsbn:"978-1-83968-679-5",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"a4b7dbb02ba00e7412422cd5dbffa029",bookSignature:"Dr. Tang-Chuan Wang",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10529.jpg",keywords:"Hidden Hearing Loss, Plasticity, Electrophysiology, Otoacoustic Emission, Newborn Hearing Screening, Genetics, Aging, Hearing Aids, Noise Exposure, Occupational Hearing Loss, Epidemiology, Prevention",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"September 3rd 2020",dateEndSecondStepPublish:"October 1st 2020",dateEndThirdStepPublish:"November 30th 2020",dateEndFourthStepPublish:"February 18th 2021",dateEndFifthStepPublish:"April 19th 2021",remainingDaysToSecondStep:"3 months",secondStepPassed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dr. Tang-Chuan Wang is an excellent otolaryngologist-head and neck surgeon in Taiwan; a research scholar of Harvard Medical School and University of Iowa Hospitals. He worked in the Hospital of the University of Pennsylvania, Boston Children's Hospital, and Massachusetts Eye and Ear. Due to his contribution to biomedical engineering, he was invited into the executive committee of HIWIN-CMU Joint R & D Center in Taiwan.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"201262",title:"Dr.",name:"Tang-Chuan",middleName:null,surname:"Wang",slug:"tang-chuan-wang",fullName:"Tang-Chuan Wang",profilePictureURL:"https://mts.intechopen.com/storage/users/201262/images/system/201262.gif",biography:'Dr. Tang-Chuan Wang is an excellent otolaryngologist – head and neck surgeon in Taiwan. He is also a research scholar of Harvard Medical School and University of Iowa Hospitals. During his substantial experience, he worked in Hospital of the University of Pennsylvania, Boston Children\'s Hospital and Massachusetts Eye and Ear. Besides, he is not only working hard on clinical & basic medicine but also launching out into public health in Taiwan. In recent years, he devotes himself to innovation. He always says that "in theoretical or practical aspects, no innovation is a step backward". 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From chapter submission and review, to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"50619",title:"Rejuvenation on the Road to Pluripotency",doi:"10.5772/63219",slug:"rejuvenation-on-the-road-to-pluripotency",body:'\nIn their recent Overview on Chronic Disease, the Center for Disease and Control (CDC) cited multiple studies in the last few years highlighting the predominance (86% in 2010) of chronic diseases in the US national health care spending. Each of these diseases has their own treatments directed by their individualized fields of research. Yet the key risk factor and fundamental correlations behind many conditions like arthritis ($128 billion in 2003), heart disease and stroke ($315.4 billion in 2010), and type II diabetes ($245 billion in 2012) are of course middle and advanced age [1]. To further quantify the core role of aging, a 2013 study did a prospective study and estimated that the economic value of delaying aging in Americans by only about 2 years would save $7.1 trillion over 50 years [2]. This study was prompted by the growing field of aging research and longevity technologies, but now an even bolder direction is emerging. Researchers are venturing beyond just slowing age or promoting healthy aging and actually attempting to reverse the manifestations of age, from the cellular to the tissue and organ levels.
\nIn another field of research, stem cell science has rapidly grown in the last few decades as a means of studying tissue development and maintenance and to develop methods to artificially produce cells that are either absent or dysfunctional in patients. Again the applications here are pathology specific, conditions like blood and immunological diseases, brain and spinal cord injury, and type I diabetes, as well as some of the more aging‐correlated disease like Alzheimer\'s and heart disease [3]. A unifying approach in the field is the use of pluripotent stem cells, the crown jewel state that can produce any cell in the body. Natural and artificial processes all reach this state by starting with differentiated cell types and “reprogramming” them back to pluripotency. Clearly, this process of reprogramming represents a drastic change and powerful technology, whose potential is just beginning to be explored.
\nThe focus of this chapter is convergence of these two fields. We will see how rejuvenation is intimately linked to reprogramming. Furthermore, we discuss how technologies that induce pluripotency may hold the key to a wholescale and stable reset of cellular age, with tissue‐ and organ‐level consequences.
\nRejuvenation is actually not at all an esoteric idea. It happens consistently and is crucial to the survival of virtually every species. Most would immediately think of relatively simple organisms like the hydra and the jellyfish Turritopsis dohrnii, which have virtually unlimited regenerative potential [4]. Yet there is a rejuvenation mechanism in much more “complicated” animals, and most importantly in humans, which hides in plain sight: the process of reproduction. The most popular and widely investigated feature of this process is attaining pluripotency, the cellular state that can differentiate into the full diversity of cells in a new organism, starting from two highly specialized and differentiated cells (i.e., the germ cells). However, rejuvenation is an equally exciting phenomenon that occurs during reproduction and is only beginning to be explored. A simple input output analysis verifies this: The inputs are the sperm and oocyte cells of aged parents, typically in their second or third decade of life, while the outputs are the myriad of different cells that make up a new, young organism, retaining none of aged aspects of the parent cells. While one could argue that germ cells have evolved special mechanisms that, unlike somatic cells, prevent them from aging, multiple studies have shown that germ cells do indeed age. Sperm show progressively accumulated DNA damage, elevated reactive ion species levels, and loss of chromatin integrity with age [5]. Oocytes show altered expression of genes implicated in DNA methylation and histone acetylation and mitochondrial dysfunction with age, as well as microniche‐imposed aging, through nutrition and hormonal pathways [6]. If these manifestations of age are not erased during reproduction, each generation would progressively accumulate age and the species would no longer have the capability to produce viable offspring after just a few generations.
\nThe mechanisms specifically behind the reversal of age during reproduction are largely unknown. However, there are two main categories of changes that drive embryogenesis as a whole: genetic and epigenetic. In terms of genetic, the main change is the genetic recombination of the parental chromosomes, which are gathered during zygote formation, right after fertilization, and fused during the first mitotic division into blastomeres [7]. The result is a new genome derived from the parental genomes. There are also more active DNA repair process but they can be tied to upregulation of genes for double‐strand break repair and cell cycle checkpoint control that result from the epigenetic changes we will cover next [8]. The reactivated repair processes would definitely have an age‐reversing effect, but there is no evidence that the recombination to produce a new genome, alone, should rejuvenate the cell. Epigenetics also undergo large shifts in the initial stage of zygote formation, characterized by a genome‐wide loss of DNA methylation except in certain centromeric and parentally imprinted regions and retrovirally derived repetitive elements. In addition, there are widespread alterations to methylation and acetylation of H3 and H4 histones from the maternal genome, while the paternal genome abandons its own protamines for these modified histones as well. Increased activity of methlytransferase has also been observed during this stage; these types of enzymes preserve methylation patterns during cell division for both DNA and histones. Together, these changes further regulate overall heterochromatin organization [9]. Subsequent stages in pre‐implantation development up until blastocyst formation show a more passive loss of DNA methylation with cell division, primarily due to reduced levels of methlytransferase in the cell nucleus. The blastocyst then marks the beginning of lineage specification, with an increase in methylation and additional histone modifications that are primarily correlated to differentiation rather than rejuvenation [10]. Ultimately, the bulk erasure of parental epigenetic markers, for cell type and most likely for age, is during the reprogramming of the parental germ cells into a totipotent zygote; totipotency here refers to the zygote\'s ability to differentiate into all embryonic and extraembryonic (supporting) tissue.
