Anatomy of mamillo-accessory foramen and prevalence of ossified mamillo-accessory ligament in lumbar vertebrae related to age.

PURPOSE: Ossification of the mamillo-accessory ligament (MAL) results in the formation of a mamillo-accessory foramen (MAF), which is associated with aging. The MAL tethers the medial branches of the lumbar dorsal rami to the lumbar vertebrae. A MAL ossified at the lumbar vertebrae can cause low back pain by compressing the medial branch of a dorsal ramus. Age ranges related to ossification of the MAL have not been reported in previous studies. The objective of the present study was to determine the prevalence of ossification of the MAL in the lumbar column and its relationship to aging, and to measure the newly formed MAF at each level of the lumbar vertebrae. METHODS: This study examined 935 dried lumbar vertebrae from 187 donors at Khon Kaen University, Thailand, consisting of 93 females and 94 males. The research focused on ossification patterns of the MAL, categorizing them into three patterns. RESULTS: We found that over 50% of ossified MAL occurred in the 30-45-year-old range and the frequency increased with age. The prevalence of ossified lumbar MAL was 72.73%, especially in L5 on the left side in females (76.92%). The width of the MAF did not differ significantly between the sexes, but it was greater on the left side (2.46 ± 1.08; n = 76) than the right (2.05 ± 0.95; n = 72) (p = 0.016). CONCLUSION: Ossification of the MAL into the MAF progresses with age, leading to low back pain from nerve compression. Physicians should be aware of the MAF during anesthesia block to treat low back pain.

Morphometric analysis of dry atlas vertebrae in a northeastern Thai population and possible correlation with sex.

PURPOSE: The uppermost segment of the cervical vertebra or atlas (C1) is a critically important anatomical structure, housing the medulla oblongata and containing the grooves for the C1 spinal nerve and the vertebral vessels. Variations of the C1 vertebra can affect upper spine stability, and morphometric parameters have been reported to differ by population. However, there are few data regarding these parameters in Thais. The use of this bone to predict sex and age has never been reported. METHODS: This study aimed to examine C1 morphometry and determine its ability to predict sex. Twelve diameter parameters were taken from the C1 vertebrae of identified skeletons (n = 104, males [n, 54], females [n, 50]). Correlation analysis was also performed for sex and age, which were predicted using machine learning algorithms. RESULTS: The results showed that 8 of the 12 measured parameters were significantly longer in the male atlas (p < 0.05), while the remaining 4 (distance between both medial-most edges of the transverse foramen, transverse dimension of the superior articular surface, frontal plane passing through the canal's midpoint, and anteroposterior dimension of the inferior articular surface) did not differ significantly by sex. There was no statistically significant difference in these parameters on the lateral side. The decision stump classifier was trained on C1 parameters, and the resulting model could predict sex with 82.6% accuracy (root mean square error = 0.38). CONCLUSION: Assertation of the morphometric parameters of the atlas is important for preoperative assessment, especially for the treatment of atlas dislocation. Our findings also highlighted the potential use of atlas measurements for sex prediction.

Topographical study of scapular foramina and scapular nutrient foramina in dried skeletons.

PURPOSE: The aim of our study is to study the prevalence and anatomy of scapular foramina (SF) and scapular nutrient foramina (SNF) in dried skeletons from the Northeastern Thai population. METHODS: A total of 150 dried scapulae were investigated. Both SF and SNF were identified using a metal wire with a diameter of 0.36 mm. The number, locations, lengths, and diameters of SF were recorded. Subsequently, SNF were identified using the same metal wire. Their number and locations were recorded. Two observers performed the evaluations and measurements. RESULTS: SF were present in 78.0% of scapulae. They could have up to five openings. Eighteen types were found. On average they were longer in males (21.7 ± 5.0 mm) than females (19.45 ± 4.6 mm). The mean diameters of both the superior and inferior openings were significantly greater in females (p < 0.01). SNF, in contrast, were present in 100% of scapulae. They were located in the supraspinous fossa (36.7%), subscapular fossa (31.3%), infraspinous fossa (22.8%), and peri-glenoid area (10.0%). CONCLUSION: Unlike previous studies, the present study suggests that SF are normal anatomical findings, present in 78.0% of the scapulae investigated. Surgeons should be aware of both SNF and SF when operating or interpreting radiological findings.

