Dimethyl acetamide and dimethyl sulfoxide associated at glucose and egg yolk for cryopreservation of Pseudoplatystoma corruscans semen

This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.(AU)

Este estudo teve como objetivo desenvolver um protocolo para a criopreservação do sêmen de Pseudoplatystoma corruscans. Para tal, machos maduros foram induzidos hormonalmente com uma dose única de extrato de hipófise de carpa (5 mg/kg de peso vivo). O sêmen foi coletado e avaliado. Sendo testados para compor os diluentes, dois crioprotetores: dimetil acetamida (DMA) e dimetil sulfóxido (Me2SO), em duas concentrações (8% e 10%), + 5,0% glicose + 10% gema de ovo. O sêmen foi diluído na proporção 1: 4 (sêmen: extensor), embalado em palhetas de 0,5 mL e congelado em container dryshipper em vapores de nitrogênio líquido. Após o descongelamento, foram avaliados os aspectos cinéticos espermáticos, a morfologia espermática e a integridade do DNA dos espermatozoides criopreservados. Os machos de P. corruscans produziram sêmen com motilidade espermática > 80%. Todos os tratamentos proporcionaram após o descongelamento sêmen com motilidade espermática total > 40%, sem diferença significativa (P < 0,05) entre eles, como também entre os demais parâmetros cinéticos espermáticos avaliados. Os tratamentos com DMA proporcionaram uma menor fragmentação do DNA dos gametas. Malformações espermáticas foram identificadas, tanto no sêmen fresco, como no criopreservado, sendo identificado um aumento discreto dessas malformações nos espermatozoides das amostras de sêmen descongeladas de P. corruscans.(AU)

The influence of Mycoplasma species on human and canine semen quality: a review / 亚洲男科学杂志(英文版)

Mycoplasma species (spp.) are bacteria that are difficult to detect. Currently, the polymerase chain reaction (PCR) is considered the most effective diagnostic tool to detect these microorganisms in both human and veterinary medicine. There are 13 known species of human Mycoplasma and 15 species of canine Mycoplasma. Owing to the difficulties in identifying the individual species of Mycoplasma, there is a lack of information regarding which species are saprophytic and which are pathogenic. The prevalence of the individual species is also unknown. In addition, in both humans and dogs, the results of some studies on the impact of Mycoplasma are conflicting. The presence of Mycoplasma spp. on the epithelium of reproductive tract is often associated with infertility, although they are also detected in healthy individuals. The occurrence of Mycoplasma spp. is more common in dogs (even 89%) than in humans (1.3%-4%). This is probably because the pH of a dog's genital is more conducive to the growth of Mycoplasma spp. than that of humans. Phylogenetically, human and canine Mycoplasma are related, and majority of them belong to the same taxonomic group. Furthermore, 40% of canine Mycoplasma spp. are placed in common clusters with those of human. This suggests that species from the same cluster can play a similar role in the canine and human reproductive tracts. This review summarizes the current state of knowledge about the impact of Mycoplasma on canine and human male fertility as well as the prospects of further development in this field.

Application and Prospect of RNA Profiling Analysis in Forensic Body Fluid Identification / 法医学杂志

In forensic physical evidence identification, the accurate identification of the individual origin and their body fluid composition of the biological samples obtained from the crime scene play a critical role in determining the nature of a crime. In recent years, RNA profiling has become one of the fastest developing methods for body fluids identification. Due to the characteristics of tissue or body fluid specific expression, various types of RNA markers have been proven to be promising candidate markers for body fluids identification in previous studies. This review summarizes the research progress of RNA markers in body fluids identification, including the RNA markers that have been effectively verified in current research and their advantages and disadvantages. Meanwhile, this review prospects the application of RNA markers in forensic medicine.

Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification / 法医学杂志

OBJECTIVES@#To establish a system for simultaneous detection of miR-888 and miR-891a by droplet digital PCR (ddPCR), and to evaluate its application value in semen identification.@*METHODS@#The hydrolysis probes with different fluorescence modified reporter groups were designed to realize the detection of miR-888 and miR-891a by duplex ddPCR. A total of 75 samples of 5 body fluids (including peripheral blood, menstrual blood, semen, saliva and vaginal secretion) were detected. The difference analysis was conducted by Mann-Whitney U test. The semen differentiation ability of miR-888 and miR-891a was evaluated by ROC curve analysis and the optimal cut-off value was obtained.@*RESULTS@#There was no significant difference between the dual-plex assay and the single assay in this system. The detection sensitivity was up to 0.1 ng total RNA, and the intra- and inter-batch coefficients of variation were less than 15%. The expression levels of miR-888 and miR-891a detected by duplex ddPCR in semen were both higher than those in other body fluids. ROC curve analysis showed that the AUC of miR-888 was 0.976, the optimal cut-off value was 2.250 copies/μL, and the discrimination accuracy was 97.33%; the AUC of miR-891a was 1.000, the optimal cut-off value was 1.100 copies/μL, and the discrimination accuracy was 100%.@*CONCLUSIONS@#In this study, a method for detection of miR-888 and miR-891a by duplex ddPCR was successfully established. The system has good stability and repeatability and can be used for semen identification. Both miR-888 and miR-891a have high ability to identify semen, and the discrimination accuracy of miR-891a is higher.

Aflatoxins and fungal community distribution during harvesting and processing of Platycladi Semen / 中国中药杂志

This study aimed to analyze aflatoxins content and fungal community distribution in the harvesting and processing of Platycladi Semen, and explore the key link that affects aflatoxins contamination. The related Platycladi Semen samples of different maturity periods(cone non-rupture period, early rupture, and complete rupture period) and different processing periods(before drying, during 2-d drying, during 7-d drying, before and after seed scale removal, before and after peeling, 1 d after color sorting, and 7 d after color sorting) were collected for identifying the fungal community composition on sample surface by ITS amplicon sequencing. Then the content of aflatoxins B_1, B_2, G_1 and G_2 was determined by HPLC-MS/MS. The results showed that during the harvesting of Platycladi Semen from cone non-rupture to complete rupture, aflatoxins were only detected in the seed scale and seed coat, with aflatoxin G_2 in the seed scale and aflatoxin B_1 in the seed coat. During the drying, with the prolongation of drying time, aflatoxins B_1 and G_2 were detected simultaneously in the seed scale, aflatoxin B_1 in the seed coat, and low-content aflatoxin B_1 in the seed kernel. During subsequent processing, the aflatoxin content in seed kernel during subsequent processing was slighted increased. As demonstrated by fungal detection, Aspergillus flavus was not present during the harvesting of Platycladi Semen, but present during the drying and processing. Its content in the seed coat during the drying process was relatively higher. In short, Platycladi Semen should be harvested as soon as possible after it becomes fully mature. Drying process is the key link of preventing aflatoxin contamination. It is advised to build a sunlight room or adopt similar settings, standardize the operations in other processes, and keep the surrounding environment clean to minimize aflatoxin contamination.

Literature-based analysis of conversion of components in fermentation process of Sojae Semen Praeparatum / 中国中药杂志

Based on the systematic retrieval and the reported components of Sojae Semen Nigrum and Sojae Semen Praeparatum, this study conducted in-depth analysis of conversion of components in the fermentation process, and discussed types and possible mec-hanisms of conversion of chemical components, so as to provide the basis for studying technology, medicinal ingredients and quality standards. According to the analysis, there is a certain degree of conversion of nutrients(like protein, sugar, lipid), bioactive substances(like isoflavones, saponins, γ-aminobutyric acid) and other substances(like nucleosides, melanoids, biamines, etc) in the process of fermentation.