\nArtificial reprogramming to totipotency was further pursued in the last few decades, with the primary objective of attaining cells that could differentiate into any desired cell type. The first key deviation from the natural process was the technology of somatic cell nuclear transfer (SCNT). In fertilization, only the sperm\'s nuclei is transferred and it is the oocyte that provides the reprogramming environment. SCNT replaces the sperm nucleus with a somatic cell nucleus for fertilization, but the overall reprogramming process is fundamentally the same, again with large‐scale genetic and epigenetic shifts to yield a viable, young offspring. [11] This method, however, established that somatic cells, in addition to germ cells, could be reprogrammed, and thus rejuvenated. The next key breakthrough, which is our main focus, is the technology of induced pluripotent stem cell (iPSC) reprogramming. This technology focuses on reprogramming somatic cells to pluripotent cells, which can specifically form only the embryonic tissues and thus is more relevant than totipotency. More importantly, the strategy here is substantially different. Instead of the reprogramming driven by the oocyte, iPSC technologies introduce exogenous pluripotency genes into somatic cells to induce reprogramming to pluripotency [12]. This process does not include a genetic recombination of two different genomes, like reproduction or SCNT. Thus, the success of this technology established that a global shift in the gene expression profile, induced by epigenetic remodeling, without substantial genetic alterations is sufficient to execute reprogramming and, as we will see in the next section, can also implement multiple changes toward rejuvenation.
\niPSC reprogramming was developed to artificially attain the embryonic stem cell (ESC) state, a derivative of the totipotent zygote state established by natural fertilization and SCNT reprogramming. The principle behind this technology is that forced overexpression of genes that normally maintain the pluripotent state in ESCs is sufficient to induce reprogramming in somatic cells to revert back to an ESC‐like state. The most widely utilized strategy, which most of the result we discuss have applied, is the exogenous introduction of the genes Oct4, Sox2, Klf4, and cMyc, typically achieved by viral or episomal vectors [12]. In evaluating the final product of these procedures, it has been noted that the transition to pluripotency is accompanied by multiple indications of rejuvenation that, in most cases, are retained upon subsequent redifferentiation of the iPSC. As iPSC reprogramming is a cellular technology, it requires a set of cellular biomarkers by which to measure age. The search is still ongoing for a comprehensive and precise list, but the “Hallmarks of Aging” presented by Lopez‐Otin et al. in 2013 provide one most generally accepted categorization of the known biomarkers [13]. We will review the multiple signs of rejuvenation through iPSC reprogramming in the context of these hallmarks.
\nEpigenetics include all the changes to the structure of DNA, but not the DNA sequence itself, which control transcriptional potential and ultimately gene expression. Epigenetics most notably establishes cell type and functionality, but also includes many markers of age. DNA methylation is a key aspect of epigenetics and aging correlates with a global trend toward hypomethlyation in numerous cells types across tissues [14]. This is linked to the decline in mRNA transcription for DNA methyltransferase (DNMT), which controls the transfer of methylation patters to daughter cells [15]. Histones, which act as the “spools” around which DNA strands are wound, are another aspect of epigenetics which show age‐related modifications, for instance, trimethlyation levels increase for H3K27 and H3K9 but decrease for H4K20, which are all involved in heterochromatin formation, the most densely packed form of chromatin [16]. A key driver of these alterations to histone methylations is the decreased transcription of methyltransferase proteins with age, like those of the Polycomb and trithorax groups [17]. There is an overall loss of heterochromatin with age, driven by the histone modifications but also lost during cell division itself, as recondensation processes are not perfectly efficient. These global trends lead to a loss in gene silencing with age and overall increase in transcriptional noise. iPSC reprogramming reverses of many of these changes, with a youthful restoration of the levels of HP1γ (a key marker for heterochomatin) and trimethlyated H3K9, increased heterochromatin, but there is still much yet to be explored [18]. Changes in the levels of endogenous Polycomb proteins have not been studied, but the artificial knockdown or their genes yields decreased reprogramming efficiency—hinting at a correlation between the two [19]. There is also an overall decrease in DNA methylation but this is primarily because global hypomethlyation is a characteristic of the pluripotent state, as seen in ESCs. A better comparison, that has yet to be investigated, is between the methylation levels of the original source cells and iPSC‐derived cells of the same cell type. A correlation between artificially increased DNMT and reprogramming efficiency has also been made, but no further study has been done on whether reprogramming itself stimulates DNMT transcription [20]. Ultimately, reprogramming itself is characterized by a large‐scale epigenetic remodeling, most infamously for dedifferentiation but also for rejuvenation.
\nDirect damage to DNA from natural metabolic activities, stochastic chemical (endogenous and environmental), and radiative interactions regularly occurs throughout life at rate of 105 total molecular lesions per cell per day [21]. There are number of enzymatic repair mechanisms in place to fix these aberrations, but these processes are not perfectly efficient and some processes, like homologous recombination (HR) and non‐homologous end joining (NHEJ), which occur during cell division, also decline in efficiency with age [22, 23]. This leads to the accumulated DNA damage, which impairs cell functionality, and decreases the number of viable progeny for proliferative cells; this is the central idea in the DNA damage theory of aging that leads to tissue wide consequences in organs like brain, liver muscle, and kidney. In addition, the nuclear lamina degrades with age with conformational defects like folding and blebbing; this impairs the lamina\'s function as the overall structural support for the nuclear material, especially heterochromatin in the epigenetics hallmark. These nuclear lamina defects result from age‐related decreases in production of laminar proteins, like lamina‐associated protein 2α (LAP2α), and altered distribution of these proteins, like the localization of the laminar matrix protein lamin A/C to the nuclear rim but not the nucleoplasm [24]. iPSCs show evidence of reducing accumulated DNA damage through a marked upregulation in genes and activity in both the HR and NHEJ pathways. Further evidence for reinvigorated repair mechanism are the restored levels of phosphorylated histone H2AX which is a key biomarker for the repair of double‐strand breaks, in the iPSC state and after redifferentiation. In addition, lamina structure is shown to be restored in the iPSC state which replenished levels of LAP2α. In general, reprogrammed cells also show as an increased sensitivity to extreme irradiation damage noted by a greater propensity to undergo apoptosis, to eliminate mutated cells; this is ultimately healthier for the tissue by clearing cells that may instigate cancerous growth [25].
