\r\n\r\n \r\n\r\nVitamin K2 - Vital for Health and Wellbeing has been produced and distributed through the support from Kappa Bioscience, Norway.\r\n',isbn:"978-953-51-3020-8",printIsbn:"978-953-51-3019-2",pdfIsbn:"978-953-51-4895-1",doi:"10.5772/61430",price:139,priceEur:155,priceUsd:179,slug:"vitamin-k2-vital-for-health-and-wellbeing",numberOfPages:338,isOpenForSubmission:!1,isInWos:1,hash:"b2f9f024939ddc4f5da2a8afa3fcd9c9",bookSignature:"Jan Oxholm Gordeladze",publishedDate:"March 22nd 2017",coverURL:"https://cdn.intechopen.com/books/images_new/5169.jpg",numberOfDownloads:25033,numberOfWosCitations:7,numberOfCrossrefCitations:13,numberOfDimensionsCitations:25,hasAltmetrics:1,numberOfTotalCitations:45,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 6th 2015",dateEndSecondStepPublish:"January 31st 2016",dateEndThirdStepPublish:"March 25th 2016",dateEndFourthStepPublish:"May 30th 2016",dateEndFifthStepPublish:"August 31st 2016",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,editors:[{id:"36345",title:"Prof.",name:"Jan",middleName:"Oxholm",surname:"Gordeladze",slug:"jan-gordeladze",fullName:"Jan Gordeladze",profilePictureURL:"https://mts.intechopen.com/storage/users/36345/images/3823_n.jpg",biography:"Dr. Jan O. Gordeladze, Ph.D. (born 25th of April, 1950), holds a triple professor competence (Medical Biochemistry, Physiology, and Pharmacology), and is presently working as a Professor Emeritus at the Department of Biochemistry, Institute of Basic Medical Science, University of Oslo, Norway. He has previously been employed as the Medical Director of MSD, Norway, serving two years as a Fulbright scholar at the NIH, Bethesda, Maryland, USA. From 2006-2009 he was employed as Associate Professor at the University of Montpellier, France. He is a member of the Norwegian Stem Cell Center, and his research has over the past 7-10 years been devoted to differentiation of osteochondral cells from stem cells focusing on the impact of transcription factors and microRNA species constituting regulatory loop interactions with functional target genes. He has published more than 120 scientific articles, reviews/book chapters and presented more than 250 abstracts/posters/talks at conferences worldwide. 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He has been Chief of Gastroenterology Service at HUCA Hospital in Oviedo for more than 40 years. He obtained a PhD in 1975 and has had a long teaching and research career. He has published 590 scientific papers in both English and Spanish and written thirty-five book chapters and edited twenty-four books. Dr. Rodrigo has participated as a main investigator in forty-five clinical trials. He has directed forty doctoral theses and contributed to the formation of 100 specialists in gastroenterology working in hospitals in Spain and abroad. 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1. Introduction
Olive allergen concentrations in the pollen grain are critical in allergic response of atopic patients. Existing evidence shows that climatic factors can influence pollen allergen content. To date, the influences of temperature and precipitations over the content of birch and olive pollens in their respective major allergens have been analyzed. Bet v 1, the major allergen of birch pollen, increased its expression (Buters et al. 2008) and presented a higher allergenicity (Ahlholm et al. 1998) while temperatures were elevated. Differently, the content in the major olive pollen allergen Ole e 1 showed no apparent correlation with either temperatures or precipitations (Fernández Caldas et al. 2007). However, a positive correlation between total allergenicity and rainfall occurring in winter months was found. This correlation was not analyzed independently for each pollen allergen.
Our aim was to extend the observations carried out in olive pollen, by analyzing the effects of climate parameters over the expression of four olive pollen allergens. Using a recently developed multiplex western blotting system for the assessment of allergenic molecules (Morales et al. 2012), we have simultaneously detected and quantified two major (Ole e 1 and Ole e 9) and two minor allergens (Ole e 2 and Ole e 5) in the pollen extracts from seven olive cultivars collected along 4-7 consecutive years. The considered climatic variables included temperature, precipitation, number of rain days and humidity. Data were provided by the Spanish network for the temporal observation of ecosystems (REDOTE). Correlations between the allergen contents (both individually and all inclusive) and climate variables were studied by applying Spearman correlation tests. Results showed significant variations in the expression of the four allergens in the seven cultivars throughout the years of the analysis. All these positive correlations corresponded exactly to the period of time starting the winter prior to each flowering period to the end of period (this is, December from the previous year to June).
These results are discussed as regard to their putative incidence in the development of symptoms by patients, the requirements for medical assistance and the rates of admission into clinical centres and hospitals. Agronomical implications in olive sexual reproduction, including fruit setting and fruit production, are also discussed.
2. Materials and methods
2.1. Pollen samples
Olea europaea L. pollen samples were obtained during May and June of 2000-2007 from cultivated trees of the cultivars: ‘Arbequina’, ‘Blanqueta’, ‘Hojiblanca’, ‘Manzanilla de Sevilla’, ‘Picual’, ‘Verdial de Huévar’ and ‘Verdial de Vélez’ situated at the IFAPA center “Alameda del Obispo” (Andalusian Regional Government, Córdoba, Spain). Pollen samples were collected from numerous branches of at least two trees of each cultivar by shaking flowering shoots inside paper bags. Prior to its storage in liquid nitrogen, the harvested pollen was sieved through a 150 µm mesh in order to eliminate fallen corollas, anthers and other rests. After light microscopy observation, foreign-species pollen was estimated to be <0.1% and other plant parts <0.5% for all the cultivars used.
2.2. Preparation of crude protein extracts and SDS-PAGE
Crude protein extracts were obtained by stirring 1 g of pollen for each cultivar in 10 ml extraction buffer (0.01 M ammonium bicarbonate, pH 8.0, and 2 mM phenylmethylsulfonyl fluoride) for 8 h at 4°C. After centrifugation (2 x 30 minutes at 14,000 rpm at 4°C), the supernatants were filtered through a 0.2 µm filter, and stored in aliquots at –20°C. Protein concentration in the different samples was measured using the Bio-Rad reagent (Bio-Rad, Hercules, CA, USA) and bovine serum albumin (BSA) as standard.