Investigation and research on fungal diversity of Platycladi Semen on Chinese market / 中国中药杂志

In the process of harvesting, production and processing, storage, and transportation, the traditional Chinese medicine Platycladi Semen is prone to mildew due to its own and environmental factors, which can nourish the production of toxic or pathogenic fungi, and even produce mycotoxins, which affects the safety of clinical medication. The 2020 edition of Chinese Pharmacopoeia limits the highest standard of aflatoxin content in Platycladi Semen. However, there are few studies on the fungal contamination of Platycladi Semen, and it is difficult to prevent and control it in a targeted manner. Therefore, based on the Illumina NovaSeq6000 platform, this article uses ITS sequence amplicon technology to analyze the distribution and diversity of fungi in 27 batches of commercially available Platycladi Semen in the Chinese market. A total of 10 phyla, 35 classes, 93 orders, 193 families, 336 genera, and 372 species of fungi were identified in China. Among them, Aspergillus, Alternaria spp. were dominant, 20 batches of samples were detected for A. flavus, 10 batches of samples were detected for A. nidulans, and all samples were detected for potential pathogenic fungi such as A. fumigatus and A. niger. According to diversity analysis, the diversity of the fungal communities in the samples from Gansu province was high, the samples in Shandong province contain the largest number of fungal species, and the samples in Guangxi province had the lo-west diversity and the least number of species. In most samples, pathogenic fungi such as A. fumigatus, A. niger, A. flavus, A. parasiticus were detected in varying degrees. This study systematically investigated the fungal contamination of Platycladi Semen from the markets in the last link of the its industrial chain, and clarified the distribution of Platycladi Semen fungi, especially toxin-producing fungi, and provided theoretical basis for the targeted prevention and control of fungal contamination in Platycladi Semen.

High-speed identification of odor changes and substance basis of Myristicae Semen mildew by electronic nose and HS-GC-MS / 中国中药杂志

This study adopted headspace-gas chromatography-mass spectrometry(HS-GC-MS) and electronic nose to detect volatile components from Myristicae Semen samples with varying degrees of mildew, aiming at rapidly identifying odor changes and substance basis of Myristicae Semen mildew. The experimental data were analyzed by electronic nose and principal component analysis(PCA). The results showed that Myristicae Semen samples were divided into the following three categories by electronic nose and PCA: mildew-free samples, slightly mildewy samples, and mildewy samples. Myristicae Semen samples with different degrees of mildew greatly varied in volatile components. The volatile components in the samples were qualitatively and quantitatively detected by HS-GC-MS, and 59 compounds were obtained. There were significant differences in the composition and content in Myristicae Semen samples with different degrees of mildew. The PCA results were the same as those by electronic nose. Among them, 3-crene, D-limonene, and other terpenes were important indicators for the identification of mildew. Bicyclo[3.1.0]hexane, 4-methylene-1-(1-methylethyl)-, terpinen-4-ol, and other alcohols were key substances to distinguish the degree of mildew. In the later stage of mildew, Myristicae Semen produced a small amount of hydroxyl and aldehyde compounds such as acetaldehyde, 2-methyl-propionaldehyde, 2-methyl-butyraldehyde, and formic acid, which were deduced as the material basis of the mildew. The results are expected to provide a basis for the rapid identification of Myristicae Semen with different degrees of mildew, odor changes, and the substance basis of mildew.

The effects of propofol and midazolam on rat spermatogenic parameters / [Efeitos do propofol e midazolam em parâmetros espermatogênicos em ratos]

There is a need for various anesthetic agents to obtain sperm in the field of human and veterinary medicine. Propofol and midazolam are among the most preferred among these agents. The aim of this study was to determine how sperm paramaters are affected according to the anesthetic agent used. Propofol (2mg/kg) and midazolam (3,5-7,5mg/kg) were administered twice a day (morning-evening) for one week. As a result of this study, there was no statistical difference in sperm density and abnormal sperm rates (respectively P=0,673, P=0,479). Sperm motility rates are similar in the control and propofol groups, while the motility rate in the midazolam group is statistically lower. (Control group %85 - Midazolam group %68.75 - Propofol group %83.75), (P<0.05). As a result of this study, the confidence interval of propofol was higher than the other anesthetic agents used for sperm retrieval.(AU)