\nDuring cell division, helicase “unzips” DNA in one direction and DNA polymerases must build off of the two template strands but can only move in the 5′→3′ direction. Thus, one polymerase can only move oppositely to the helicase, meaning it must repeatedly attach, synthesize, detach, and catch up to the helicase. Attachment sites are directed by RNA primers, which bind to DNA behind the helicase. Synthesis can only start after the primer, so the DNA by covered primer is not synthesized until the subsequent synthesis when it detaches and the polymerase starts further upstream. The problem arises at the end of the chromosome, where it is no further upstream site for polymerase to begin at and thus the end is simply not copied. Nature\'s solution is to pad the end of the chromosomes with telomeres, repeats of noncoding junk DNA (TTAGGG), which are lost instead of coding segments. Telomere erosion is characteristic of cell division and thus aging itself. A number of studies have shown an elongation of telomeres, around 40%, upon reprogramming to iPSC [26–28]. This elongation of telomeres is mechanistically tied to the reactivated transcription of telomerase, a reverse transcriptase enzyme which synthesizes additional telomere repeats. Genes for telomerase are only expressed in stem cells as well as some other highly proliferative somatic cells, but they are silenced upon differentiation. Upon redifferentiation, all lines have been shown to have shorter telomeres than the original iPSC but some who have shown still have longer telomeres than the original source cell [27, 28]. This subsequent loss of telomere length is not necessarily due to accelerated telomere attrition, but just due to the lengthy redifferentiation protocols and variations in each cell line\'s reprogramming and redifferentiation efficiency. If these protocols can be made shorter and more efficient, the rejuvenation effects could be better retained. This is another example of how a genetic hallmark is addressed by alterations in gene expression. It has also yet to be assessed exactly when telomerase is reactivated and when it is lost during redifferentiation; this knowledge may help to further optimize the retention of this rejuvenation effect.
\nOlder cells exhibit a greater percentage of mitochondria in the less efficient “condensed” and “ultra‐condensed” configurations than in the healthier “orthodox” configuration; the former have their inner and outer mitochondrial membranes further apart, promoting a smaller matrix and larger cristae. This condensed structure leads to a lower transmembrane potential, which is correlated with higher, less regulated respiratory activity. iPSC\'s and their redifferentiated lineages exhibit a reversal of this trend, with a higher proportion of orthodox mitochondria as well as higher membrane potentials when compared with the aged source cells. In addition, reprogramming and redifferentiation studies have also shown decreased mitochondrial mass, increased levels of adenosine triphosphate (ATP), and decreased reactive oxygen species (ROS) levels, which are all characteristic of a more youthful phenotype [29]. The lowered ROS levels are especially promising, as elevated levels are a key player in the free radical theory of aging. This idea holds that the electrically active by‐products of metabolic reactions, like ROS, can aberrantly ionize the biomolecular machinery of cells, ultimately impairing the functionality of mitochondria and other cell compartments. These youthful changes can be fundamentally tied to upregulated mitochondrial biogenesis genes like PGC‐1 alpha and antioxidant genes like GPX1 during reprogramming [30]. This upregulated biogenesis may directly explain the noted increase in youthful mitochondrial morphology and functionality, while the antioxidant genes explain the loss of ROS and further damage to mitochondria. The time course of these changes during reprogramming has not been documented, with the exception of mitochondrial mass, which has been noted to decrease as early as 3 days after the induction of reprogramming [31]. Interestingly, during SCNT reprogramming, cells began exhibiting higher membrane potential in the first week of reprogramming and were linked to increased expression of Glut1, Pfkm, Hxk2, and Ldha which drive glycolytic activity over oxidative phosphorylation [32]. This transition would explain the decreased ROS levels despite the observed increase in ATP, as glycolysis also produces ATP but without the production of ROS. This initial week, however, does not show the onset of pluripotency, as marked by the expression of pluripotency genes like Nanog and Oct4, suggesting a temporal segregation of the gene expression changes driving rejuvenation from those that drive differentiation.
\nProteostasis is regulated by modulating levels of both protein transcription/stabilization, to ensure the cell has the proteins it needs, and proteolytic activity, to prevent proteotoxicity. The former, of course, is a direct result of gene expression dynamics. The latter is implemented through the ubiquitin‐proteasome and autophagy‐lysosome systems; the activity of both notably diminishes with age. This leads to an aggregation of macromolecules as well as degraded organelles that contribute to other aging hallmarks—like mitochondrial dysfunction and senescence [32, 34]. Reprogramming studies have explored relatively little in regard to rejuvenated proteolytic activity. The key finding so far has been an elevated level of autophagy upon induction of reprogramming, peaking on day 2 then relaxing to basal levels from day 3 on until pluripotency is reached [35, 36]. This transient activity has been linked to increased mitophagy, a key role of autophagy, which is responsible for the clearing of degraded mitochondria. This works in tandem with the aforementioned increased mitochondrial biogenesis to create the noted skew in the mitochondria distribution to the younger, orthodox configuration [26]. It also postulated that the additional autophagy is responsible for eliminating cell type‐specific protein complexes early on, thus promoting dedifferentiation. This effect may be transient, but the additional degraded proteins and organelles that are removed during the interim definitely contribute to cell rejuvenation. It is crucial to note that these studies were on murine cells, so translation of these effects to human systems also needs to be investigated. Proteasomal activity has yet to be studied in the context of iPSC reprogramming, but increased activities have been noted in human ESCs, linked to elevated expression of the PSMD11 gene [38].
\nSenescence is defined as the state of full mitotic arrest, relevant primarily for proliferative cells. In vitro, this is linked to the Hayflick limit—the point at which cells have undergone enough cell divisions to deplete their telomeres. In vivo, senescence is also linked to the accumulated DNA damage with age and triggered by epigenetic (chromatin) remodeling at the INK4a/ARF locus. Senescence is likely a programmed biological mechanism for preventing the propagation of cells with damaged DNA, which is more prevalent in aged cells as characterized by shortened telomeres. This prevents mutation‐induced cancerous growth and aberrant functionality, but it also arrests tissue growth and repair in general. iPSC reprogramming has been achieved on completely senescent populations, propagated for 51 population doublings then maintained for 2 months, and show revived proliferation in the pluripotent state as well as upon redifferentiation [39]. In addition, expressions of cell cycle‐inhibiting proteins like p16 and p21, which initiate the INK4a/ARF remodeling, are known to increase with age but diminish after reprogramming and redifferentiation. Crucially, the redifferntiated progeny can once again be passaged into senescence, thus verifying that no cancerous growth was induced during the entire process.
\nThis hallmark concerns poorer regulation in gene expression pathways that regulated cell metabolism based on the availability of nutrients. The mTOR family of genes is a key glucose‐ and amino acid‐sensing pathway, while the AMPK pathway is route triggered by adenosine monophosphate (AMP) levels, a by‐product of ATP synthesis. Artificial downregulation of mTOR and upregulation of AMPK genes have both led to increased longevity in model organisms, while the inverse (as occurs naturally) has been linked with many age‐related pathologies like diabetes, hinting at epigenetic alterations to these pathways with age [40, 41]. During reprogramming, mTOR was downregulated in the same pattern that autophagy increased and AMPK activity was upregulated in the first 6 days; both pathways are drivers for autophagy [31, 36]. In addition, downregulation of mTOR is also involved in mitochondrial biogenesis [36]. Thus, the deregulated nutrient‐sensing hallmark further links the loss of proteostasis and mitochondrial dysfunction hallmarks. The two remaining FOXO and Sirtuin pathway have yet to be studied for their role in reprogramming.
