Background: In suckling piglets, transmissible gastroenteritis virus (TGEV) causes lethal diarrhea accompanied by high infection and mortality rates, leading to considerable economic losses. This study explored methods of preventing or inhibiting their production. Bovine antimicrobial peptide-13 (APB-13) has antibacterial, antiviral, and immune functions. Objectives: This study analyzed the efficacy of APB-13 against TGEV through in vivo and in vitro experiments. Methods: The effects of APB-13 toxicity and virus inhibition rate on swine testicular (ST) cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT). The impact of APB-13 on virus replication was examined through the 50% tissue culture infective dose (TCID50). The mRNA and protein levels were investigated by real-time quantitative polymerase chain reaction and western blot (WB). Tissue sections were used to detect intestinal morphological development. Results: The safe and effective concentration range of APB-13 on ST cells ranged from 0 to 62.5 ㎍/mL, and the highest viral inhibitory rate of APB-13 was 74.1%. The log10TCID50 of 62.5 ㎍/mL APB-13 was 3.63 lower than that of the virus control. The mRNA and protein expression at 62.5 ㎍/mL APB-13 was significantly lower than that of the virus control at 24 hpi. Piglets in the APB-13 group showed significantly lower viral shedding than that in the virus control group, and the pathological tissue sections of the jejunum morphology revealed significant differences between the groups. Conclusions: APB-13 exhibited good antiviral effects on TGEV in vivo and in vitro.
In this study, the production of transgenic embryo was attempted by microinjection or round spermatid cultured with foreign DNA. At first, the expression of haploid spermatids specific gene, mTP1 in mouse and hPrm2 in hamster spermatids were investigated by RT-PCR method in testes of young mice and hamster testis. The specific gene expression first appeared at 18 days post partum (dpp) in mice spermatid and 20 dpp in hamster spermatid. Therefore, the round spermatids isolated from 17 dpp mice and 19 dpp hamster were used for the introduction of foreign EGFP gene into haploid round spermatids. For the introduction of EGFP gene haploid round spermatids suspended in medium including EGFP gene were treated with a different electric field strength at 0.11, 0.18 and 0.44 ㎸/cm. After electrical stimulation, viability of testicular sperm cells and 67.6%, 66.4% and 49.9%, in mice and 62.6%, 57.9% and 27% in hamster, respectively. These values were significantly lower than those of non-treated control groups 80.5% in mouse and 69.1% in hamster After 72 hrs culture, the highest expression rate of EGFP gene, 28.5% in mice and 32.1% in hamster were obtained from tile spermatogenic cells electroporated by the field strength or 0.18 ㎸/cm. Then, the ability of fertilization and embryonic development of haploid spermatids transfected with foreign EGFP gene were estimated by the microinjection of spermatids into hamster oocytes. The Irate pronuclear formation rate (77.5%) was lower than non-treated control (80%), and the cleavage rate of the treated group (58.8%) was lower than control (65%). To prove the foreign EGFP integration in hamster embryos, 2-cell stage hamster embryos were subjected to the observation under the fluorescence microscope, and the PCR analysis. As a result, about 44% of 2-cell embryos were showed the integration of EFGP gene into their genome. Therefore, These results suggest the possibility to produce transgenic hamsters by microinjection of haploid spermatid transfected with foreign DNA.
The sesarmid crab Muradium tetragonum, inhabiting the mangrove, are considered as a key consumer of litter and thereby play an important role in the detritus food chain and energy flow in the mangrove ecosystem. The present investigation was carried out with objectives to enlighten the reproductive system of Muradium tetragonum through histological and histochemical studies. Histological organization of the testis of M. tetragonum revealed that each testis has a lobular structure consisting of several testicular lobules arranged around the collecting duct. Histology of the deferens of M. tetragonum revealed it to be composed of three-layer of tissues along the entire length:the outer connective tissue, the middle muscular and the inner epithelial layer. Based on the histological architecture these three regions are recognized as proximal vas deferens (PVD), middle vas deferens (MVD) and distal vas deferens (DVD). Histological characteristics of the ovary of M. tetragonum during different phases of ovarian development were studied. Based on the colour changes of the ovary and diameter of the oocytes five stages of ovarian development can be pronounced. Histochemical analysis of the male reproductive tissues of M. tetragonum signifies the secretion of a different biomolecule by specifying their origin in the reproductive tissue and their possible transformation into spermatophores. In the female reproductive tissues, histochemical evaluation envisaged the secretory products during different stages of ovarian development The secretory substances of the spermatheca expound on the significance of its secretion in dehiscing the spermatophore wall and in nourishing as well as protecting the spermatozoa.