São necessários vários agentes anestésicos para obter espermatozoides no campo da medicina humana e veterinária. Propofol e midazolam estão entre os agentes preferidos. O objetivo deste estudo foi determinar como os parâmetros de esperma são afetados de acordo com o agente anestésico utilizado. Propofol (2mg / kg) e midazolam (3,5-7,5mg / kg) foram administrados duas vezes ao dia (manhã e noite) durante uma semana. Neste estudo, não houve diferença estatística na densidade espermática e nas taxas anormais de espermatozoides (respectivamente P = 0,673, P = 0,479). As taxas de motilidade espermática são semelhantes nos grupos controle e propofol, enquanto a taxa de motilidade no grupo midazolam é estatisticamente menor. (Grupo controle % 85 - grupo midazolam % 68,75 - grupo propofol % 83,75), (P <0,05). Neste estudo, o intervalo de confiança do propofol foi maior do que os outros agentes anestésicos utilizados na recuperação espermática.(AU)

Seminal citrate is superior to PSA for detecting clinically significant prostate cancer

ABSTRACT Purpose: To establish whether the citrate concentration in the seminal fluid ([CITRATE]) measured by means of high-resolution nuclear magnetic resonance spectroscopy (1HNMRS) is superior to the serum prostate-specific antigen (PSA) concentration in detecting of clinically significant prostate cancer (csPCa) in men with persistently elevated PSA. Materials and Methods: The group of patients consisted of 31 consecutively seen men with histological diagnosis of clinically localized csPCa. The control group consisted of 28 men under long-term follow-up (mean of 8.7 ± 3.0 years) for benign prostate hyperplasia (BPH), with persistently elevated PSA (above 4 ng/mL) and several prostate biopsies negative for cancer (mean of 2.7 ± 1.3 biopsies per control). Samples of blood and seminal fluid (by masturbation) for measurement of PSA and citrate concentration, respectively, were collected from patients and controls. Citrate concentration in the seminal fluid ([CITRATE]) was determined by means of 1HNMRS. The capacities of PSA and [CITRATE] to predict csPCa were compared by means of univariate analysis and receiver operating characteristic (ROC) curves. Results: Median [CITRATE] was significantly lower among patients with csPCa compared to controls (3.93 mM/l vs. 15.53 mM/l). There was no significant difference in mean PSA between patients and controls (9.42 ng/mL vs. 8.57 ng/mL). The accuracy of [CITRATE] for detecting csPCa was significantly superior compared to PSA (74.8% vs. 54.8%). Conclusion: Measurement of [CITRATE] by means of 1HNMRS is superior to PSA for early detection of csPCa in men with elevated PSA.

High exposure to boron in drinking water and sperm parameters in Chilean young people / Exposición elevada a boro en el agua potable y parámetros de esperma en gente joven chilena

Our aim was to describe sperm parameters in residents from Northern Chile. We evaluated in 101 volunteers (18 and 30 years old) urinary and drinking water Boron levels using Inductively Coupled Plasma Mass Spectrometry; semen parameters were measured with standardized methods. Each individual was categorized in 3 levels of exposure: low (B levels in urine 2.94 mgL-1 or tap water 3.0 mgL-1), medium (urinary B between 2.95-7.4 mgL-1 and B in tap water with 3.0-7.0 mgL-1) and high (urinary B > 7.4 mgL-1 or tap water > 7.0 mgL-1). We found no significant differences among groups by pH, sperm concentration (45.1; 48.2 and 38 million/mL), motility 1th hour (38.1; 40.0 and 45.5 %) and vitality 1th hour (88.6; 88.0 and 76.9 %) respectively. Abnormal morphology was significant different (83.3; 90 and 83 %). Young men exposed to B in drinking water present sperm variations associated with the level of exposure. Most of these changes are positive at intermediate levels of B. For the highest exposures were observed negative changes in sperm morphology, concentration, motility and vitality, all relevant parameters of fertility. Beneficial effect is observed at medium exposure, like a "U curve".