\nThere are two remaining hallmarks. The hallmark of stem cell exhaustion is defined by age‐associated losses in both proliferative capacity and stem cells reserves, like in hippocampal neural stem cell populations, as well as altered epigenetics for differentiation potential, for instance, aged hematopoietic stem cells bias toward myeloid lineages over lymphoid lineages [42, 43]. The hallmark of altered intracellular communication results from increased expression and secretion of pro‐inflammatory cytokines by senescent and pre‐senescent cells as well as proteotoxicity from the loss of proteostasis. This leads to a larger, immune system phenotype of “inflamaging,” an age‐induced propensity to sustain a chronic low‐grade innate immune response in multiple tissues, which ultimately interferes with single cell function as well as cell‐to‐cell interactions [44]. To our knowledge, no iPSC reprogramming studies have really examined rejuvenation of these hallmarks, primarily because both truly need be assessed within aged tissue. The organoid technology we will discuss in the next section may offer the platform to evaluate the rejuvenation of these hallmarks as well through iPSC reprogramming.
\nThese rejuvenated biomarkers are summarized pictorially in Figure 1, but there are still many other known biomarkers within these hallmarks that must be assessed for iPSC‐ and iPSC‐derived cells and, as new ones are being discovered, they must also be evaluated. In addition, the results are primarily in fibroblasts, the most common source material for iPSC reprogramming, so the generalization of the rejuvenation effect to other cell types still needs to be assessed. Still, these studies provide initial evidence that iPSC reprogramming can reset age to some degree. In the next section, we will see where this technology stands as a possible rejuvenation procedure.
\nCellular Hallmarks of aging and rejuvenation through iPSC reprogramming.
The field of iPSC reprogramming was developed to attain and harness the power of pluripotency. The rejuvenation effects that accompany the process were secondary discovery, though one with great potential as well. As we now move to capitalize on this effect, we must evaluate it in the broader field of aging research and technologies. In this section, we will first explore how the mechanistic principle behind this iPSC rejuvenation fits in with unifying theories of aging. Then we will assess the reprogramming strategy in the context of other technologies for healthy aging and rejuvenation. Finally, we will discuss the future perspectives and outlook of this technology in terms of scalability and clinical application.
\nThe common theme we note from all the hallmarks are that multifaceted changes in gene expression that are linked to and propagate the aging phenotype. We similarly saw that the rejuvenation effects of iPSC reprogramming could be traced to alterations in gene expression. We noted a few examples, yet these are but a small sample of the full gamut of changes that drive rejuvenation along with dedifferentiation. The core driver of these changes in iPSC reprogramming is the massive overhaul of the epigenetic landscape, which as previously noted, alters the transcription potential throughout the genome. Additionally, the noted changes in nuclear lamina influence epigenetics and further regulate the distribution of transcription factors as well as the transport of mRNA for translation. Furthest downstream, the increased proteolytic activity, though transient, also clears out gene products—another form of gene regulation. The key concept this strategy has tapped into is the so‐called programmed aging hypothesis [45]. This idea holds that the aging phenotype is driven in large part by deterministic and programmed changes, and thus is reversible. At the cellular level, this directly points to changes in gene expression, which is reversible especially through the modes of epigenetic and nuclear lamina modifications. So if age is programmed, iPSC technology could possibly reprogram age—truly apt naming in hindsight. The dual theory is the damage‐induced aging hypothesis, which holds that aging is driven by the stochastic degradation of multiple cellular components. This damage is driven by environmental interactions as well as internal degradation as a result of metabolic processes [46]. We discussed manifestations of this in previous section, such as DNA damage, ROS damage, and proteotoxicity from accumulated macromolecules. Unlike programmed aging, the results of this damage are random and thus are not inherently reversible. iPSC reprogramming cannot directly oppose damage but it can help to mitigate some of this damage. As we have seen, it can boost expression for natural repair mechanisms, like homologous repair of DNA damage; it can promote the synthesis of new organelles. From an evolutionary perspective, the two hypotheses may be fundamentally linked. The continued accumulation of age‐related damage and the resulting loss in functionality may make retaining older individual less beneficial from a species‐level perspective. Older individuals would be less fit and less capable of performing their role in a communal society, more susceptible to and may further transmit pathogens, more likely to produce mutated or dysfunctional offspring, and still take up resources that could go to the younger generations. Thus, species may have evolved programmed mechanisms to further the decay with age and thus increase the mortality and clearing of the older individuals. This could explain why a natural rejuvenation exists but only occurs in the production of the next generation, instead of somehow being applied to retain the youthful phenotype [6], like in the case of mitochondrial biogenesis, and it can boost the clearing of damaged components by transiently increasing proteolytic activity. At the same time, there are age‐induced damages that iPSC reprogramming cannot counter (for instance, genetic mutations or damage) without an intact reference template. So in regard to damage‐induced aging, there are definitely inherent limitations to this gene expression level technology.
\nThe predominant strategy in the field of aging is to identify individual gene expression pathways that maintain the youthful phenotype or advance aging and artificially control their expression to promote healthy aging. Treatment involves either controlling the expression of endogenous genes through cues like DNA‐binding agents and small molecules or introducing exogenous genetic material into the cells through vectors (primarily viral and episomal). The strategies have been applied to genes for telomerase, autophagy, mTOR, AMPK, p16, and p21 to name a few of the aging biomarkers we discussed [47]. Additionally, many pharmacological strategies have been developed to induce these pathways with more exogenous control, based on administration and dosage of the drug. Notable examples, specifically in regard to nutrient sensing, include rapamycin, an mTOR inhibitor, metformin, an AMPK activator, and fisetin and resveratrol, both are situin activators [48]. Both the genetic and pharmacologic approaches are fundamentally lacking in two respects. First, these approaches are too narrow in scope. These strategies target one or a handful of genes, but aging is a complex phenotype propagated by a full array of genetic pathways. iPSC reprogramming, in contrast, is more comprehensive with a whole scale epigenetic remodeling that instigates global changes in gene expression which encompass many of the candidate pathways for gene/pharmacological therapies as well as others for aspects like mitochondrial function and gene repair. Even though neither the pathways of aging nor all the gene expression changes in reprogramming are known, the signs of iPSC rejuvenation already discussed and the intuition that iPSC reprogramming mimics natural reprogramming during reproduction both suggest that iPSC reprogramming may offer a more holistic reset of age. The second failing of the traditional genetic/pharmacological approaches is that they are too simplistic in application. These approaches are primarily static over/under expressions of their target genes, but outside of the direct rejuvenation effects, these changes may have detrimental consequences on other cellular processes. For instance, genetic knockout or knockdown of p16 genes may prevent senescence and promote tissue repair, but they increase the chance of spontaneous cancer development [49, 50]. Another example is rapamycin\'s induction of immunosuppressant pathways, which would outweigh its benefit in inhibiting mTOR [51]. A more viable solution would be a dynamic sequence of changes to gene expression which can respond and compensate for each other to produce a stable, youthful homeostasis. Reprogramming also meets this criterion. iPSCs are very similar to the stable, naturally occurring pluripotent state of ESCs, in terms of both epigenetics and gene expression [52]. In addition, redifferentiation of iPSCs yields viable somatic cells which do not show signs of aberrant functionality or advanced aging. In addition, they regain the aged and senescent phenotype after extensive passaging, having avoided cancerous mutations, as noted earlier.