Anatomical characteristics and morphometric analyses of the internal carotid artery using retrospective angiographic images.

PURPOSE: Anatomical variations of the internal carotid artery (ICA) can cause complications during endovascular treatments. Therefore, the aim of this study was to investigate the features of the ICAs obtained from 2D digital subtraction angiography (2D DSA). METHODS: The morphometrics and angles of the ICA segments from 2D DSA images from a total of 100 patients (45 males and 55 females) were investigated. The lengths (L1-3), angles (A1-3), and diameters (D1-6) through the ICA measurement points (five segments: C2 [petrous], C3 [lacerum], C4 [cavernous], C5 [clinoid], C6 [ophthalmic]) were systemically recorded by two observers. All measured parameters were compared for both sexes and sides. RESULTS: The lengths (mm) and angles of the ICA were 7.20 ± 2.22 (L1), 15.71 ± 2.32 (L2), 10.99 ± 1.66 (L3) and 109.31 ± 17.77 (A1), 107.87 ± 20.51 (A2), 80.81 ± 16.33 (A3), respectively. There were no differences in the angulations of the A1-A3 segments between the sexes (p > 0.05). The L2 (C3-C4) was significantly longer in males, but only the left side of females showed a greater length (p < 0.05). The averaged mean diameters of the ICA (D1-6) in both sexes were 4.17 ± 0.55 mm. However, the diameters of the D1-D6 from left and right sides of males except the right D6 were significantly longer than those of females (p < 0.05). Also of significance was that the right diameters of females (D1) and males (D1 and D2) were shorter compared to left sides. CONCLUSION: This study demonstrates some differences in lengths, diameters, and angles in both sexes and sides of the normal ICA. Neurosurgeons and neurointerventionalists should be aware of such variations when operating, performing interventional procedures, and interpreting 2D DSA.

Prevention of seminal vesicle damage by Mucuna pruriens var. pruriens seed extract in chronic unpredictable mild stress mice.

CONTEXT: Thai Mucuna pruriens (L.) DC. var. pruriens (Fabaceae) or T-MP seed extract has been shown to improve sexual performance and sperm quality. OBJECTIVE: This study investigates the preventive effects of T-MP against seminal vesicle damage, apoptotic and Nrf2 protein expression in mice under chronic unpredictable mild stress (CUMS). MATERIALS AND METHODS: Forty-eight male ICR mice were divided into four groups: control, CUMS, T-MP300 + CUMS and T-MP600 + CUMS. Mice in control and CUMS groups received distilled water, while those in treated groups were pretreated with T-MP extract (300 or 600 mg/kg BW) for 14 consecutive days. The CMUS and co-treated groups were exposed to one random stressor (of 12 total) each day for 43 days. Components and histopathology of the seminal vesicle were examined, along with localization of androgen receptor (AR) and caspase 3. Expression of seminal AR, tyrosine phosphorylated (TyrPho), heat shock protein 70 (Hsp70), caspases (3 and 9) and nuclear factor erythroid 2-related factor 2 (Nrf2) proteins was investigated. RESULTS: T-MP extract at a dose of 600 mg/kg BW improved seminal epithelial damage and secretion of fluid containing essential substances and proteins in CUMS mice. It also increased the expression of AR and TyrPho proteins. Additionally, T-MP increased expression of Nrf2 and inhibited seminal vesicular apoptosis through the suppression of Hsp70 and caspase expression. CONCLUSION: T-MP seeds have an antiapoptotic property in chronic stress seminal vesicle. It is possible to apply this extract for the enhancement of seminal plasma quality.

Dolichandrone serrulata flower improves seminal biochemical parameters and proteins in T2DM rats induced by a high-fat diet and streptozotocin.