Nathan L. DeBono;Robert D. Daniels ;Laura E. Beane Freeman ;Judith M. Graber ;Johnni Hansen ;Lauren R. Teras ;Tim Driscoll ;Kristina Kjaerheim;Paul A. Demers ;Deborah C. Glass;David Kriebel;Tracy L. Kirkham;Roland Wedekind;Adalberto M. Filho;Leslie Stayner ;Mary K. Schubauer-Berigan
Safety and Health at Work
/
v.14
no.2
/
pp.141-152
/
2023
Objective: We performed a meta-analysis of epidemiological results for the association between occupational exposure as a firefighter and cancer as part of the broader evidence synthesis work of the IARC Monographs program. Methods: A systematic literature search was conducted to identify cohort studies of firefighters followed for cancer incidence and mortality. Studies were evaluated for the influence of key biases on results. Random-effects meta-analysis models were used to estimate the association between ever-employment and duration of employment as a firefighter and risk of 12 selected cancers. The impact of bias was explored in sensitivity analyses. Results: Among the 16 included cancer incidence studies, the estimated meta-rate ratio, 95% confidence interval (CI), and heterogeneity statistic (I2) for ever-employment as a career firefighter compared mostly to general populations were 1.58 (1.14-2.20, 8%) for mesothelioma, 1.16 (1.08-1.26, 0%) for bladder cancer, 1.21 (1.12-1.32, 81%) for prostate cancer, 1.37 (1.03-1.82, 56%) for testicular cancer, 1.19 (1.07-1.32, 37%) for colon cancer, 1.36 (1.15-1.62, 83%) for melanoma, 1.12 (1.01-1.25, 0%) for non-Hodgkin lymphoma, 1.28 (1.02-1.61, 40%) for thyroid cancer, and 1.09 (0.92-1.29, 55%) for kidney cancer. Ever-employment as a firefighter was not positively associated with lung, nervous system, or stomach cancer. Results for mesothelioma and bladder cancer exhibited low heterogeneity and were largely robust across sensitivity analyses. Conclusions: There is epidemiological evidence to support a causal relationship between occupational exposure as a firefighter and certain cancers. Challenges persist in the body of evidence related to the quality of exposure assessment, confounding, and medical surveillance bias.
Seong Jin Kim;Hyeon Jin Kim;So Ryung Shin;Myeong Gyo Seo;Pyeong Woo Kim;Eun Ha Kim;Jung Sick Lee
Journal of Marine Life Science
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v.8
no.2
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pp.178-185
/
2023
This study was described the microscopic anatomy of male reproductive organs and spermatophore necessary for understanding the reproductive ecology of the long arm octopus Octopus minor. The long arm octopus was a species that has sexual dimorphism that can distinguish between sex based on the presence of hectocotylus. Male reproductive organs consisted of testis, primary spermatic duct, spermatic gland, secondary spermatic duct, spermatophoric gland and spermatophoric sac. Histologically, the testis was testicular tubule type and male germ cells showed a layered arrangement. The primary spermatic duct was a tube connecting the testis and spermatic gland, and consisted with epithelial layer and connective tissue. The spermatic gland was located between the primary and secondary spermatic duct, and the epithelial layer was composed of epithelial cells and mucous cells. Mucous cells reacted blue in the AB-PAS (pH 2.5) reaction and purple in the AF-AB (pH 2.5) reaction. The secondary spermatic duct was a short tube connecting spermatic gland and spermatophoric gland, and folds were developed in lumen. The spermatophoric gland consisted of numerous tubular glands and secretory cells had eosinophilic granules. The spermatophoric sac was shape of pouch, folds were developed in lumen, and vacuolar secretory cells were present in the epithelial layer. The spermatophore was 83.5 mm long and consisted of cap thread in anterior portion, ejaculatory apparatus and cement body in medial portion, sperm mass in posterior portion.