Proteins (30 µg per lane) and Mw1 (New England BioLabs, Ipswich, MA, USA) and Mw2 standards (MBI Fermentas, Vilnius, Lithuania) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in 15% gels in a MiniProtean II system (Bio-Rad). The resulting gels were stained with silver nitrate (Rabilloud et al., 1994).
2.3. Immunoblotting
Gels obtained as described above were transferred onto BioTrace® polyvinylidene difluoride (PVDF) membranes (Pall BioSupport, Port Washington, NY, USA) at 100 V for 1.5 hours using a Mini Trans-Blot Electrophoretic Transfer Cell (Bio-Rad). Prior to the treatment with antibodies, the membranes were blocked with TBST buffer (Tris buffered saline: TBS + 0.3% v/v Tween 20) + 10% w/v dried skimmed milk. For membrane probing, a recently developed multiplex Western blotting system for the assessment of allergenic molecules (Morales et al. 2012) has been used. Briefly, the simultaneous detection of four olive pollen allergens (Ole e 1, Ole e 2, Ole e 5 and Ole e 9) on a single blot using a monoclonal antibody from mouse and three polyclonal antibodies raised in rabbit is carried out. We utilized unconjugated Fab antibody fragments for blocking rabbit primary antibodies, and fluorescence-based detection. These changes allowed an accurate and reliable comparative quantitation of these allergens among pollen protein samples.
2.4. Absolute and relative quantitation of allergens
Imaging was carried out with a Pharos FX Plus Molecular Imager (Bio-Rad) using the Quantity One v4.6.2 software (Bio-Rad). The intensity of each fluorescent band was calculated using the quantitation tools of the Quantity One v4.6.2 software.
2.5. Climate parameters
Climate parameters were obtained from the weather tracking station situated at Córdoba airport (Spain) during the years 2000 to 2007. Information was extracted from the Spanish long term ecological research network (REDOTE).
2.6. Statistical analysis
Continuous variables were subjected to a study of normality distribution by using the Kolmorov-Smirnov test. Associations between continuous variables (non-normal distribution) were described by analysis of bivariate Spearman correlation (two-tailed). Significant correlation was considered P<0.05.
3. Results
3.1. Multiplex determination of year-to-year allergen expression in seven olive cultivars
Figure 1A shows the protein profiles of the extracts analyzed after SDS-PAGE and silver staining. The different panels correspond to the cultivars analyzed between the years 2000 and 2007. Bands were observed in the range of 15 to 75 kDa. Clear quantitative differences were distinguished, from which the most conspicuous were those in the bands of 20 kDa and 18.4 kDa among de different years, with the exception of cultivars ‘Arbequina’, ‘Verdial de Huévar’ and ‘Hojiblanca’, in which the Ole e 1 bands were not well identified after silver staining. The remaining allergens studied were not identified after silver staining.
Allergenic profiles were assessed by multiplex immunotransference (Figure 1B). The cultivars ‘Arbequina’ and ‘Hojiblanca’ displayed two Ole e 1 forms (20 kDa and 18.4 kDa) in contrast with the remaining cultivars, were the three major forms of Ole e 1 (22 kDa, 20 kDa and 18.4 kDa) were distinguished. Differences in the expression of this allergen depending on the year of pollen collection can be easily noticed.
Ole e 2 was visualized in all cultivars in the form of two bands with sizes of 15.1 kDa and 14 kDa, with the exception of the ‘Verdial de Vélez’ cultivar, in which only the 15.1 kDa band was present. As described for Ole e 1, the presence of noticeable differences in the expression of the Ole e 2 allergen, depending the year of pollen collection can be observed in the corresponding multiplex immunoblots.
Figure 1.
A) Silver staining of total protein extracts obtained from pollen corresponding to the seven cultivars analyzed trhoughout the different years. B) Multiplex detection of the allergens Ole e 1, Ole e 2, Ole e 5 and Ole e 9 in the same extracts described in panel A. Thirty micrograms of total protein were loaded in each lane. Lane numbers (0-7) correspond to the years analyzed (2000-2007). Molecular weight standards are displayed on the left.
Ole e 5 allergen can be observed in the cultivars analyzed. However, only a weak signal was detected in the cultivars ‘Hojiblanca’ and ‘Verdial de Huévar’. Year-to-year differences in the expression of this allergen are clearly observed in ‘Picual’ and ‘Blanqueta’ cultivars.
Finally, Ole e 9 was expressed in all cultivars analyzed. Year-to-year differences in their expression are observed as well, with the sharpest differences in the cultivars ‘Blanqueta’ and ‘Manzanilla de Sevilla’.
3.2. Involvement of climatic factors in the expression of Ole e 1 allergen
Correlation between the data corresponding to the major climatic factors and the expression of the allergens Ole e 1, Ole e 2, Ole e 5 y Ole e 9 was individually assessed for each cultivar by means of Spearman correlation analyses. The analysis of the correlation between the levels of the Ole e 1 and these climatic parameters is shown in Table 1. The presence of a statistically significant positive correlation between the expression of the different Ole e 1 forms (either individually or their added values) and the accumulated yearly average temperatures (the addition of the monthly average temperatures during those months prior to the blooming period -July of the prior year to June of the current year-) was determined for the cultivar ‘Blanqueta’. Total levels of the Ole e 1 allergen correlated positively with the monthly average temperatures during the winter period (for the cultivar ‘Verdial de Huévar’) or the spring period (cultivars ‘Blanqueta’, ‘Hojiblanca’).
Analysis of bivariate Spearman correlation (two-tailed) between the expression of the different Ole e 1 forms both individually and jointly (CNT*mm2) and different climatic factors. R: Spearman correlation coefficient. * Statistically significant correlation P<0.05; ** P<0.01.
Moreover, statistically significant positive correlations were observed between Ole e 1 expression and minimum temperatures (cultivars ‘Blanqueta’, ‘Hojiblanca’, ‘Picual’ and ‘Manzanilla de Sevilla’) or maximum temperatures (cultivars ‘Picual’ and ‘Manzanilla de Sevilla’) during the spring period.
Total expression of Ole e 1 correlated positively with the added monthly precipitation occurred during the winter and spring period (cultivars ‘Hojiblanca’ and ‘Verdial de Huévar’), as well as with the total precipitation occurred during the months prior to flowering (cultivar ‘Manzanilla de Sevilla’). Positive correlations are also observed between total levels of Ole e 1 allergen and precipitation, number of days of rain, and the monthly average of relative humidity during the months prior to the blooming season.