CONTEXT: Although Dolichandrone serrulata (Wall. ex DC.) Seem (Bignoniaceae) flower (DSF) improves hyperglycaemia, testicular damage and sperm quality in type 2 diabetes mellitus (T2DM) animals, its effects on the seminal vesicles, secreting seminal plasma, are unknown. OBJECTIVE: This study reports the protective effects of DSF on seminal dysfunction in T2DM rats. MATERIALS AND METHODS: Male Sprague-Dawley rats were divided into four groups (control, T2DM, T2DM + DSF200 and T2DM + DSF600; 10 animals/group). The control group was fed a low-fat diet for 14 days prior to single saline injection, whereas T2DM group was given a high-fat diet and injected with streptozocin (50 mg/kg body weight). The T2DM-induced rats were fed DSF orogastrically (200 and 600 mg/kg body weight) for 28 consecutive days. At the end of the experiment, biochemical components, malondialdehyde (MDA), histology and protein expression in seminal lysate were evaluated. RESULTS: DSF increased the levels of serum phosphorus (13.66 ± 0.59 mg/dL), ALP (11.85 ± 0.99 U/L), GOT (3938.23 ± 251.41 U/L) and GPT (34.16 ± 4.93), decreased MDA levels in seminal tissue, and elevated the serum testosterone in the T2DM rats. Treatment with DSF ameliorated histological damage, significantly increased seminal 44 and 31 kDa TyrPho protein expression, and decreased that of caspase 3 and 9. CONCLUSIONS: DSF extract was able to mitigate seminal dysfunction in T2DM rats via improvements of tyrosine phosphorylation, testosterone level and biochemical substances, as well as reductions of caspase proteins. DSF may be developed as an alternative medicine in treating of T2DM male subfertility and progressive complications.

Seed extract of Thai Mucuna pruriens reduced male reproductive damage in rats induced by chronic stress.

CONTEXT: Thai Mucuna pruriens (L.) DC. var. pruriens (Fabaceae) (TMP) is known to enrich reproduction but preventive effects on stress related adverse reproductive parameters are not documented. OBJECTIVE: This study investigates the protective property of TMP seed extract on reproductive damage under chronic stress (CS). MATERIALS AND METHODS: Male Sprague-Dawley rats were divided into four groups. The control and CS groups received distilled water, whereas the pre-treated rats received the aqueous TMP seed extract at doses of 150 and 300 mg/kg BW for 20 days before co-treatments with CS induction (immobilization and forced swimming) for 81 days. Serum was used to determine the cortisol and testosterone levels. Histology of testis and epididymis was observed with localization of androgen receptor (AR). Sperm parameters and the expression of steroidogenic acute regulatory (StAR), cytochrome P450 family 11 subfamily a member 1 (CYP11A1), AR, HSP70, caspases (3 and 9) and tyrosine phosphorylation (TyrPho) proteins were investigated. RESULTS: TMP extract improved cortisol level (0.84 ± 0.02 µg/dL) and protected against the damage of reproductive tissues and sperm parameters (count [49.78 ± 3.74 million sperm/mL], viability [90.01 ± 1.17%] and precocious acrosome reaction [1.38 ± 0.48%]). Expression of testicular StAR, CYP11A1, AR and HSP70 proteins was improved. Caspase expression was decreased in treated rats. TMP increased AR expression in CS sperm. Moreover, TyrPho protein expression was corrected after TMP administration. CONCLUSIONS: TMP seed protected against adverse reproductive parameters in CS via improvements of functionally testicular markers and reductions of apoptotic proteins. It is possible to develop the TMP beans as an alternative medicine in treating of male subfertility caused by CS.

Dolichandrone serrulata flower extract ameliorates male reproductive damages in type 2 diabetic rats.

Dolichandrone serrulata flower (DSF) has been believed to reduce blood glucose in hyperglycaemic persons with sub-fertility but its effect on improvement of male reproductive impairment has never been elucidated scientifically. This study attempted to investigate the hypoglycaemic effects of DSF on male reproductive damages in type 2 diabetes mellitus (T2DM) rats. Adult Sprague Dawley rats were divided into four groups (control, T2DM, DSF200 + T2DM and DSF600 + T2DM; n = 10/each). Control rats received low-fat diet for 14 days before saline injection while streptozocin (50 mg/kg BW) induced T2DM groups received high-fat diet and were orally administered with DSF (200 and 600 mg/kg BW) for 28 days. At the end, fasted blood glucose (FBG), malondialdehyde (MDA), testosterone, sperm quality, histology and protein expressions were examined. The result showed that DSF decreased high FBG and testicular MDA and increased testosterone levels of T2DM-treated rats. Low-sperm quality and histological malfunction were ameliorated in DSF-treated group. There was significant decrease in the expression of androgen receptor, heat-shock 70 and steroidogenic acute regulatory proteins of T2DM-treated rats. Our study demonstrated changes of six bands (116, 51, 45, 39, 35 and 29 kDas) of tyrosine-phosphorylated proteins. In conclusion, DSF could reduce the FBGand ameliorate the reproductive damages in male T2DM rats.