\nAnother direction in the aging field has gained attention in recent years: heterochronic parabiosis, the surgical joining of circulatory system between old and young mice. This promotes more than just longevity or healthy aging; it rejuvenates whole tissue in multiple organs, like muscles, heart, and brain, in the aged mouse. The mechanism behind this is the replacement of old organism\'s aged extracellular milieu—distributed through the bloodstream—with that of a youthful organism [53]. This milieu contains cytokines, hormones, and other signaling molecules which influence and bias gene expression at the single cell level expression through the intracellular communication and nutrient‐sensing hallmarks. Programmed changes to the immune and neuroendocrine systems with age drive the production of this old milieu, and thus this technology also hinges on the programmed aging hypothesis, specifically its extracellular manifestation. The drawback of parabiosis, and immune and hormonal therapies in general, is that the “youthful” milieu is a dynamic and complex mixture, again a homeostatically maintained youthful state. This would require continuous transplantation of youthful blood from another patient, which is not feasible. The stand in would be artificial synthesis and constant administration of the youthful factors in the blood, which is not completely known and would be extremely difficult to develop and maintain. The same is true for immune and hormonal therapies, which also have detrimental side effects; a clear example is the use immunosuppressors would reduce chronic inflamaging but also weaken the response to pathogens. Ultimately, this extracellular rejuvenation approach would require rejuvenation of the entire immune and neuroendocrine systems to regulate and maintain. iPSC reprogramming is still primarily a single cell technology, and an entire rejuvenated system is still far beyond its scope. Yet the technology can still have extracellular effects since every somatic cell secretes signaling molecules that influence the cells in their local microniche. Thus, rejuvenated iPSC‐derived cells, with sufficient numbers, can start to propagate the youthful phenotype in the tissue in which they are transplanted. In addition, multiple groups are working on producing rejuvenated iPSC‐derived lymphoid lineages and T cells. As part of immunoaging, hematopoietic stem cells tend to differentiate more to the myeloid rather than the lymphoid lineage, thus artificial iPSC derivation of the latter can help compensate for this shift [54]. In addition, adaptive immunity to counter specific pathogens diminishes with age due to weakened response by T cells. In vitro production of rejuvenated T cells from iPSCs and transplantation could restore this functionality [55]. The key target that is being explored in iPSC endocrine rejuvenation is the production of insulin producing β‐islet cells, which deplete with age [56]. If iPSC‐derived cells can be used to holistically rejuvenate just these two systems, then it may capture or surpass the effects seen in parabiosis and subsequently rejuvenate the rest of the somatic cells.
\nOutside of single rejuvenated cell production, iPSC technology is showing some progress in developing rudimentary tissue through the use protocols for the derivation of organoids. These protocols are methods of guided differentiation that direct the iPSC to a somatic stem cell fate, instead of a fully differentiated cell. By mimicking the environmental signals during embryo development and providing 3‐D development infrastructure (air‐liquid interface or Matrigel embedding), pluripotent cells can be directed to form a rudimentary organ bud. The cells at this state can be further proliferated or instructed to differentiate into multiple cell types by switching to a differentiation‐promoting media. The final product resembles a fragment of in vivo tissue in its diversity and arrangement of the different cell types; successful examples include intestine, liver, and thyroid [57]. This is still a very nascent technology, with the primary focus of using organoids to study normal and cancerous development. There have not been any studies assessing to what degree the rejuvenation effects of iPSC reprogramming are retained in the final product, especially since there are now two phases of differentiation to reach the initial organ bud and then to attain the fully differentiated tissue. One encouraging sign is that these organoids can be maintained up to and some times longer than 6 months in culture, despite most of the primary cell donors being of advanced age (50s–70s). This has been noted to surpass the Hayflick limit, so at least telomere attrition and senescence may be reversed in the derivation of these organoids. Future studies can investigate the same aging hallmarks in the cells of organoids, but also explore the two additional hallmarks yet untouched in previous studies. They would be able to assess the intercellular communication—how the interactions of different cell types maintain or degrade the youthful phenotype. In addition, the differentiated organoid also retains some somatic stem cells, so these studies can also look at the stem cell exhaustion in maintaining and regenerating the organoid. The organoid platform also provides the opportunity to look at an additional dimension of aging in addressing the rejuvenation of tissue functionality. This moves beyond the Hallmarks of Aging, which are fundamentally at the cellular level, to focus on the emergent function of specific tissues which degrades due to the hallmarks. So far, very little work has been done on assessing organoid functionality. One notable example is a 2015 study that show albumin and bile acid production, ammonia elimination, CYP3A4 activity, and midazolam metabolism in liver organoids, all of which decline with age [58]. These organoids were formed directly from somatic stem cells of the patient, another method for organoid development, and thus show levels of functionality in these areas similar to in vivo. Future studies can use the iPSC derivation and compare this functionality with the in vivo counterpart for a possible rejuvenation effect.
\nEven if iPSC technology progresses to the point where generating whole rejuvenated tissue or organs becomes feasible, transplantation still presents a major hurdle. iPSC‐derived transplant do have the benefit of being autologous, thus avoiding issues like rejection and graft‐vs‐host disease. However, they still require extensive surgical procedures, which would be especially detrimental to aged patients. In addition, heterochronic transplant studies, specifically transplanting muscle from a young organism into an aged one, show that the young tissue succumb to the aged environment within a few months [59]. Multiple aged tissues and organs must be replaced together, possibly even entire systems (like immune and neuroendocrine mentioned earlier), to change the aged environment and get a lasting rejuvenation effect. These obstacles necessitate a procedure that can scale and rejuvenate multiple tissues simultaneously (which even organoids could not eventually do). Ideally, all of this would be in vivo itself. In vivo reprogramming to iPSC has been achieved in mice, using transgenic specimen but also through the injection of exogenous transcription factors. The immediate drawback is that the subsequent differentiation of the iPSCs cannot be guided because they invariably form teratomas. This actually captures the core problem behind both scalability and in vivo use: the undesired dedifferentiation. The technology iPSC reprogramming was developed to replace cells that were depleted or deficient in a patient. The rejuvenation effect of this process was discovered, essentially, as a side effect. Now with the explicit goal of rejuvenation, we can assume that the cells are already present in the patient but they be made younger, so dedifferentiation is unnecessary. This dedifferentiation necessitates complicated, lengthy, and cell type‐specific redifferentiation protocols that can at best generate very rudimentary tissue. They cannot generate multiple tissue types simultaneously, cannot be carried out in vivo, and may erase some of the rejuvenation effects. Reprogramming, itself, is just a series of epigenetic changes so a subset of these must be for dedifferentiation and another subset (possibly overlapping) must be for rejuvenation. The optimal solution, now, would be to identify and execute the epigenetics for rejuvenation without those for dedifferentiation. This may be the best hope for one day utilizing the rejuvenation potential of iPSC reprogramming as a clinical treatment.