3.3. Involvement of climatic factors in the expression of Ole e 2 allergen
Table 2 displays the observed correlations between Ole e 2 and different climatic factors.
Analysis of bivariate Spearman correlation (two-tailed) between the expression of the different Ole e 2 forms both individually and jointly (CNT*mm2) and different climatic factors. R: Spearman correlation coefficient. * Statistically significant correlation P<0.05; ** P<0.01. AT: average temperature. mT: minimum temperature. MT: maximum temperature. mRH: monthly relative humidity.
The expression levels of the bands corresponding to the Ole e 2 allergen (Table 2) and/or their addition, correlated positively with the average temperatures for particular months during the winter period (cultivars ‘Hojiblanca’ and ‘Verdial de Huévar’) or spring (cultivar ‘Arbequina’). Statistically significant correlations were also observed between the expression of Ole e 2 and the maximum/minimum temperatures over the months corresponding to the spring (cultivars ‘Arbequina’ and ‘Hojiblanca’).
The expression of one of the Ole e 2 form is positively correlated to the accumulated monthly precipitation occurred during the winter months for the cultivar ‘Arbequina’, as well as to the precipitation occurred in particular months during the winter and spring (cultivars ‘Arbequina’, ‘Verdial de Huévar’ and ‘Manzanilla de Sevilla’). The Ole e 2 form with the higher molecular weight (15.1 kDa) also presented correlation to the accumulated monthly relative humidity during the months of winter and spring (cultivars ‘Hojiblanca’ and ‘Verdial de Huévar’). Similarly, positive correlation was detected between the total levels of Ole e 2 and the total number of days of rain over the winter and spring months (cultivar ‘Verdial de Huévar’).
3.4. Involvement of the climatic factors in the expression of Ole e 5 allergen
As respects to Ole e 5 allergen (Table 3), the presence of a statistically significant positive correlation was determined between the minimum temperature during May, and the precipitation observed during February in the ‘Blanqueta’ cultivar. In the cultivar ‘Verdial de Vélez’, Ole e 5 correlated to the number of days of rain and the relative humidity over the months of February and April.
Cultivar
Climatic factor
Ole e 5
Blanqueta
mT May (°C)
R
.9*
P
.0187
precipitation February (mm)
R
.9 *
P
.0037
Verdial de Vélez
No. days of rain in February
R
1**
P
<0.001
No. days of rain in April
R
1**
P
<0.001
mRH February (%)
R
1**
P
<0.001
mRH April (%)
R
1**
P
<0.001
Table 3.
Analysis of bivariate Spearman correlation (two-tailed) between the expression of Ole e 5 (CNT*mm2) and different climatic factors. R: Spearman correlation coefficient. *Statistically significant correlation P<0.05; ** P<0.01. AT: average temperature. mT: minimum temperature. MT: maximum temperature. mRH: monthly relative humidity.
3.5. Involvement of climatic factors in the expression of Ole e 9 allergen
As regard to the Ole e 9 allergen, its expression correlated positively (Table 4) with the average temperatures during the months of January (cultivars ‘Arbequina’, and ‘Verdial de Huévar’) and March (‘Hojiblanca’). In the same way, a correlation was demonstrated between Ole e 9 expression and the number of days of rain during March in the cultivars ‘Arbequina’ and ‘Verdial de Vélez’. The average relative humidity also showed a statistically relevant correlation during the months of winter, for the cultivar ‘Picual’, and specifically for January in the cultivar ‘Verdial de Huévar’. In addition, and for this last cultivar ‘Verdial de Huévar’, the relative humidity presents a positive correlation during the spring months of April and May.
3.6. Several examples of data distributions
We describe next several examples of data distributions corresponding to climatic factors showing a statistically significant correlation with the expression level of the different allergens. Charts represent the allergen levels (represented as band intensities) plotted against climatic factors like temperature (Figure 2), precipitation, relative humidity and number of days of rain (Figure 3). A reference line showing a theoretical absolute linear correlation between the variables was added to the figures.
Cultivar
Climatic factor
Ole e 9
Arbequina
AT January (°C)
R
.829*
P
.042
No of days of rain in March
R
.088*
P
.02
Hojiblanca
AT March (°C)
R
1**
P
<0.001
mT March (°C)
R
1**
P
<0.001
Picual
mRH December (%)
R
.857*
P
.014
Verdial de Vélez
No. days of rain in March
R
.949
P
.05
Verdial de Huévar
AT January (°C)
R
1**
P
<0.001
No. days of rain in February
R
.949
P
.05
mRH January (%)
R
1**
P
<0.001
mRH April (%)
R
1**
P
<0.001
mRH May (%)
R
1**
P
<0.001
Table 4.
Analysis of bivariate Spearman correlation (two-tailed) between the expression of Ole e 9 (CNT*mm2) and different climatic factors. R: Spearman correlation coefficient. *Statistically significant correlation P<0.05; ** P<0.01. AT: average temperature. mT: minimum temperature. MT: maximum temperature. mRH: monthly relative humidity.
Figure 2.
Examples of charts displaying data distribution referring to temperatures plotted against the expression levels of the following allergens: A) Ole e 1, B) Ole e 2, C) Ole e 5 and D) Ole e 9.
Figure 3.
Examples of charts displaying data distribution referring to precipitation, relative humidity and number of days of rain plotted against the expression levels of the following allergens: A) Ole e 1, B) Ole e 2, C) Ole e 5 and D) Ole e 9.
4. Discussion and conclusions
The allergen content of the olive pollen is clearly influenced by the genetic origin of the cultivar analyzed (Hamman Khalifa et al. 2008). Diverse authors have also indicated that climatic factors may have different effects over olive flowering and pollen development (Galan et al. 2001; Orlandi et al. 2010), as well as over olive pollen allergenicity (Fernández-Caldas et al. 2007).
The results presented here demonstrate that changes occur in the expression of four relevant allergens in seven cultivars, and throughout consecutive years. However, results are considered heterogeneous between the different cultivars. The climatic parameters taken into account were temperature, precipitation, number of days of rain or relative humidity during the months prior to the flowering period, which normally corresponds to the months of May and June.
As regard to the temperature, correlations were not observed for a particular month or for the accumulated yearly temperature in all cultivars in a simultaneous way. However, the point correlations detected always corresponded to the winter and summer months (January-June) prior to the flowering period. This can be due to the intrinsic characteristics and requirements of each cultivar, or to the low number of cultivars and years studied, which may limit the statistical power of the analysis.