Decreased expression of AKAP4 and TyrPho proteins in testis, epididymis, and spermatozoa with low sexual performance of mice induced by modified CUMS.

The molecular mechanism of chronic stress especially reduced motility, a major cause of male infertility, has not been proved. It is known that A-kinase anchor protein 4 (AKAP4) and tyrosine-phosphorylated (TyrPho) proteins are involved in progressive motility. This study aimed to investigate the effect of chronic unpredictable mild stress (CUMS) on sexual behaviours, sperm quality, and expressions of AKAP4 and TyrPho proteins in testis, epididymis, and spermatozoa. Sixteen male mice were divided into control and CUMS groups (n = 8/group). Animals were induced by a stressor from twelve stressors for 36 days. Sexual behaviours, corticosterone and testosterone, sperm parameters, and histopathology were observed. The expressions of AKAP4 and TyrPho proteins in testis, epididymis, and spermatozoa were examined. Results showed that CUMS significantly increased corticosterone while serum testosterone level was decreased. Sexual behaviours and sperm parameter quality were significantly decreased. CUMS mice showed vacuolisation and pyknotic cells in seminiferous epithelium and less sperm mass was observed within epididymal lumen. CUMS decreased expressions of AKAP4 and TyrPho proteins in testis, epididymis, and spermatozoa. In conclusion, the decreased expression of AKAP4 and TyrPho proteins may be a mechanism associated with low semen qualities particularly decrease of sperm motility in CUMS.

Effect of chronic stress on expression and secretion of seminal vesicle proteins in adult rats.

Chronic stress (CS) is known to affect men's health especially fertility by reducing semen quality. Although the effects of CS on testicular function and sperm parameters are documented, changes of substances and secreting proteins in the seminal vesicle (SV) have never been reported. This study aimed to demonstrate the alterations of contents and expressions of proteins in seminal vesicle fluid (SVF) under CS. Fourteen adult rats were divided into control and CS groups (n = 7/each). Control rats were not exposed to stressor, while the CS animals were immobilised by restraint cage (4 hr/day) and followed by forced swimming (15 min/day) for consecutive 60 days. Biochemical substances and malondialdehyde (MDA) level in SVF were examined. Expressions of heat-shock protein 70 (Hsp70), caspases (Casp) 3 and 9, and tyrosine-phosphorylated (TyrPho) proteins were investigated in seminal vesicle tissue (SVT) and SVF. It was found that CS caused reductions of seminal epithelial height and secreted substance levels. Significantly, MDA levels in SVF and expressions of Hsp70, Casp and TyrPho proteins were increased in of CS animals. It was concluded that CS affected seminal secretion. Low quality of CS seminal plasma may associate with increase of MDA and expressions of secreted proteins.

Chronic stress affects tyrosine phosphorylated protein expression and secretion of male rat epididymis.

Chronic stress (CS) is shown to decrease the semen quality with changed expression of tyrosine phosphorylated (TyrPho) proteins in testicular and seminal tissues. However, the alterations of such proteins and fluid contents in the epididymis, producing sperm maturation factors, have never been reported. Sixteen adult rats were randomly divided into 2 groups (n = 8). The control animals were not subjected to stressors whereas CS rats were immobilised within restraint cage (4 hr/day) before cold forced-water swimming (15 min/day) for 60 days. Corticosterone, testosterone, blood glucose level (BGL), malondialdehyde (MDA) and biochemical components in epididymal fluid were assayed. Expressions of heat shock protein 70 (HSP-70), androgen receptor (AR) and TyrPho protein were investigated in epididymal tissue and fluid. Significantly, CS increased the corticosterone and BGL but decreased testosterone and epididymal substance levels. MDA level in tail epididymal fluid and HSP-70 expression in both regions of epididymal tissues and fluids, except in head epididymal fluid of CS were increased. Epididymal tissues showed the decrease of AR expression. Presence and changes of many TyrPho proteins were observed in CS. In conclusion, CS could affect functional proteins particularly TyrPho in epididymis, resulted in low semen quality.

The effect of Dolichandrone serrulata (wall. ex DC.) Seem. flower extract containing antioxidant capacity and terpenoids on the male reproductive system.