\nUltimately, rejuvenation is realized on the road to pluripotency. Harnessing these mechanisms to reset the age of cells in an organism is indeed an audacious goal, but not more audacious than artificially achieving pluripotency itself. One can argue that the true scientific leap was developing the iPSC technology, that made reprogramming controllable and generalizable to any cell type, and this new direction is just building upon, optimizing, and repurposing the technology for the ulterior motive of rejuvenation—comparably a less daunting task. We believe that as our understanding of aging become clearer, and hard evidence for age reversal becomes more prevalent, the dogma that aging is an immutable, irreversible process will be shattered. The field of medicine is fundamentally about challenging these limitations and revolutionizing the human condition. We believe the next revolution is upon us, as rejuvenation goes from mythology to gerontology.
\nResearch methodology is the path through which researchers need to conduct their research. It shows the path through which these researchers formulate their problem and objective and present their result from the data obtained during the study period. This research design and methodology chapter also shows how the research outcome at the end will be obtained in line with meeting the objective of the study. This chapter hence discusses the research methods that were used during the research process. It includes the research methodology of the study from the research strategy to the result dissemination. For emphasis, in this chapter, the author outlines the research strategy, research design, research methodology, the study area, data sources such as primary data sources and secondary data, population consideration and sample size determination such as questionnaires sample size determination and workplace site exposure measurement sample determination, data collection methods like primary data collection methods including workplace site observation data collection and data collection through desk review, data collection through questionnaires, data obtained from experts opinion, workplace site exposure measurement, data collection tools pretest, secondary data collection methods, methods of data analysis used such as quantitative data analysis and qualitative data analysis, data analysis software, the reliability and validity analysis of the quantitative data, reliability of data, reliability analysis, validity, data quality management, inclusion criteria, ethical consideration and dissemination of result and its utilization approaches. In order to satisfy the objectives of the study, a qualitative and quantitative research method is apprehended in general. The study used these mixed strategies because the data were obtained from all aspects of the data source during the study time. Therefore, the purpose of this methodology is to satisfy the research plan and target devised by the researcher.
The research design is intended to provide an appropriate framework for a study. A very significant decision in research design process is the choice to be made regarding research approach since it determines how relevant information for a study will be obtained; however, the research design process involves many interrelated decisions [1].
This study employed a mixed type of methods. The first part of the study consisted of a series of well-structured questionnaires (for management, employee’s representatives, and technician of industries) and semi-structured interviews with key stakeholders (government bodies, ministries, and industries) in participating organizations. The other design used is an interview of employees to know how they feel about safety and health of their workplace, and field observation at the selected industrial sites was undertaken.
Hence, this study employs a descriptive research design to agree on the effects of occupational safety and health management system on employee health, safety, and property damage for selected manufacturing industries. Saunders et al. [2] and Miller [3] say that descriptive research portrays an accurate profile of persons, events, or situations. This design offers to the researchers a profile of described relevant aspects of the phenomena of interest from an individual, organizational, and industry-oriented perspective. Therefore, this research design enabled the researchers to gather data from a wide range of respondents on the impact of safety and health on manufacturing industries in Ethiopia. And this helped in analyzing the response obtained on how it affects the manufacturing industries’ workplace safety and health. The research overall design and flow process are depicted in Figure 1.
Research methods and processes (author design).
To address the key research objectives, this research used both qualitative and quantitative methods and combination of primary and secondary sources. The qualitative data supports the quantitative data analysis and results. The result obtained is triangulated since the researcher utilized the qualitative and quantitative data types in the data analysis. The study area, data sources, and sampling techniques were discussed under this section.
According to Fraenkel and Warren [4] studies, population refers to the complete set of individuals (subjects or events) having common characteristics in which the researcher is interested. The population of the study was determined based on random sampling system. This data collection was conducted from March 07, 2015 to December 10, 2016, from selected manufacturing industries found in Addis Ababa city and around. The manufacturing companies were selected based on their employee number, established year, and the potential accidents prevailing and the manufacturing industry type even though all criterions were difficult to satisfy.
It was obtained from the original source of information. The primary data were more reliable and have more confidence level of decision-making with the trusted analysis having direct intact with occurrence of the events. The primary data sources are industries’ working environment (through observation, pictures, and photograph) and industry employees (management and bottom workers) (interview, questionnaires and discussions).
Desk review has been conducted to collect data from various secondary sources. This includes reports and project documents at each manufacturing sectors (more on medium and large level). Secondary data sources have been obtained from literatures regarding OSH, and the remaining data were from the companies’ manuals, reports, and some management documents which were included under the desk review. Reputable journals, books, different articles, periodicals, proceedings, magazines, newsletters, newspapers, websites, and other sources were considered on the manufacturing industrial sectors. The data also obtained from the existing working documents, manuals, procedures, reports, statistical data, policies, regulations, and standards were taken into account for the review.
In general, for this research study, the desk review has been completed to this end, and it had been polished and modified upon manuals and documents obtained from the selected companies.
The study population consisted of manufacturing industries’ employees in Addis Ababa city and around as there are more representative manufacturing industrial clusters found. To select representative manufacturing industrial sector population, the types of the industries expected were more potential to accidents based on random and purposive sampling considered. The population of data was from textile, leather, metal, chemicals, and food manufacturing industries. A total of 189 sample sizes of industries responded to the questionnaire survey from the priority areas of the government. Random sample sizes and disproportionate methods were used, and 80 from wood, metal, and iron works; 30 from food, beverage, and tobacco products; 50 from leather, textile, and garments; 20 from chemical and chemical products; and 9 from other remaining 9 clusters of manufacturing industries responded.
A simple random sampling and purposive sampling methods were used to select the representative manufacturing industries and respondents for the study. The simple random sampling ensures that each member of the population has an equal chance for the selection or the chance of getting a response which can be more than equal to the chance depending on the data analysis justification. Sample size determination procedure was used to get optimum and reasonable information. In this study, both probability (simple random sampling) and nonprobability (convenience, quota, purposive, and judgmental) sampling methods were used as the nature of the industries are varied. This is because of the characteristics of data sources which permitted the researchers to follow the multi-methods. This helps the analysis to triangulate the data obtained and increase the reliability of the research outcome and its decision. The companies’ establishment time and its engagement in operation, the number of employees and the proportion it has, the owner types (government and private), type of manufacturing industry/production, types of resource used at work, and the location it is found in the city and around were some of the criteria for the selections.