This results are in good agreement with those described for the birch major pollen allergen Bet v 1 (Buters et al. 2008), where an increase of this allergen was observed in higher temperatures, determined by growth of sampling trees in different climatic regions, over two consecutive years. Another study suggests a higher allergenic response to the Bet v 1 allergen present in birch pollen extracts grown in high average daily temperatures, in comparison to trees cultivated at lower temperatures (Ahlholm et al. 1998). This assay was carried out by using sera from patients sensitive to this pollen by using immunoblotting experiments. Our results indicate that high temperatures also seem to exert an increase in the expression of allergens in certain olive cultivars.
We may extrapolate that an increase in the average temperatures would bring a parallel increase in pollen allergenic content and allergenicity as a consequence. This could be therefore one of the consequences of the so-called global warming produced by the greenhouse effect (Ahlholm et al. 1998; Cecchi et al. 2010). Moreover, we may speculate that the fact that urban areas are often exposed to temperatures 2 to 8 °C higher that rural areas (Oke 1987) may result in increased olive pollen allergenicity and allergy prevalence in and around the cities, where the olive tree is also present, as it is widely used as an ornamental plant.
Fernández-Caldas et al. (2007) performed a similar study using six olive cultivars, over a period of five consecutive years. This study analyzed the content in the olive pollen major allergen (Ole e 1) only, which did not show statistically significant correlations to the precipitation average, nor to the average temperatures over the winter months prior to flowering. These authors described, however, a positive correlation between overall allergenicity and the average precipitation occurred during the winter months, for all the cultivars analyzed.
Several relevant differences are noticeable between the present study and that of Fernández-Caldas et al. (2007). First, this work combines the information available on four allergens, Ole e 1, Ole e 2, Ole e 5 and Ole e 9, instead of the olive pollen major allergen only. Therefore, the conclusions obtained are more similar to those described by Fernández-Caldas et al. referring to whole olive pollen allergenicity than those referring to Ole e 1 only. On the other hand, the present study uses accumulated data instead of averages for parameters like precipitation and number of monthly/seasonal/yearly days of rain. Moreover, although in some cases, monthly averages of temperature and relative humidity were considered, specific correlations were calculated upon the accumulated data for seasonal/yearly periods.
From an allergenic point of view, the present results suggest that those years with high levels of precipitation and high temperatures during the winter and the spring periods, contribute an enhanced allergenic content of the pollen grains at the end of the spring. We also may postulate that those areas of olive culture which are supplemented by artificial watering may yield a higher allergenicity for olive pollen-sensitized patients. The same would apply to urban areas, where ornamental olive trees are usually watered.
From an agronomic point of view, the described increase in the expression of the allergen Ole e 1 which takes place under increased temperature and precipitation might result in important differences in the reproductive behavior of the plant. Thus, preliminary observations (Morales et al. unpublished), indicated that the enhanced levels of Ole e 1 correlated to increased rates of pollen germination and lower degree of olive self-incompatibility. This hypothesis is also in good agreement with experimental observations made by several authors as regard to the effects of climatic factor on self-pollination and productivity in the olive (Androulakis and Loupassaki 1990, Lavee et al. 2002).
Acknowledgement
This work was supported by the Spanish Ministry of Science and Innovation (MICINN) (ERDF-cofinanced projects AGL2008-00517, BFU2011-22779 and PIE-200840I186) and the Junta de Andalucía (ERDF-cofinanced projects P2010-CVI5767 and P2010-AGR6274). The authors acknowledge the availability of plant material and the collaboration of the staff of the IFAPA center “Venta del Llano” (Mengíbar, Spain) depending from the Andalusian Regional Government.
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Introduction",level:"1"},{id:"sec_2",title:"2. Materials and methods ",level:"1"},{id:"sec_2_2",title:"2.1. Pollen samples",level:"2"},{id:"sec_3_2",title:"2.2. Preparation of crude protein extracts and SDS-PAGE ",level:"2"},{id:"sec_4_2",title:"2.3. Immunoblotting ",level:"2"},{id:"sec_5_2",title:"2.4. Absolute and relative quantitation of allergens ",level:"2"},{id:"sec_6_2",title:"2.5. Climate parameters ",level:"2"},{id:"sec_7_2",title:"2.6. Statistical analysis ",level:"2"},{id:"sec_9",title:"3. Results ",level:"1"},{id:"sec_9_2",title:"3.1. Multiplex determination of year-to-year allergen expression in seven olive cultivars",level:"2"},{id:"sec_10_2",title:"3.2. Involvement of climatic factors in the expression of Ole e 1 allergen ",level:"2"},{id:"sec_11_2",title:"3.3. Involvement of climatic factors in the expression of Ole e 2 allergen",level:"2"},{id:"sec_12_2",title:"3.4. Involvement of the climatic factors in the expression of Ole e 5 allergen",level:"2"},{id:"sec_13_2",title:"3.5. Involvement of climatic factors in the expression of Ole e 9 allergen",level:"2"},{id:"sec_14_2",title:"3.6. Several examples of data distributions",level:"2"},{id:"sec_16",title:"4. Discussion and conclusions",level:"1"}],chapterReferences:[{id:"B1",body:'AhlholmJ. U.HelanderM. L.SavolainenJ.\n\t\t\t\t\t1998 Genetic and environmental factors affecting the allergenicity of birch (Betula pubescens ssp. czerepanovii [Orl.] Hamet-ahti) pollen. Clinical and Experimental Allergy, 28\n\t\t\t\t\t11\n\t\t\t\t\t13841388 , 1365-2222'},{id:"B2",body:'AndroulakisI. I.LoupassakiM. H.