El objetivo de este trabajo fue describir los parámetros espermáticos en residentes del norte de Chile. Se evaluaron en 101 voluntarios (18 y 30 años), los niveles urinarios y de agua potable de boro, usando "Inductively Coupled Plasma Mass Spectrometry". Los parámetros del semen se midieron con métodos estandarizados. Cada individuo se clasificó en 3 niveles de exposición: bajo (niveles B en la orina 2,94 mgL-1 o agua potable 3,0 mgL-1), medio (B urinario entre 2,95-7,4 mgL-1 y B en agua de beber con 3,0- 7,0 mgL-1) y alto (B urinario >7,4 mgL-1 o agua potable > 7,0 mgL-1). No se encontraron diferencias significativas entre los grupos por pH, concentración de espermatozoides (45,1; 48,2 y 38 millones/mL), motilidad a 1 hora (38,1; 40,0 y 45,5%) y vitalidad 1 hora (88,6; 88,0 y 76,9%) respectivamente. La morfología anormal fue significativamente diferente (83,3; 90 y 83%). Los hombres jóvenes expuestos a B en el agua potable presentan variaciones espermáticas asociadas con el nivel de exposición. La mayoría de estos cambios son positivos en niveles intermedios de B. Para las exposiciones más altas se observaron cambios negativos en la morfología, concentración, motilidad y vitalidad del esperma, parámetros relevantes de la fertilidad. Un efecto beneficioso se observa en la exposición media, como una "curva U".

Comparison of MPure-12 Automatic Nucleic Acid Purification and Chelex-100 Method / 法医学杂志

<p><b>OBJECTIVES</b>To explore the forensic application value of MPure-12 automatic nucleic acid purification （MPure-12 Method） for DNA extraction by extracting and typing DNA from bloodstains and various kinds of biological samples with different DNA contents.</p><p><b>METHODS</b>Nine types of biological samples, such as bloodstains, semen stains, and saliva were collected. DNA were extracted using MPure-12 method and Chelex-100 method, followed by PCR amplification and electrophoresis for obtaining STR-profiles.</p><p><b>RESULTS</b>The samples such as hair root, chutty, butt, muscular tissue, saliva stain, bloodstain and semen stain were typed successfully by MPure-12 method. Partial alleles were lacked in the samples of saliva, and the genotyping of contact swabs was unsatisfactory. Additional, all of the bloodstains （20 μL, 15 μL, 10 μL, 5 μL, 1 μL） showed good typing results using Chelex-100 method. But the loss of alleles occurred in 1 μL blood volume by MPure-12 method.</p><p><b>CONCLUSIONS</b>MPure-12 method is suitable for DNA extraction of a certain concentration blood samples．Chelex-100 method may be better for the extraction of trace blood samples．This instrument used in nucleic acid extraction has the advantages of simplicity of operator, rapidity, high extraction efficiency, high rate of reportable STR-profiles and lower man-made pollution.</p>

Inclusion of propolis in rabbit diets and semen characteristics / Inclusão de própolis na dieta de coelhos e características do sêmen / Inclusión de propóleos en la dieta de conejos y características del semen

This experiment was carried out with the objective of evaluating different levels of powder propolis in rabbit diets and their effect on semen characteristics. A total of 36 New Zealand White male rabbits were used, randomly distributed into six groups, corresponding to six propolis levels (0, 0.25, 0.50, 0.75, 1.0 and 1.25 g propolis/kg of ration). Semen was collected twice a week, using an artificial vagina. Semen volume, progressive spermatic motility, spermatic vigor, spermatic concentration and spermatic morphology were analyzed. General linear models were used for statistical analysis. The inclusion of powder propolis in the diet increased normal spermatozoa percentage and reduced spermatozoa abnormalities. The powder propolis did not affect the progressive spermatic motility, spermatic vigor or spermatic concentration. The values were considered normal for rabbits. However, a small reduction in semen volume was observed, without any negative effect on the other semen characteristics evaluated. Thus, it is possible to observe better semen quality with the inclusion of 1.25 g powder propolis/kg in the diet for reproducer rabbits...