The determination of the sample size was adopted from Daniel [5] and Cochran [6] formula. The formula used was for unknown population size Eq. (1) and is given as
where n = sample size, Z = statistic for a level of confidence, P = expected prevalence or proportion (in proportion of one; if 50%, P = 0.5), and d = precision (in proportion of one; if 6%, d = 0.06). Z statistic (Z): for the level of confidence of 95%, which is conventional, Z value is 1.96. In this study, investigators present their results with 95% confidence intervals (CI).
The expected sample number was 267 at the marginal error of 6% for 95% confidence interval of manufacturing industries. However, the collected data indicated that only 189 populations were used for the analysis after rejecting some data having more missing values in the responses from the industries. Hence, the actual data collection resulted in 71% response rate. The 267 population were assumed to be satisfactory and representative for the data analysis.
The sample size for the experimental exposure measurements of physical work environment has been considered based on the physical data prepared for questionnaires and respondents. The response of positive were considered for exposure measurement factors to be considered for the physical environment health and disease causing such as noise intensity, light intensity, pressure/stress, vibration, temperature/coldness, or hotness and dust particles on 20 workplace sites. The selection method was using random sampling in line with purposive method. The measurement of the exposure factors was done in collaboration with Addis Ababa city Administration and Oromia Bureau of Labour and Social Affair (AACBOLSA). Some measuring instruments were obtained from the Addis Ababa city and Oromia Bureau of Labour and Social Affair.
Data collection methods were focused on the followings basic techniques. These included secondary and primary data collections focusing on both qualitative and quantitative data as defined in the previous section. The data collection mechanisms are devised and prepared with their proper procedures.
Primary data sources are qualitative and quantitative. The qualitative sources are field observation, interview, and informal discussions, while that of quantitative data sources are survey questionnaires and interview questions. The next sections elaborate how the data were obtained from the primary sources.
Observation is an important aspect of science. Observation is tightly connected to data collection, and there are different sources for this: documentation, archival records, interviews, direct observations, and participant observations. Observational research findings are considered strong in validity because the researcher is able to collect a depth of information about a particular behavior. In this dissertation, the researchers used observation method as one tool for collecting information and data before questionnaire design and after the start of research too. The researcher made more than 20 specific observations of manufacturing industries in the study areas. During the observations, it found a deeper understanding of the working environment and the different sections in the production system and OSH practices.
Interview is a loosely structured qualitative in-depth interview with people who are considered to be particularly knowledgeable about the topic of interest. The semi-structured interview is usually conducted in a face-to-face setting which permits the researcher to seek new insights, ask questions, and assess phenomena in different perspectives. It let the researcher to know the in-depth of the present working environment influential factors and consequences. It has provided opportunities for refining data collection efforts and examining specialized systems or processes. It was used when the researcher faces written records or published document limitation or wanted to triangulate the data obtained from other primary and secondary data sources.
This dissertation is also conducted with a qualitative approach and conducting interviews. The advantage of using interviews as a method is that it allows respondents to raise issues that the interviewer may not have expected. All interviews with employees, management, and technicians were conducted by the corresponding researcher, on a face-to-face basis at workplace. All interviews were recorded and transcribed.
The main tool for gaining primary information in practical research is questionnaires, due to the fact that the researcher can decide on the sample and the types of questions to be asked [2].
In this dissertation, each respondent is requested to reply to an identical list of questions mixed so that biasness was prevented. Initially the questionnaire design was coded and mixed up from specific topic based on uniform structures. Consequently, the questionnaire produced valuable data which was required to achieve the dissertation objectives.
The questionnaires developed were based on a five-item Likert scale. Responses were given to each statement using a five-point Likert-type scale, for which 1 = “strongly disagree” to 5 = “strongly agree.” The responses were summed up to produce a score for the measures.
The data was also obtained from the expert’s opinion related to the comparison of the knowledge, management, collaboration, and technology utilization including their sub-factors. The data obtained in this way was used for prioritization and decision-making of OSH, improving factor priority. The prioritization of the factors was using Saaty scales (1–9) and then converting to Fuzzy set values obtained from previous researches using triangular fuzzy set [7].
The researcher has measured the workplace environment for dust, vibration, heat, pressure, light, and noise to know how much is the level of each variable. The primary data sources planned and an actual coverage has been compared as shown in Table 1.
Planned versus actual coverage of the survey.
The response rate for the proposed data source was good, and the pilot test also proved the reliability of questionnaires. Interview/discussion resulted in 87% of responses among the respondents; the survey questionnaire response rate obtained was 71%, and the field observation response rate was 90% for the whole data analysis process. Hence, the data organization quality level has not been compromised.
This response rate is considered to be representative of studies of organizations. As the study agrees on the response rate to be 30%, it is considered acceptable [8]. Saunders et al. [2] argued that the questionnaire with a scale response of 20% response rate is acceptable. Low response rate should not discourage the researchers, because a great deal of published research work also achieves low response rate. Hence, the response rate of this study is acceptable and very good for the purpose of meeting the study objectives.
The pretest for questionnaires, interviews, and tools were conducted to validate that the tool content is valid or not in the sense of the respondents’ understanding. Hence, content validity (in which the questions are answered to the target without excluding important points), internal validity (in which the questions raised answer the outcomes of researchers’ target), and external validity (in which the result can generalize to all the population from the survey sample population) were reflected. It has been proved with this pilot test prior to the start of the basic data collections. Following feedback process, a few minor changes were made to the originally designed data collect tools. The pilot test made for the questionnaire test was on 10 sample sizes selected randomly from the target sectors and experts.
The secondary data refers to data that was collected by someone other than the user. This data source gives insights of the research area of the current state-of-the-art method. It also makes some sort of research gap that needs to be filled by the researcher. This secondary data sources could be internal and external data sources of information that may cover a wide range of areas.
Literature/desk review and industry documents and reports: To achieve the dissertation’s objectives, the researcher has conducted excessive document review and reports of the companies in both online and offline modes. From a methodological point of view, literature reviews can be comprehended as content analysis, where quantitative and qualitative aspects are mixed to assess structural (descriptive) as well as content criteria.
A literature search was conducted using the database sources like MEDLINE; Emerald; Taylor and Francis publications; EMBASE (medical literature); PsycINFO (psychological literature); Sociological Abstracts (sociological literature); accident prevention journals; US Statistics of Labor, European Safety and Health database; ABI Inform; Business Source Premier (business/management literature); EconLit (economic literature); Social Service Abstracts (social work and social service literature); and other related materials. The search strategy was focused on articles or reports that measure one or more of the dimensions within the research OSH model framework. This search strategy was based on a framework and measurement filter strategy developed by the Consensus-Based Standards for the Selection of Health Measurement Instruments (COSMIN) group. Based on screening, unrelated articles to the research model and objectives were excluded. Prior to screening, researcher (principal investigator) reviewed a sample of more than 2000 articles, websites, reports, and guidelines to determine whether they should be included for further review or reject. Discrepancies were thoroughly identified and resolved before the review of the main group of more than 300 articles commenced. After excluding the articles based on the title, keywords, and abstract, the remaining articles were reviewed in detail, and the information was extracted on the instrument that was used to assess the dimension of research interest. A complete list of items was then collated within each research targets or objectives and reviewed to identify any missing elements.