\n\t\t\t\t\t1990 Studies on the self-fertility of some cultivars in the area of Crete. Acta Horticulturae, 286\n\t\t\t\t\t159162 , 0567-7572'},{id:"B3",body:'ButersJ. T.KascheA.WeichenmeieraI.SchoberW.KlausS.Traidl-HoffmannC.MenzelA.Huss-MarpJ.KrämerU.BehrendtH.\n\t\t\t\t\t2008 Year-to-year variation in release of Bet v 1 allergen from birch pollen: evidence for geographical differences between West and South Germany. International Archives of Allergy & Immunology, 145\n\t\t\t\t\t2\n\t\t\t\t\t122130 , 1018-2438'},{id:"B4",body:'CecchiL.D’AmatoG.AyresJ. G.GalanC.ForastiereF.ForsbergB.GerritsenJ.NunesC.BehrendtH.AkdisC.DahlR.Annesi-MaesanoI.\n\t\t\t\t\t2010 Projections of the effects of climate change on allergic asthma: the contribution of aerobiology. Allergy, 65\n\t\t\t\t\t9\n\t\t\t\t\t10731081 , 0105-4538'},{id:"B5",body:'Fernández-CaldasE.CarnésJ.IraolaV.CasanovasM.\n\t\t\t\t\t2007 Comparison of the allergenicity and Ole e 1 content of 6 varieties of Olea euroapea pollen collected during 5 consecutive years. Annals of Allergy, Asthma & Immunology, 98\n\t\t\t\t\t5\n\t\t\t\t\t464470 , 1081-1206'},{id:"B6",body:'GalanC.CariñanosP.Garcia-MazoH.AlcazarP.Dominguez-VilchesE.\n\t\t\t\t\t2001 Model for forecasting Olea europaea L. airborne pollen in South-West Andalusia, Spain. International Journal of Biometeorology, 45\n\t\t\t\t\t2\n\t\t\t\t\t5963 , 0020-7128'},{id:"B7",body:'HammanKhalifa. A. M.CastroA. J.RodríguezGarcía. M. I.AlchéJ. D.\n\t\t\t\t\t2008 Olive cultivar origin is a major cause of polymorphism for Ole e 1 pollen allergen. BMC Plant Biology, 8\n\t\t\t\t\t10\n\t\t\t\t\t1471-2229'},{id:"B8",body:'LaveeS.TaryanJ.LevinJ.HaskalA.\n\t\t\t\t\t2002 Importancia de la polinización cruzada en distintas variedades de olivo cultivadas en olivares intensivos de regadío. Olivae: revista oficial del Consejo Oleícola Internacional, 91\n\t\t\t\t\t2536 , 0025-5996X'},{id:"B9",body:'OkeT. R.\n\t\t\t\t\t1987 Boundary Layer Climates. 2nd. Edition. London: Routledge. 435 pp.'},{id:"B10",body:'OrlandiF.SgromoC.BonofiglioT.RugaL.RomanoB.FornaciariM.\n\t\t\t\t\t2010 Spring Influences on Olive Flowering and Threshold Temperatures Related to Reproductive Structure Formation. HortScience, 45\n\t\t\t\t\t7\n\t\t\t\t\t10521057 , 0018-5345'},{id:"B11",body:'MoralesS.CastroA. J.Jiménez-LópezJ. C.FloridoF.Rodríguez-GarcíaM. I.AlchéJ. D.\n\t\t\t\t\t2012 A novel multiplex method for the simultaneous detection and relative quantification of pollen allergens. Electrophoresis, 33\n\t\t\t\t\t18 , DOI 10.1002/elps.201100667, 1522-2683'},{id:"B12",body:'RabilloudT.VuillardL.GillyC.LawrenceJ. J.\n\t\t\t\t\t1994 Silver-staining of proteins in polyacrylamide gels: a general overview. Cellular and Molecular Biology (Noisy-le-Grand, France), 40\n\t\t\t\t\t1\n\t\t\t\t\t5775 , 0116-5158X'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Sonia Morales",address:null,affiliation:'
Proteomic Research Service, Hospital Universitario San Cecilio, Granada,, Spain
Estación Experimental del Zaidín (CSIC), Granada,, Spain
Estación Experimental del Zaidín (CSIC), Granada,, Spain
'},{corresp:null,contributorFullName:"Juan de Dios Alché",address:null,affiliation:'
Estación Experimental del Zaidín (CSIC), Granada,, Spain
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1. Introduction
Research 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.
2. Research design
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.
Figure 1.
Research methods and processes (author design).
3. Research methodology
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.
3.1 The study area
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.
3.2 Data sources
3.2.1 Primary data sources
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).
3.2.2 Secondary data
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.
4. Population and sample size
4.1 Population
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.
4.2 Questionnaire sample size determination
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
E1
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.
4.3 Workplace site exposure measurement sample determination
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.
5. Data collection methods
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.
5.1 Primary data collection methods
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.
5.1.1 Workplace site observation data collection
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.
5.1.2 Data collection through interview
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.
5.1.3 Data collection through questionnaires
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.
5.1.4 Data obtained from experts’ opinion
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].
5.1.5 Workplace site exposure measurement
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.
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.
5.1.6 Data collection tool pretest
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.
5.2 Secondary data collection methods
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.
6. Methods of data analysis
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.
6.1 Quantitative data analysis
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.
6.2 Qualitative data analysis
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.
6.3 Data analysis software
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.
7. The reliability and validity analysis of the quantitative data
7.1 Reliability of data
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.
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.
7.2 Reliability analysis
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.
7.3 Validity
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.
8. Data quality management
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.
9. Inclusion criteria
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.
10. Ethical consideration
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.
11. Dissemination and utilization of the result
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.
12. Conclusion
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.
Conflict of interest
There is no “conflict of interest.”