Este experimento foi realizado com o objetivo de avaliar a influência de diferentes níveis de própolis em pó na ração de coelhos sobre as características do sêmen. Utilizaram-se 36 coelhos machos, adultos, Nova Zelândia Brancos, divididos aleatoriamente em seis grupos, consumindo cinco níveis de própolis (0; 0,25; 0,50; 0,75; 1,0 e 1,25 g de própolis/kg de ração). Coletou-se sêmen duas vezes por semana, utilizando vagina artificial. Verificou-se o volume, a motilidade espermática progressiva, o vigor espermático, a concentração espermática e a morfologia espermática. As análises estatísticas foram realizadas utilizando os modelos lineares generalizados. A adição da própolis na ração elevou a porcentagem de espermatozóides normais e reduziu os anormais. Todavia, foi observada uma pequena redução no volume do sêmen com o aumento do nível de própolis na dieta, sem afetar as demais características do sêmen. A motilidade progressiva, vigor espermático e concentração espermática não foram influenciados pelos diferentes níveis de própolis, valores considerados normais para coelhos. Conclui-se que a melhor qualidade do sêmen de coelhos reprodutores ocorreu com a adição de 1,25 g de própolis/kg de ração...

Este experimento se llevó a cabo para evaluar la influencia de diferentes niveles de polvo de propóleos en la dieta de conejos, bajo las características del semen. Se utilizó 36 conejos machos, adultos, Nueva Zelanda Blancos, divididos al azar en seis grupos, consumiendo cinco niveles de propóleos (0; 0,25; 0,50; 0,75; 1,0 y 1,25g de propóleos/kg en el alimento). Se recogió semen dos veces a la semana, utilizando vagina artificial. Se encontró el volumen, motilidad espermática progresiva, el vigor de espermático, la concentración espermática y la morfología espermática. Los análisis estadísticos se realizaron utilizando modelos lineales generales. La adición de propóleos en la dieta aumentó el porcentaje de espermatozoides normales y redujo los anormales. Sin embargo, se ha observado una pequeña reducción en el volumen del semen con el aumento de propóleos en la dieta, sin afectar las demás características del semen. La motilidad progresiva, vigor espermático y concentración de espermatozoides no se vieron afectados por los diferentes niveles de propóleos, valores considerados normales para conejos. Se concluye que la mejor calidad del semen de conejos reproductores ocurrió con la adición de 1,25g de propóleos / kg en el alimento...

Sperm parameters and biochemical components of goat seminal plasma in the rainy and dry seasons in the Brazilian Northeast: the season's influence on the cooling of semen / Caracrterísticas espermáticas e bioquímicas do plasma seminal de caprinos nas estações chuvosa e seca do Nordeste brasileiro: influência da estação no resfriamento do sêmen

The present study aimed to verify the caprine semen characteristics during dry and rainy seasons in the Brazilian Northeast, and the influence of these seasons on cooled semen. Seminal volume, concentration, percentage of motile cells, vigor and spermatic morphology, as well as biochemical profile (fructose, citric acid, P, Ca2+, Mg, total proteins and phospholipase A2 activity) were analyzed. It was observed a reduction (P<0.05) in normal sperm morphology, fructose, citric acid, P, Mg and total protein concentration during the dry season, which did not affect the motility, vigor, volume and sperm concentration. Phospholipase A2 activity was increased during the dry season (P<0.05). The analysis of the semen cooled at 4ºC during 48 hours showed reduction in total motility and vigor sperm during the dry season (P<0.05). Based on these results, we conclude that the best period of year for caprine semen cooling is the rainy season.

Verificou-se as características seminais de caprinos durante a época seca e a chuvosa no Nordeste brasileiro e a influência da época no resfriamento do sêmen. Foram mensurados volume, concentração espermática, porcentagem de espermatozoides móveis, vigor, morfologia espermática e características bioquímicas (frutose, ácido cítrico, fósforo, magnésio, proteínas totais e atividade da fosfolipase A2). Observou-se redução (P<0,05) no número de espermatozóides morfologicamente normais, frutose, ácido cítrico, fósforo, magnésio e proteínas totais durante a época seca que não influenciaram na motilidade, vigor, volume e concentração do sêmen. Entretanto, a atividade da fosfolipase A2 foi maior na época seca. Quando o sêmen foi submetido ao resfriamento a 4ºC durante 48 horas, houve redução (P<0,05) na motilidade total e no vigor espermático durante a época seca. Com base nesses resultados, conclui-se que o período chuvoso é melhor para resfriar sêmen de caprinos no Nordeste brasileiro.