Although the fruit extract of Dolichandrone genus was shown to inhibit spermatogenesis, the reproductive toxicity of Dolichandrone serrulata flowers (DSFs) is not documented. Recent study aimed to evaluate the sub-chronic toxicity of DSF on male reproductive system. Antioxidant capacity and total phenolic contents of DSF extract were determined using Folin-Ciocalteu's, 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power assays. The terpenoid components were determined using nuclear magnetic resonance spectrum. Adult male rats were treated orally with DSF (100, 300 or 600 mg/kg) for 48 days. Histopathology of testis and epididymis was observed. Sperm concentration, viability, acrosome status and morphology were also examined. Expressions of heat shock protein 70 (Hsp70), tyrosine-phosphorylated (TyrPho) proteins, androgen receptor (AR) and steroidogenic acute regulatory (StAR) protein in testis were investigated. Results showed that DSF contained phenolic compounds and terpenoids (phytoandrogens; rengyolone and cleroindicin B). No reproductive histopathology was observed in DSF-treated rats. Although DSF decreased the serum testosterone level, the sperm qualities were not affected. Particularly, sperm concentration of DSF-treated animals was significantly increased. DSF changed the testicular TyrPho proteins but the expression of AR, StAR or Hsp70 was not altered. In conclusion, DSF possesses antioxidant capacity with no toxicity on male reproductive system.

Classification and Morphometric Features of Pterion in Thai Population with Potential Sex Prediction.

Background and Objectives: The landmark for neurosurgical approaches to access brain lesion is the pterion. The aim of the present study is to classify and examine the prevalence of all types of pterion variations and perform morphometric measurements from previously defined anthropological landmarks. Materials and methods: One-hundred and twenty-four Thai dried skulls were investigated. Classification and morphometric measurement of the pterion was performed. Machine learning models were also used to interpret the morphometric findings with respect to sex and age estimation. Results: Spheno-parietal type was the most common type (62.1%), followed by epipteric (11.7%), fronto-temporal (5.2%) and stellate (1.2%). Complete synostosis of the pterion suture was present in 18.5% and was only present in males. While most morphometric measurements were similar between males and females, the distances from the pterion center to the mastoid process and to the external occipital protuberance were longer in males. Random forest algorithm could predict sex with 80.7% accuracy (root mean square error = 0.38) when the pterion morphometric data were provided. Correlational analysis indicated that the distances from the pterion center to the anterior aspect of the frontozygomatic suture and to the zygomatic angle were positively correlated with age, which may serve as basis for age estimation in the future. Conclusions: Further studies are needed to explore the use of machine learning in anatomical studies and morphometry-based sex and age estimation. Thorough understanding of the anatomy of the pterion is clinically useful when planning pterional craniotomy, particularly when the position of the pterion may change with age.

Ovatodiolide, isolated from Anisomeles indica, suppresses bladder carcinogenesis through suppression of mTOR/ß-catenin/CDK6 and exosomal miR-21 derived from M2 tumor-associated macrophages.

Bladder cancer (BCa) is the fourth leading cause of cancer deaths worldwide due to its aggressiveness and resistance against therapies. Intricate interactions between cancer cells and the tumor microenvironment (TME) are essential for both disease progression and regression. Thus, interrupting molecular communications within the TME could potentially provide improved therapeutic efficacies. M2-polarized tumor-associated macrophages (M2 TAMs) were shown to contribute to BCa progression and drug resistance. We attempted to provide evidence for ovatodiolide (OV) as a potential therapeutic agent that targets both TME and BCa cells. First, tumor-suppressing functions of OV were determined by cell viability, colony, and tumor-sphere formation assays using a coculture system composed of M2 TAMs/BCa cells. Subsequently, we demonstrated that extracellular vesicles (EVs) isolated from M2 TAMs containing oncomiR-21 and mRNAs, including Akt, STAT3, mTOR, and ß-catenin, promoted cisplatin (CDDP) resistance, migration, and tumor-sphere generation in BCa cells, through increasing CDK6, mTOR, STAT3, and ß-catenin expression. OV treatment also prevented M2 polarization and reduced EV cargos from M2 TAMs. Finally, in vivo data demonstrated that OV treatment overcame CDDP resistance. OV only and the OV + CDDP combination both resulted in significant reductions in mTOR, ß-catenin, CDK6, and miR-21 expression in tumor samples and EVs isolated from serum. Collectively, we demonstrated that M2 TAMs induced malignant properties in BCa cells, in part via oncogenic EVs. OV treatment prevented M2 TAM polarization, reduced EV cargos derived from M2 TAMs, and suppressed ß-catenin/mTOR/CDK6 signaling. These findings provide preclinical evidence for OV as a single or adjuvant agent for treating drug-resistant BCa.