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New York: John Wiley & Sons Inc; 2000'},{id:"B2",body:'Saunders M, Lewis P, Thornhill A. Research Methods for Business Student. 5th ed. Edinburgh Gate: Pearson Education Limited; 2009'},{id:"B3",body:'Miller P. Motivation in the Workplace. Work and Organizational Psychology. Oxford: Blackwell Publishers; 1991'},{id:"B4",body:'Fraenkel FJ, Warren NE. How to Design and Evaluate Research in Education. 4th ed. New York: McGraw-Hill; 2002'},{id:"B5",body:'Danniel WW. Biostatist: A Foundation for Analysis in the Health Science. 7th ed. New York: John Wiley & Sons; 1999'},{id:"B6",body:'Cochran WG. Sampling Techniques. 3rd ed. New York: John Wiley & Sons; 1977'},{id:"B7",body:'Saaty TL. The Analytical Hierarchy Process. Pittsburg: PWS Publications; 1990'},{id:"B8",body:'Sekaran U, Bougie R. Research Methods for Business: A Skill Building Approach. 5th ed. New Delhi: John Wiley & Sons, Ltd; 2010. pp. 1-468'},{id:"B9",body:'Luck DJ, Rubin RS. Marketing Research. 7th ed. New Jersey: Prentice-Hall International; 1987'},{id:"B10",body:'Wong TC. Marketing Research. Oxford, UK: Butterworth-Heinemann; 1999'},{id:"B11",body:'Cronbach LJ. Coefficient alpha and the internal structure of tests. Psychometrika. 1951;16:297-334'},{id:"B12",body:'Tavakol M, Dennick R. Making sense of Cronbach’s alpha. International Journal of Medical Education. 2011;2:53-55. DOI: 10.5116/ijme.4dfb.8dfd'},{id:"B13",body:'Babbie E. The Practice of Social Research. 12th ed. Belmont, CA: Wadsworth; 2010'},{id:"B14",body:'Polit DF, Beck CT. Generating and Assessing Evidence for Nursing Practice. 8th ed. Williams and Wilkins: Lippincott; 2008'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Kassu Jilcha Sileyew",address:"jkassu@gmail.com",affiliation:'
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El-Hemaly\nProfessor OB/GYN & Urogynecology\nFaculty of medicine, Al-Azhar University \nPersonal Information: \nMarried with two children\nWife: Professor Laila A. Moussa MD.\nSons: Mohamad A. M. El-Hemaly Jr. MD. Died March 25-2007\nMostafa A. M. El-Hemaly, Computer Scientist working at Microsoft Seatle, USA. \nQualifications: \n1.\tM.B.-Bch Cairo Univ. June 1963. \n2.\tDiploma Ob./Gyn. Cairo Univ. April 1966. \n3.\tDiploma Surgery Cairo Univ. Oct. 1966. \n4.\tMRCOG London Feb. 1975. \n5.\tF.R.C.S. Glasgow June 1976. \n6.\tPopulation Study Johns Hopkins 1981. \n7.\tGyn. Oncology Johns Hopkins 1983. \n8.\tAdvanced Laparoscopic Surgery, with Prof. Paulson, Alexandria, Virginia USA 1993. \nSocieties & Associations: \n1.\t Member of the Royal College of Ob./Gyn. London. \n2.\tFellow of the Royal College of Surgeons Glasgow UK. \n3.\tMember of the advisory board on urogyn. FIGO. \n4.\tMember of the New York Academy of Sciences. \n5.\tMember of the American Association for the Advancement of Science. \n6.\tFeatured in �Who is Who in the World� from the 16th edition to the 20th edition. \n7.\tFeatured in �Who is Who in Science and Engineering� in the 7th edition. \n8.\tMember of the Egyptian Fertility & Sterility Society. \n9.\tMember of the Egyptian Society of Ob./Gyn. \n10.\tMember of the Egyptian Society of Urogyn. \n\nScientific Publications & Communications:\n1- Abdel Karim M. El Hemaly*, Ibrahim M. Kandil, Asim Kurjak, Ahmad G. Serour, Laila A. S. Mousa, Amr M. Zaied, Khalid Z. El Sheikha. \nImaging the Internal Urethral Sphincter and the Vagina in Normal Women and Women Suffering from Stress Urinary Incontinence and Vaginal Prolapse. Gynaecologia Et Perinatologia, Vol18, No 4; 169-286 October-December 2009.\n2- Abdel Karim M. El Hemaly*, Laila A. S. Mousa Ibrahim M. Kandil, Fatma S. El Sokkary, Ahmad G. Serour, Hossam Hussein.\nFecal Incontinence, A Novel Concept: The Role of the internal Anal sphincter (IAS) in defecation and fecal incontinence. Gynaecologia Et Perinatologia, Vol19, No 2; 79-85 April -June 2010.\n3- Abdel Karim M. El Hemaly*, Laila A. S. Mousa Ibrahim M. Kandil, Fatma S. El Sokkary, Ahmad G. Serour, Hossam Hussein.\nSurgical Treatment of Stress Urinary Incontinence, Fecal Incontinence and Vaginal Prolapse By A Novel Operation \n"Urethro-Ano-Vaginoplasty"\n Gynaecologia Et Perinatologia, Vol19, No 3; 129-188 July-September 2010.\n4- Abdel Karim M. El Hemaly*, Ibrahim M. Kandil, Laila A. S. Mousa and Mohamad A.K.M.El Hemaly.\nUrethro-vaginoplasty, an innovated operation for the treatment of: Stress Urinary Incontinence (SUI), Detursor Overactivity (DO), Mixed Urinary Incontinence and Anterior Vaginal Wall Descent. \nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/ urethro-vaginoplasty_01\n\n5- Abdel Karim M. El Hemaly, Ibrahim M Kandil, Mohamed M. Radwan.\n Urethro-raphy a new technique for surgical management of Stress Urinary Incontinence.\nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/\nnew-tech-urethro\n\n6- Abdel Karim M. El Hemaly, Ibrahim M Kandil, Mohamad A. Rizk, Nabil Abdel Maksoud H., Mohamad M. Radwan, Khalid Z. El Shieka, Mohamad A. K. M. El Hemaly, and Ahmad T. El Saban.\nUrethro-raphy The New Operation for the treatment of stress urinary incontinence, SUI, detrusor instability, DI, and mixed-type of urinary incontinence; short and long term results. \nhttp://www.obgyn.net/urogyn/urogyn.asp?page=urogyn/articles/\nurethroraphy-09280\n\n7-Abdel Karim M. El Hemaly, Ibrahim M Kandil, and Bahaa E. El Mohamady. Menopause, and Voiding troubles. \nhttp://www.obgyn.net/displayppt.asp?page=/English/pubs/features/presentations/El-Hemaly03/el-hemaly03-ss\n\n8-El Hemaly AKMA, Mousa L.A. Micturition and Urinary\tContinence. Int J Gynecol Obstet 1996; 42: 291-2. \n\n9-Abdel Karim M. El Hemaly.