Messenger RNA profiling for forensic body fluid identification: research and applications / 法医学杂志

Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts. However, the conventional body fluid identification methods are prone to various limitations, such as time consumption, intensive labor, nonparallel manner, varying degrees of sensitivity and limited specificity. Recently, the analysis of cell-specific messenger RNA expression (mRNA profiling) has been proposed to supplant conventional methods for body fluid identification. Since 2011, the collaborative exercises have been organized by the European DNA Profiling Group (EDNAP) in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification. The major advantages of mRNA profiling, compared to the conventional methods, include higher sensitivity, greater specificity, the ability of detecting several body fluids in one multiplex reaction, and compatibility with current DNA extraction and analysis procedure. In the current review, we provided an overview of the present knowledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possible practical application to forensic casework.

Influences of cycle index and volume on sensitivity of DNA detection / 法医学杂志

OBJECTIVE@#To explore the feasibility of improving the sensitivity of DNA detection by increasing the PCR cycle index and decreasing the volume of amplifying system.@*METHODS@#The DNA of semen were collected from 10 healthy irrelevant volunteers, and were quantified to 50, 40, 30, 25, 20, 15, 10 pg/microL, separately. All samples were then amplified in 10, 5, 3 microL volume and at 28, 30, 32, 34, 36 cycles, respectively. 3130 genetic analyzer was used to detect 15 autosomal STR loci.@*RESULTS@#Under the situation of 28 cycles and 3 microL volume, samples which achieved > 40 pg/microL could be correctly typed. Under the situation of 10, 5, 3 microL volume, samples which achieved > 20 pg/microL could be correctly typed at 34 cycles. When increasing the index to 36 cycles, they could not be correctly typed because of the non-specific band.@*CONCLUSION@#DNA detecting sensitivity can be improved to a certain extent by increasing the cycle index and decreasing the volume of amplifying system.

Apoptotic markers in semen of infertile men: association with cigarette smoking

OBJECTIVES: (i) To examine the role of apoptosis in the pathogenesis of DNA damage in semen from infertile men. (ii) To assess the effects of smoking on apoptotic markers and seminal parameters among infertile men. (iii) To assess the correlation of apoptosis with conventional semen parameters. MATERIALS AND METHODS: The study was carried out on 70 men with idiopathic infertility, divided into two groups: thirty infertile non smokers and forty infertile smokers. In addition to 60 fertile men (30 non smokers and 30 smokers) as control group. Each subject provided semen for analysis of parameters, determination of percent of DNA fragmentation, s-Fas, caspase-3 activity levels and cotinine levels. RESULTS: The results revealed that infertile men, particularly smokers have significantly lower semen variables and significantly higher levels of apoptotic variables ( percent of DNA fragmentation, s-Fas and caspase-3 activity) in addition to cotinine. CONCLUSIONS: The present findings provide additional evidence supporting the importance of the evaluation of apoptotic markers to test male infertility particularly among smokers.

Evaluation of testosterone hormone and zinc Levels among infertile males in Kirkuk province/ Iraq