Effect of ethanol on the changes in testicular protein expression in adult male rats.

Ethanol consumption is a major cause of male infertility, but the exact mechanism is still largely unknown. This study attempted to investigate the effect of ethanol on sperm morphology, acrosome reaction status and the alteration of the testicular protein expressions. Fourteen male rats were divided into control and ethanol groups (n = 7/each group). Ethanol-treated rats received ethanol (5 g/kg, 40% v/v) via oral gavage for consecutive 14 days. Testosterone hormone, sperm parameters, and testicular and epididymal histopathologies were evaluated. In addition, the expressions of testicular proteins including androgen receptor (AR), heat shock protein 70 (HSP70), steroidogenic acute regulatory protein (StAR) and tyrosine-phosphorylated (TyrPho) proteins were investigated. The results showed that ethanol significantly increased percentage of abnormal sperm morphology and acrosome-reacted spermatozoa. Some seminiferous and ductus epididymal histopathologies were observed in ethanol-treated rats. Significantly, ethanol reduced serum testosterone and expressions of testicular AR and TyrPho proteins. However, the overexpression of StAR and HSP70 proteins in ethanol testis was found. It was concluded that the changes in testicular protein expressions may be involved in mechanism of male infertility caused from ethanol consumption.

Alterations of morphology and phosphorylated protein expression in the seminal vesicles of diabetic mice.

Although many studies reported the detrimental effects of type 1 and 2 diabetes mellitus (T1DM and T2DM) on testis, reproductive parameter changes in DM seminal vesicles have never been documented. This study aimed to examine the morphology, biochemical levels and tyrosine phosphorylation in seminal vesicles of T1DM and T2DM mice. Fifty-six male C57BL/6 mice were divided into four groups (n = 14/each): T1DM control, T1DM, T2DM control and T2DM. T1DM mice were daily injected of streptozotocin (STZ; 40 mg/kg BW) for 5 days. T2DM mice received high-fat diet for 14 days prior to STZ injection at a single dose (85 mg/kg BW). At the end of experiments (days 36 and 72), magnesium (MG) and fructosamine (FRA) levels, and phosphorylated protein expression in seminal vesicle were examined. The results showed that seminal and prostate weights and MG and FRA levels of T1DM animals were significantly increased as compared to T2DM mice. Some seminal histopathologies and decreased epithelial height were observed in both DM groups. Significantly, a 72-kDa phosphorylated protein expression was increased in DM seminal vesicle. We concluded that changes of biochemical components and phosphorylated proteins in seminal vesicle of T1DM and T2DM mice may be associated with low-quality seminal plasma.

Valproic acid changes the expression of tyrosine-phosphorylated proteins in rat seminal vesicle.

Previous studies reported the effects of valproic acid (VPA) on tyrosine-phosphorylated (TyrPho) protein expression in the testis and epididymis, but its effects on that in the seminal vesicle (SV) have never been demonstrated. This study attempted to investigate the expressions of TyrPho proteins in SV treated with VPA. Sixteen rats were divided into control and VPA-treated groups. The control rats were injected with normal saline, whereas the treated animals were intraperitoneally injected with VPA (500 mg/kg BW) for 10 consecutive days (n = 8/each). The biochemical parameters in blood plasma and SV fluid were analysed. The SV tissues and fluid were investigated for the presence and expression of TyrPho proteins, androgen receptor (AR) proteins and heat shock protein 70 (Hsp70). Significantly, VPA caused SV atrophy and reductions in secretion and biochemical parameter levels. There were significant increases in many TyrPho proteins in the plasma (a 95 kDa) and SV tissue (a 40 kDa) of the VPA rats. However, the expressions of seminal AR, Hsp70 and TyrPho proteins (50 and 48 kDa) were significantly lower in VPA rats. Recent results have indicated that VPA affected SV morphology and decreased biochemical fluid substances including TyrPho proteins associated with decreased expressions of AR and Hsp70.

Changes in testicular function proteins and sperm acrosome status in rats treated with valproic acid.