\n Urinary incontinence in gynecology, a review article.\nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/abs-urinary_incotinence_gyn_ehemaly \n\n10-El Hemaly AKMA. Nocturnal Enuresis: Pathogenesis and Treatment. \nInt Urogynecol J Pelvic Floor Dysfunct 1998;9: 129-31.\n \n11-El Hemaly AKMA, Mousa L.A.E. Stress Urinary Incontinence, a New Concept. Eur J Obstet Gynecol Reprod Biol 1996; 68: 129-35. \n\n12- El Hemaly AKMA, Kandil I. M. Stress Urinary Incontinence SUI facts and fiction. Is SUI a puzzle?! http://www.obgyn.net/displayppt.asp?page=/English/pubs/features/presentations/El-Hemaly/el-hemaly-ss\n\n13-Abdel Karim El Hemaly, Nabil Abdel Maksoud, Laila A. Mousa, Ibrahim M. Kandil, Asem Anwar, M.A.K El Hemaly and Bahaa E. El Mohamady. \nEvidence based Facts on the Pathogenesis and Management of SUI. http://www.obgyn.net/displayppt.asp?page=/English/pubs/features/presentations/El-Hemaly02/el-hemaly02-ss\n\n14- Abdel Karim M. El Hemaly*, Ibrahim M. Kandil, Mohamad A. Rizk and Mohamad A.K.M.El Hemaly.\n Urethro-plasty, a Novel Operation based on a New Concept, for the Treatment of Stress Urinary Incontinence, S.U.I., Detrusor Instability, D.I., and Mixed-type of Urinary Incontinence.\nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/urethro-plasty_01\n\n15-Ibrahim M. Kandil, Abdel Karim M. El Hemaly, Mohamad M. Radwan: Ultrasonic Assessment of the Internal Urethral Sphincter in Stress Urinary Incontinence. The Internet Journal of Gynecology and Obstetrics. 2003. Volume 2 Number 1. \n\n\n16-Abdel Karim M. El Hemaly. Nocturnal Enureses: A Novel Concept on its pathogenesis and Treatment.\nhttp://www.obgyn.net/urogynecolgy/?page=articles/nocturnal_enuresis\n\n17- Abdel Karim M. El Hemaly. Nocturnal Enureses: An Update on the pathogenesis and Treatment.\nhttp://www.obgyn.net/urogynecology/?page=/ENHLIDH/PUBD/FEATURES/\nPresentations/ Nocturnal_Enuresis/nocturnal_enuresis\n\n18-Maternal Mortality in Egypt, a cry for help and attention. The Second International Conference of the African Society of Organization & Gestosis, 1998, 3rd Annual International Conference of Ob/Gyn Department � Sohag Faculty of Medicine University. Feb. 11-13. Luxor, Egypt. \n19-Postmenopausal Osteprosis. The 2nd annual conference of Health Insurance Organization on Family Planning and its role in primary health care. Zagaziz, Egypt, February 26-27, 1997, Center of Complementary Services for Maternity and childhood care. \n20-Laparoscopic Assisted vaginal hysterectomy. 10th International Annual Congress Modern Trends in Reproductive Techniques 23-24 March 1995. Alexandria, Egypt. \n21-Immunological Studies in Pre-eclamptic Toxaemia. Proceedings of 10th Annual Ain Shams Medical Congress. Cairo, Egypt, March 6-10, 1987. \n22-Socio-demographic factorse affecting acceptability of the long-acting contraceptive injections in a rural Egyptian community. Journal of Biosocial Science 29:305, 1987. \n23-Plasma fibronectin levels hypertension during pregnancy. The Journal of the Egypt. Soc. of Ob./Gyn. 13:1, 17-21, Jan. 1987. \n24-Effect of smoking on pregnancy. Journal of Egypt. Soc. of Ob./Gyn. 12:3, 111-121, Sept 1986. \n25-Socio-demographic aspects of nausea and vomiting in early pregnancy. Journal of the Egypt. Soc. of Ob./Gyn. 12:3, 35-42, Sept. 1986. \n26-Effect of intrapartum oxygen inhalation on maternofetal blood gases and pH. Journal of the Egypt. Soc. of Ob./Gyn. 12:3, 57-64, Sept. 1986. \n27-The effect of severe pre-eclampsia on serum transaminases. The Egypt. J. Med. Sci. 7(2): 479-485, 1986. \n28-A study of placental immunoreceptors in pre-eclampsia. The Egypt. J. Med. Sci. 7(2): 211-216, 1986. \n29-Serum human placental lactogen (hpl) in normal, toxaemic and diabetic pregnant women, during pregnancy and its relation to the outcome of pregnancy. Journal of the Egypt. Soc. of Ob./Gyn. 12:2, 11-23, May 1986. \n30-Pregnancy specific B1 Glycoprotein and free estriol in the serum of normal, toxaemic and diabetic pregnant women during pregnancy and after delivery. Journal of the Egypt. Soc. of Ob./Gyn. 12:1, 63-70, Jan. 1986. Also was accepted and presented at Xith World Congress of Gynecology and Obstetrics, Berlin (West), September 15-20, 1985. \n31-Pregnancy and labor in women over the age of forty years. Accepted and presented at Al-Azhar International Medical Conference, Cairo 28-31 Dec. 1985. \n32-Effect of Copper T intra-uterine device on cervico-vaginal flora. Int. J. Gynaecol. Obstet. 23:2, 153-156, April 1985. \n33-Factors affecting the occurrence of post-Caesarean section febrile morbidity. Population Sciences, 6, 139-149, 1985. \n34-Pre-eclamptic toxaemia and its relation to H.L.A. system. Population Sciences, 6, 131-139, 1985. \n35-The menstrual pattern and occurrence of pregnancy one year after discontinuation of Depo-medroxy progesterone acetate as a postpartum contraceptive. Population Sciences, 6, 105-111, 1985. \n36-The menstrual pattern and side effects of Depo-medroxy progesterone acetate as postpartum contraceptive. Population Sciences, 6, 97-105, 1985. \n37-Actinomyces in the vaginas of women with and without intrauterine contraceptive devices. Population Sciences, 6, 77-85, 1985. \n38-Comparative efficacy of ibuprofen and etamsylate in the treatment of I.U.D. menorrhagia. Population Sciences, 6, 63-77, 1985. \n39-Changes in cervical mucus copper and zinc in women using I.U.D.