Data analysis method follows the procedures listed under the following sections. The data analysis part answered the basic questions raised in the problem statement. The detailed analysis of the developed and developing countries’ experiences on OSH regarding manufacturing industries was analyzed, discussed, compared and contrasted, and synthesized.
Quantitative data were obtained from primary and secondary data discussed above in this chapter. This data analysis was based on their data type using Excel, SPSS 20.0, Office Word format, and other tools. This data analysis focuses on numerical/quantitative data analysis.
Before analysis, data coding of responses and analysis were made. In order to analyze the data obtained easily, the data were coded to SPSS 20.0 software as the data obtained from questionnaires. This task involved identifying, classifying, and assigning a numeric or character symbol to data, which was done in only one way pre-coded [9, 10]. In this study, all of the responses were pre-coded. They were taken from the list of responses, a number of corresponding to a particular selection was given. This process was applied to every earlier question that needed this treatment. Upon completion, the data were then entered to a statistical analysis software package, SPSS version 20.0 on Windows 10 for the next steps.
Under the data analysis, exploration of data has been made with descriptive statistics and graphical analysis. The analysis included exploring the relationship between variables and comparing groups how they affect each other. This has been done using cross tabulation/chi square, correlation, and factor analysis and using nonparametric statistic.
Qualitative data analysis used for triangulation of the quantitative data analysis. The interview, observation, and report records were used to support the findings. The analysis has been incorporated with the quantitative discussion results in the data analysis parts.
The data were entered using SPSS 20.0 on Windows 10 and analyzed. The analysis supported with SPSS software much contributed to the finding. It had contributed to the data validation and correctness of the SPSS results. The software analyzed and compared the results of different variables used in the research questionnaires. Excel is also used to draw the pictures and calculate some analytical solutions.
The reliability of measurements specifies the amount to which it is without bias (error free) and hence ensures consistent measurement across time and across the various items in the instrument [8]. In reliability analysis, it has been checked for the stability and consistency of the data. In the case of reliability analysis, the researcher checked the accuracy and precision of the procedure of measurement. Reliability has numerous definitions and approaches, but in several environments, the concept comes to be consistent [8]. The measurement fulfills the requirements of reliability when it produces consistent results during data analysis procedure. The reliability is determined through Cranach’s alpha as shown in Table 2.
Internal consistency and reliability test of questionnaires items.
K stands for knowledge; M, management; T, technology; C, collaboration; P, policy, standards, and regulation; H, hazards and accident conditions; PPE, personal protective equipment.
Cronbach’s alpha is a measure of internal consistency, i.e., how closely related a set of items are as a group [11]. It is considered to be a measure of scale reliability. The reliability of internal consistency most of the time is measured based on the Cronbach’s alpha value. Reliability coefficient of 0.70 and above is considered “acceptable” in most research situations [12]. In this study, reliability analysis for internal consistency of Likert-scale measurement after deleting 13 items was found similar; the reliability coefficients were found for 76 items were 0.964 and for the individual groupings made shown in Table 2. It was also found internally consistent using the Cronbach’s alpha test. Table 2 shows the internal consistency of the seven major instruments in which their reliability falls in the acceptable range for this research.
Face validity used as defined by Babbie [13] is an indicator that makes it seem a reasonable measure of some variables, and it is the subjective judgment that the instrument measures what it intends to measure in terms of relevance [14]. Thus, the researcher ensured, in this study, when developing the instruments that uncertainties were eliminated by using appropriate words and concepts in order to enhance clarity and general suitability [14]. Furthermore, the researcher submitted the instruments to the research supervisor and the joint supervisor who are both occupational health experts, to ensure validity of the measuring instruments and determine whether the instruments could be considered valid on face value.
In this study, the researcher was guided by reviewed literature related to compliance with the occupational health and safety conditions and data collection methods before he could develop the measuring instruments. In addition, the pretest study that was conducted prior to the main study assisted the researcher to avoid uncertainties of the contents in the data collection measuring instruments. A thorough inspection of the measuring instruments by the statistician and the researcher’s supervisor and joint experts, to ensure that all concepts pertaining to the study were included, ensured that the instruments were enriched.
Insight has been given to the data collectors on how to approach companies, and many of the questionnaires were distributed through MSc students at Addis Ababa Institute of Technology (AAiT) and manufacturing industries’ experience experts. This made the data quality reliable as it has been continually discussed with them. Pretesting for questionnaire was done on 10 workers to assure the quality of the data and for improvement of data collection tools. Supervision during data collection was done to understand how the data collectors are handling the questionnaire, and each filled questionnaires was checked for its completeness, accuracy, clarity, and consistency on a daily basis either face-to-face or by phone/email. The data expected in poor quality were rejected out of the acting during the screening time. Among planned 267 questionnaires, 189 were responded back. Finally, it was analyzed by the principal investigator.
The data were collected from the company representative with the knowledge of OSH. Articles written in English and Amharic were included in this study. Database information obtained in relation to articles and those who have OSH area such as interventions method, method of accident identification, impact of occupational accidents, types of occupational injuries/disease, and impact of occupational accidents, and disease on productivity and costs of company and have used at least one form of feedback mechanism. No specific time period was chosen in order to access all available published papers. The questionnaire statements which are similar in the questionnaire have been rejected from the data analysis.
Ethical clearance was obtained from the School of Mechanical and Industrial Engineering, Institute of Technology, Addis Ababa University. Official letters were written from the School of Mechanical and Industrial Engineering to the respective manufacturing industries. The purpose of the study was explained to the study subjects. The study subjects were told that the information they provided was kept confidential and that their identities would not be revealed in association with the information they provided. Informed consent was secured from each participant. For bad working environment assessment findings, feedback will be given to all manufacturing industries involved in the study. There is a plan to give a copy of the result to the respective study manufacturing industries’ and ministries’ offices. The respondents’ privacy and their responses were not individually analyzed and included in the report.
The result of this study will be presented to the Addis Ababa University, AAiT, School of Mechanical and Industrial Engineering. It will also be communicated to the Ethiopian manufacturing industries, Ministry of Labor and Social Affair, Ministry of Industry, and Ministry of Health from where the data was collected. The result will also be availed by publication and online presentation in Google Scholars. To this end, about five articles were published and disseminated to the whole world.
The research methodology and design indicated overall process of the flow of the research for the given study. The data sources and data collection methods were used. The overall research strategies and framework are indicated in this research process from problem formulation to problem validation including all the parameters. It has laid some foundation and how research methodology is devised and framed for researchers. This means, it helps researchers to consider it as one of the samples and models for the research data collection and process from the beginning of the problem statement to the research finding. Especially, this research flow helps new researchers to the research environment and methodology in particular.
There is no “conflict of interest.”
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