Formerly, adult-tiger puffer, Takifugu rubripes with ova caught in the sea, were used for seedling production. But it was difficult to secure naturally-ripened adults. For the purpose of adult tiger puffer in captivity, this study was carried out. To determine the growth 220 tiger puffers hatched in 1990 (3-year-old) and 1991 (2-year-old) were used. For spawning and egg incubation leading to fry development, eggs were stripped from tiger puffers hatched in 1988 (5-year-old) and 1990 (3-year-old) through human chorionic gonadotropin (BCG) treatments. In May, 1993, mean body length and mean body weight of 2-year-old tiger puffer were $30.72\pm1.35cm\;and\;1,048\pm228 g,$ and that of 3-year-old tiger puffers were $36.02\pm1.17cm$ and $1,402\pm66g$ respectively. The relationship between body length (L) and body weight (W) of 2-year-old the tiger puffers during the experiment period was represented as $W\;=\;1.7892L^{31524}\times10^5$ (r= 0.9436) and that of 3-year-old, $W=\;3.2840L^{36099}\times10^6$ (r= 0.9070) respectively. The GSI in female 2-year-old-fish changed from $0.23\times0.l2\;to\;0.74\pm0.08$, during the experiment period, and in male it didn't change remarkably until November, but thereafter it increased and showed a peak of $8.69\pm5.09$. The GSI of 3-year-old-fish showed a peak of $8.05\pm5.58$ in April in female and $12.65\pm4.60$ in May in male. The change of HSI in 3-year-old-fish was correlative to the change of GSI, but in 2-year-old-fish it was little correlative. In female gonad of 2-year-old tiger puffer, the mature oocytes reached $350{\mu}m$ in April, but thereafter they didn't spawn and became atrophied. But in male gonad, a great number of spermatozoa were crowded in the testicular lobuli in April. Female gonad of 3-year-old tiger puffer had the mature oocytes of 650 pm in March and the ripe oocytes, $900{\mu}m$ in April. Male testis development was similar to that of 2-year-old-fish. Egg-stripping after hormone treatments was possible past 139 hours and 142 hours from each of two 5-year-old-fish (500IU/kg, BW), and after 114 hour from a 3-year-old-fish (1,000 IU/kg, BW) under water temperature $16.3\~17.8^{\circ}C$. Eggs stripped amounted was 650 g and 400 g from two 5-year-old-fish and 610 g from the 3-year-old-fish, and fertilization rates were $98.0\%,\;97.4\%\;and\;96.5\%$ respectively. All the hatched larvae devloped into normal fry.
Most of endocrine disrupters (EDs) have been reported to exhibit estrogenic or anti-androgenic activity and thereby may disrupt reproductive development in human or wildlife. This study was performed to investigate the effects of estrogen (E$_2$), bisphenol (BP) and octylphenol (OP) on the mouse Leydig cell line (TM3). TM3 originated from testis of 11~13-daly-old BALB/c nu/+ mice was cultured in DMEM supplemented with 10% FBS alone or medium with estrogen (E$_2$), bisphenol (BP) and octylphenol (OP; 1 pM, 1 nM, 1 $\mu$M, 1 mM, respectively) for 48 hours. After culture, total cell number and viability were assessed by heamocyto-meter and trypan blue stain. Expression of cytochrome P450scc (CYPscc) mRNA whose product is involved in steroid hormone biosynthesis and estrogen receptor $\alpha$(ER $\alpha$) mRNA were detected by RT-PCR. As a result, treatment of TM3 with E$_2$, BP and OP(1 mM, respectively) significantly decreased the viability but not all of groups as high as 1 $\mu$M. Exposure of TM3 to OP significantly reduced the total cell number but not E$_2$ or BP. The expression of CYPscc mRNA was slightly reduced in BP (1 nM, 1 $\mu$M) and significantly decreased in OP (1 nM, 1 $\mu$M) treated TM3, except E$_2$ group. But the expression of ER $\alpha$ mRNA was sightly increased in all treated groups. In conclusion, BP and OP (high concentration) might inhibit steroidogenesis by decreasing the CYPscc mRNA expression in the mouse testis. These results suggest that BP and OP might impair spermatogenesis and subsequently disturb testicular function.
Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.
Ethane 1,2-dimethane sulfonate (EDS) is a well-known alkylating agent used as selective Leydig cell (LC) toxicant to create a testicular dysfunction model. Previous studies including our own clearly demonstrated the dramatic weight loss of the androgen dependent accessory sex organs such as epididymis, seminal vesicle and prostate gland in this 'LC knock-out' rats. The present study was performed to evaluate the effect of EDS administration on histological changes of the epididymis, seminal vesicle and prostate in adult rats. Adult male Sprague-Dawley rats (350$\sim$400 g B.W.) were injected with a single dose of EDS (75 mg/kg, i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. Tissue weights (testis, epididymis, seminal vesicle and prostate gland) were measured. The histological changes of tissue were observed by a light microscopy using hematoxylin & eosin staining. Weights of the reproductive and accessory organs progressively declined after the EDS treatments (weeks 1, 2 and 3). After this, the decrease was stopped, then gradually returned to the normal levels. There was a partial (about 60%) recovery of the epididymis weight during weeks $6{\sim}7$. The cross section of epididymis revealed an increase in thickness of the epithelium during weeks $1{\sim}3$. In contrast, considerable reduction of epithelial thickness in seminal vesicle was observed during same period. Similarly, a reduction in thickness of prostate epithelial layer was found during weeks $1{\sim}3$, then it was back to normal thickness after week 4. Taken together, the present study demonstrated that the temporally induced androgen-deficiency by EDS treatment could result the prominent alterations in histology of the accessory sex organs. Further studies on the physiological and molecular regulation of these androgen-sensitive organs using EDS model will be helpful to understand the normal and pathological development and differentiation mechanism of these organs.
Objective: To compare the pregnancy outcomes after in vitro fertilization with intracytoplasmic sperm injection (IVF-ICSI) between obstrucvtive and non-obstrucvtive azoospermia. Methods: From January 1994 to December 2002, 524 patients with obstructive azoospermia (886 cycles) and 163 patients with non-obstructive azoospermia (277 cycles) were included in this study. Microsurgical epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE) in obstructive azoospermia and TESE in non-obstructive azoospermia were perfomed to retrieve sperm, which was used for ICSI and then fertilized embryos were transferred. The results of ICSI - fertlization rate (FR), clinical pregnancy rate (CPR), clinical abortion rate (CAR) and delivery rate (DR) - were statistically analysed in obstructive versus non-obstructive azoospermia. Results: There were no differences in the number of retrieved oocytes, injected oocytes for ICSI and oocyte maturation rate. FR was significantly higher in obstructive than non-obstructive azoospermia (71.7% vs. 61.1%, p<0.001). There was no difference in CPR per embryo transfer cycle. After pregnancy was established, however, CAR was significantly higher in non-obstructive than obstructive azoospermia (25.6% vs. 12.5%, p=0.004). DR per clinical pregnancy cycle was significantly higher in obstructive than non-obstructive azoospermia (78.0% vs. 64.4%, p=0.012). In the karyotype ananlysis of abortus, abnormal karyotypes were found in 75.0% (6/8) of obstructive and 55.6% (5/9) of non-obstructive azoospermia. Conclusion: Our data show significantly higher FR in obstructive than non-obstructive azoospermia. Though there was no differrence in CPR, CAR was significantly higher in non-obstructive than obstructive azoospermia. The abortion may be related to the abnormal karyotype of embryo, but further investigations are necessary to elucidate the cause of clinical abortion in azoospermia.
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