Male Infertility is often caused by problems with sperm production or motility. Zinc in human semen seems to play an important role in the physiology of spermatozoa This study was designed to demonstrate the relationships between concentrations of zinc and testosterone in serum and seminal plasma and sperm quality among infertile men. One hundred four infertile males, aged [19-44] years, were selected from Infertile Clinic-Azadi Teaching Hospital- Kirkuk Province. Forty known fertile males were selected as normospermic control group. Semen samples were analyzed according to WHO criteria. Serum and seminal plasma zinc concentrations were estimated by atomic absorption technique. Serum testosterone was measured by MiniVIDAS apparatus. The mean value of serum testosterone was significantly lower in infertile males [4.87 +/- 0.15 ng/ml] as compared to control group [6.41 +/- 0.16 ng/ml]; [P< 0.01], significant correlations were observed between serum testosterone with seminal plasma zinc level in oligospermic subjects [r=0.44] and with serum zinc level in azoospermic subjects [r=0.37], [P< 0.01]; [P< 0.05] respectively. Serum and seminal plasma zinc levels were lower in infertile men [7.75 +/- 0.18 micromol/L]; [0.83 +/- 0.02 mmol/L] when compared with normospermic control group [14.09 +/- 0.27 Mmol/L]; [1.41 +/- 0.01 mmol/L] respectively [p<0.01], Zinc may contribute to fertility through its positive effect on spermatogenesis. Also there was significant decrease in serum and seminal plasma zinc levels in oligospermic and azoospermic infertile males with significantly low androgen. It indicates that the zinc may have a role for steroidogenesis

Seminal fructose in various classes of infertile patients

A great number of substances have been found in sperm plasma but so far it has not been possible to provide evidence of clinical significance for all of them. Fructose occupies the most important place on biochemical investigations. Fructose acts as a donor of energy to the spermatozoa. Fructose is secreted from the seminal vesicles and the accessory sex glands. It is the major carbohydrate found in seminal plasma, and appears essential for normal sperm motility. We present results of a prospective study of seminal fructose in patients referred for routine semen analysis prior to infertility treatment. Qualitative measurement of fructose in seminal fluid was carried out by Resorcinol method. Fructose level in various groups of male infertility, and sperm concentration in various groups was estimated. They were classified as azoospermic, oligozoospermic, polyzoospermic, normozoospermic, asthenozoospermic, and teratozoospermic on the basis of sperm concentrations, motility and morphology respectively. It is indicated that the true corrected fructose level is a simple method for assessment of the seminal vesicular function

Influence of feed restriction on chromosomal pictures, some biochemical parameters and spermiogram of Crossbred rams

The goal of the present study is to evaluate the effect of feed restriction on chromosomal pictures, serum biochemical parameters and some hormones in addition to spermiogram with their reflection on the reproductive efficiency of rams. Five mature Crossbred [Barki x Rahmani] rams were used, their age ranged from 15 to 17 months and weighting 45 to 60 kg. Rams received a basal diet according to the management of Animal Reproduction Research Institute [ARRI]. The animals were exposed to 21 days of feed restriction [1/3 amount of basal diet]. Restricted rams refed again on basal diet for 14 days. Blood and semen samples were collected from rams [before feed restriction, 21 days after feed restriction and 14 days post refeeding]. The results showed that there were significant differences between first period and others in the percentage of some chromosomal abnormalities which mainly were polydiploidy, in addition to gaps, breaks, deletions and fragments which increased significantly only after feed restricted periods in addition to the significant increase in the percent of total number of aberrated cells. Concerning the biochemical parameters, there was a significant decrease in serum glucose and cholesterol levels after feed restriction and they were elevated after refeeding to their levels before the restriction. Malondialdehyde showed a significant decrease after refeeding than the other two periods of the experiment. On the contrary, there was a significant increase in serum insulin levels after feed restriction compared with the other two periods. The effect of feed restriction was clear on serum testosterone hormone level which showed significant decrease after feed restriction and after refeeding. There was significant decrease in semen volume after restriction and refeeding rams. There was significant decrease in the motility and live/dead sperm after feed restriction and return to their levels post refeeding. Concentration of semen showed significant decrease after both feed restriction and refeeding. Total sperm abnormalities showed significant increase after restriction and back to first level before restriction. It was concluded that, feed restriction [l/3 amount of basal ration] for 21 days in mature rams has adverse effect on the chromosomal pictures, some biochemical parameters and decreased testosterone level in addition to decrease in all semen parameters. This effect was not overcome by supplemental feeding again that confirming the status of these rams did not improve completely even after the tested refeeding period