�s. Population Sciences, 6, 35-41, 1985. \n40-Histochemical study of the endometrium of infertile women. Egypt. J. Histol. 8(1) 63-66, 1985. \n41-Genital flora in pre- and post-menopausal women. Egypt. J. Med. Sci. 4(2), 165-172, 1983. \n42-Evaluation of the vaginal rugae and thickness in 8 different groups. Journal of the Egypt. Soc. of Ob./Gyn. 9:2, 101-114, May 1983. \n43-The effect of menopausal status and conjugated oestrogen therapy on serum cholesterol, triglycerides and electrophoretic lipoprotein patterns. Al-Azhar Medical Journal, 12:2, 113-119, April 1983. \n44-Laparoscopic ventrosuspension: A New Technique. Int. J. Gynaecol. Obstet., 20, 129-31, 1982. \n45-The laparoscope: A useful diagnostic tool in general surgery. Al-Azhar Medical Journal, 11:4, 397-401, Oct. 1982. \n46-The value of the laparoscope in the diagnosis of polycystic ovary. Al-Azhar Medical Journal, 11:2, 153-159, April 1982. \n47-An anaesthetic approach to the management of eclampsia. Ain Shams Medical Journal, accepted for publication 1981. \n48-Laparoscopy on patients with previous lower abdominal surgery. Fertility management edited by E. Osman and M. Wahba 1981. \n49-Heart diseases with pregnancy. Population Sciences, 11, 121-130, 1981. \n50-A study of the biosocial factors affecting perinatal mortality in an Egyptian maternity hospital. Population Sciences, 6, 71-90, 1981. \n51-Pregnancy Wastage. Journal of the Egypt. Soc. of Ob./Gyn. 11:3, 57-67, Sept. 1980. \n52-Analysis of maternal deaths in Egyptian maternity hospitals. Population Sciences, 1, 59-65, 1979. \nArticles published on OBGYN.net: \n1- Abdel Karim M. El Hemaly*, Ibrahim M. Kandil, Laila A. S. Mousa and Mohamad A.K.M.El Hemaly.\nUrethro-vaginoplasty, an innovated operation for the treatment of: Stress Urinary Incontinence (SUI), Detursor Overactivity (DO), Mixed Urinary Incontinence and Anterior Vaginal Wall Descent. \nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/ urethro-vaginoplasty_01\n\n2- Abdel Karim M. El Hemaly, Ibrahim M Kandil, Mohamed M. Radwan.\n Urethro-raphy a new technique for surgical management of Stress Urinary Incontinence.\nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/\nnew-tech-urethro\n\n3- Abdel Karim M. El Hemaly, Ibrahim M Kandil, Mohamad A. Rizk, Nabil Abdel Maksoud H., Mohamad M. Radwan, Khalid Z. El Shieka, Mohamad A. K. M. El Hemaly, and Ahmad T. El Saban.\nUrethro-raphy The New Operation for the treatment of stress urinary incontinence, SUI, detrusor instability, DI, and mixed-type of urinary incontinence; short and long term results. \nhttp://www.obgyn.net/urogyn/urogyn.asp?page=urogyn/articles/\nurethroraphy-09280\n\n4-Abdel Karim M. El Hemaly, Ibrahim M Kandil, and Bahaa E. El Mohamady. Menopause, and Voiding troubles. \nhttp://www.obgyn.net/displayppt.asp?page=/English/pubs/features/presentations/El-Hemaly03/el-hemaly03-ss\n\n5-El Hemaly AKMA, Mousa L.A. Micturition and Urinary\tContinence. Int J Gynecol Obstet 1996; 42: 291-2. \n\n6-Abdel Karim M. El Hemaly.\n Urinary incontinence in gynecology, a review article.\nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/abs-urinary_incotinence_gyn_ehemaly \n\n7-El Hemaly AKMA. Nocturnal Enuresis: Pathogenesis and Treatment. \nInt Urogynecol J Pelvic Floor Dysfunct 1998;9: 129-31.\n \n8-El Hemaly AKMA, Mousa L.A.E. Stress Urinary Incontinence, a New Concept. Eur J Obstet Gynecol Reprod Biol 1996; 68: 129-35. \n\n9- El Hemaly AKMA, Kandil I. M. Stress Urinary Incontinence SUI facts and fiction. Is SUI a puzzle?! http://www.obgyn.net/displayppt.asp?page=/English/pubs/features/presentations/El-Hemaly/el-hemaly-ss\n\n10-Abdel Karim El Hemaly, Nabil Abdel Maksoud, Laila A. Mousa, Ibrahim M. Kandil, Asem Anwar, M.A.K El Hemaly and Bahaa E. El Mohamady. \nEvidence based Facts on the Pathogenesis and Management of SUI. http://www.obgyn.net/displayppt.asp?page=/English/pubs/features/presentations/El-Hemaly02/el-hemaly02-ss\n\n11- Abdel Karim M. El Hemaly*, Ibrahim M. Kandil, Mohamad A. Rizk and Mohamad A.K.M.El Hemaly.\n Urethro-plasty, a Novel Operation based on a New Concept, for the Treatment of Stress Urinary Incontinence, S.U.I., Detrusor Instability, D.I., and Mixed-type of Urinary Incontinence.\nhttp://www.obgyn.net/urogyn/urogyn.asp?page=/urogyn/articles/urethro-plasty_01\n\n12-Ibrahim M. Kandil, Abdel Karim M. El Hemaly, Mohamad M. Radwan: Ultrasonic Assessment of the Internal Urethral Sphincter in Stress Urinary Incontinence. The Internet Journal of Gynecology and Obstetrics. 2003. Volume 2 Number 1. \n\n13-Abdel Karim M. El Hemaly. Nocturnal Enureses: A Novel Concept on its pathogenesis and Treatment.\nhttp://www.obgyn.net/urogynecolgy/?page=articles/nocturnal_enuresis\n\n14- Abdel Karim M. El Hemaly. Nocturnal Enureses: An Update on the pathogenesis and Treatment.\nhttp://www.obgyn.net/urogynecology/?page=/ENHLIDH/PUBD/FEATURES/\nPresentations/ Nocturnal_Enuresis/nocturnal_enuresis",institutionString:null,institution:{name:"Al Azhar University",country:{name:"Egypt"}}},{id:"113313",title:"Dr.",name:"Abdel-Aal",middleName:null,surname:"Mantawy",slug:"abdel-aal-mantawy",fullName:"Abdel-Aal Mantawy",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Ain Shams University",country:{name:"Egypt"}}}],filtersByRegion:[{group:"region",caption:"North America",value:1,count:5681},{group:"region",caption:"Middle and South America",value:2,count:5161},{group:"region",caption:"Africa",value:3,count:1683},{group:"region",caption:"Asia",value:4,count:10200},{group:"region",caption:"Australia and 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