Valproic acid (VPA), an anti-epileptic drug, reduces testosterone levels and sperm quality. However, the degree to which testosterone levels and sperm quality are decreased under VPA treatment needs to be clarified. The aim of the present study was to investigate the testicular proteins involved in testosterone synthesis and spermatogenesis, histopathology and sperm acrosome status in VPA-treated rats. Adult rats were divided into control and experimental groups (n=8 in each). Rats in the experimental group were treated with 500mg kg-1, i.p., VPA for 10 consecutive days. Expression of Ki-67, tyrosine phosphorylated proteins and testicular steroidogenic proteins was examined. As expected, VPA-treated rats exhibited adverse changes in almost all reproductive parameters, particularly an increase in precocious acrosome reactions, compared with the control group. In addition, fibrosis of the tunica albuginea and tubule basement membrane was observed in testes from VPA-treated rats. Moreover, the expression of testicular Ki-67, cholesterol side-chain cleavage enzyme (P450scc) and phosphorylated proteins (41, 51 and 83 kDa) was decreased significantly in VPA-treated rats compared with control. In contrast, the expression of steroidogenic acute regulatory proteins in the VPA-treated group was significantly higher than in the control group. In conclusion, VPA treatment changes the expression of testicular proteins responsible for spermatogenesis and testosterone production, resulting in male infertility.

Immunohistochemical analysis of sustentacular cells in the adrenal medulla, carotid body and sympathetic ganglion of mice using an antibody against brain-type fatty acid binding protein (B-FABP).

Little attention has been paid to adrenal sustentacular cells, and several major histology textbooks do not even describe them. This study presents a detailed morphological description of sustentacular cells using immuno-light microscopy and an antibody against brain-type fatty acid-binding protein. The immunopositive sustentacular cells and processes formed lattices with holes of various sizes and compactnesses or openness. In addition, weakly immunostained sheet-like structures with ill-defined contours were often associated with the processes and lattices. In the carotid body, which has traditionally been classified under the name of paraganglia in common with the adrenal medulla, immunostained sustentacular cell processes formed lattices in association with the weakly immunostained sheet-like structures, but the lattices with sheets were more compact and rigid than the adrenal medulla, and appeared like individually distinct compartments. In the ganglion, the immunostained satellite cell processes with the sheets tightly enclosed individual neurons. As a result, the immunostained sheet-like structures were regarded as en-face views of thinly flattened sustentacular cytoplasmic envelopes partially covering the chromaffin cells in the adrenal medulla, and widely in the carotid body in a way rather similar to the satellite cells in the ganglion. In brief, the terminal enclosing portions of adrenal sustentacular cell processes, in cut-views, were too thin/flat to be recognized as distinct lines in immuno-light microscopy because of its resolution limit. They are recognized in en-face views as entities of a substantially spacious extension in immuno-light microscopy.

Rats Orally Administered with Ethyl Alcohol for a Prolonged Time Show Histopathology of the Epididymis and Seminal Vesicle Together with Changes in the Luminal Metabolite Composition.

Prolonged ethanol (EtOH) consumption is associated with male infertility, with a decreased spermatogenesis rate as one cause. The defective maturation and development of sperm during their storage in the cauda epididymis and transit in the seminal vesicle can be another cause, possibly occurring before the drastic spermatogenesis disruption. Herein, we demonstrated that the cauda epididymis and seminal vesicle of rats, orally administered with EtOH under a regimen in which spermatogenesis was still ongoing, showed histological damage, including lesions, a decreased height of the epithelial cells and increased collagen fibers in the muscle layer, which implicated fibrosis. Lipid peroxidation (shown by malondialdehyde (MDA) levels) was observed, indicating that reactive oxygen species (ROS) were produced along with acetaldehyde during EtOH metabolism by CYP2E1. MDA, acetaldehyde and other lipid peroxidation products could further damage cellular components of the cauda epididymis and seminal vesicle, and this was supported by increased apoptosis (shown by a TUNEL assay and caspase 9/caspase 3 expression) in these two tissues of EtOH-treated rats. Consequently, the functionality of the cauda epididymis and seminal vesicle in EtOH-treated rats was impaired, as demonstrated by a decreases in 1H NMR-analyzed metabolites (e.g., carnitine, fructose), which were important for sperm development, metabolism and survival in their lumen.