Sararat Innoi

Morus alba L. fruit extract (MFE), which is rich in cyanidin 3-glucoside (C3G), demonstrates antioxidant properties and pharmacological effects, but its reproductive safety remains poorly understood. Polyphenols modulate steroidogenesis, spermatogenesis, and sperm acrosome integrity; therefore, toxicity assessment is needed for use. This study aimed to evaluate the antioxidant profiles and subchronic reproductive effects of MFE. MFE was standardized using high-performance liquid chromatography (HPLC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. Male rats were administered MFE (250 or 500 mg/kg BW) for 56 days. Assessments included computer-assisted sperm analysis (CASA), testosterone, seminal fructosamine, and testicular CYP11A1 and androgen receptor (AR) expression. Acrosomal status was determined using PNA lectin staining. The results showed that MFE contained C3G (119.42 mg/g), antioxidant capacity (DPPH IC₅₀: 0.101 mg/mL; FRAP: 465.01 µmol Fe (II)/g), and total contents (phenolics: 41.15 mg GAE/g; flavonoids: 3.15 mg CE/g; anthocyanidins: 11.04 mg C3G/g). MFE did not alter testicular histology and seminiferous stages VII-IX. High doses significantly increased sperm concentration, while both doses reduced sperm beat cross frequency. Testosterone, fructosamine, and CYP11A1/AR expressions showed increasing trends. Significantly, high doses induced a precocious acrosome reaction. In conclusion, MFE has no reproductive toxicity and pro-fertility effects on sperm quantity or androgenic markers, supporting safe subchronic use.

Background/Objectives: Sex estimation from craniofacial morphology is a fundamental component of biological profile construction in forensic anthropology. Population-specific reference data for Thai individuals derived from computed tomography (CT) remain limited, and direct comparisons between discriminant function analysis (DFA) and machine learning classifiers are frequently complicated by inconsistent validation protocols. This study aimed to characterize sexual dimorphism in CT-derived craniofacial measurements, compare the classification performance of DFA, support vector machine (SVM), and random forest (RF) under a unified validation protocol, and demonstrate their practical application in a forensic context. Methods: CT images from 300 Thai adults (150 males, 150 females; age range 20–90 years) were obtained from Srinagarind Hospital, Khon Kaen University. Eight linear craniofacial measurements spanning the cranial vault, facial skeleton, nasal aperture, and orbital region were obtained from each case. DFA, SVM, and RF were developed and compared under a unified leave-one-out cross-validation protocol. Classification performance was assessed using accuracy, AUC, and Matthews correlation coefficient (MCC). Results: Seven of eight measurements exhibited statistically significant sexual dimorphism, with facial breadth and nasal height demonstrating the greatest dimorphism. DFA achieved the highest classification accuracy of 85.7%, AUC of 0.924, and MCC of 0.713, incorporating five measurements into the canonical function. SVM and RF achieved comparable accuracy of 84.7% and 84.0%, respectively. All three classifiers correctly classified both forensic application cases with high confidence. Conclusions: CT-derived craniofacial measurements provide a reliable basis for sex estimation in Thai adults. The convergence of performance across all three classifiers under a unified internal validation protocol strengthens confidence in the internally validated performance estimates. The derived discriminant function equation and saved machine learning models constitute a complementary and immediately applicable toolkit for CT-based forensic sex estimation in the Thai population.

Chronic stress (CS) contributes to male infertility, reduced testosterone levels, and impaired semen quality. CS models induced by glucocorticoids, such as dexamethasone (DEX), negatively affect sperm parameters and testicular health, notably by promoting testicular apoptosis. While individual plant extracts have been studied for their ability to mitigate stress-induced reproductive dysfunction, the preventive effect of the Tri Garn Pis (TGP) polyherbal extract in DEX-induced CS (DexCS) has not previously been investigated. This study evaluated the effects of TGP extract on testicular function, sexual behavior, and sperm quality in DexCS male mice. Seventy-two ICR mice were randomly divided into six groups: control, DexCS, TGP (50, 100, and 200) + DexCS, and TGP200. Mice received TGP (50, 100, 200 mg/kgBW) for 14 days before DEX co-treatment for 28 days. Behavioral and reproductive assessments included depression-like behavior tests, sexual behavior, sperm quality, testicular histopathology, steroidogenesis proteins (AR, CYP11A1, StAR), and apoptosis markers (Hsp70, caspase-3, caspase-9). TGP extract—which is rich in phenolics and flavonoids with antioxidant activity—improved depressive behavior, sexual performance, testicular histology, and low sperm quality. TGP also upregulated testicular StAR expression while reducing caspase-3 and caspase-9 levels. TGP prevents testicular apoptosis, sexual dysfunction, and poor sperm motility induced by DexCS.

Dexamethasone (DEX), a synthetic glucocorticoid widely prescribed for chronic inflammatory conditions, autoimmune disorders, and acute pain management, has been implicated in male reproductive dysfunction. Although many studies have documented DEX-induced oxidative stress, apoptosis, and testosterone reduction in rodent models, the molecular mechanisms underlying steroidogenic disruption and tyrosine phosphorylation remain incompletely understood. Aim of this study was to investigate the effects of chronic DEX administration on key steroidogenic proteins and tyrosine phosphorylated (TyrPho) protein profiles in rat testis. Adult rats received either DEX (1.5 mg/kg body weight, intraperitoneally) or vehicle control daily for 21 consecutive days. Results showed that DEX treatment significantly reduced testicular weight, seminiferous tubule diameter, and epithelial height compared to those of control. Tubular atrophy and interstitial tissue degeneration were observed in DEX-treated testes. Western blot analysis demonstrated significant downregulation of steroidogenic acute regulatory protein (StAR) and cytochrome P450 family 11 subfamily A member 1 (CYP11A1), while heat shock protein 70 (HSP70) expression was not different. Immunofluorescence confirmed the reduced StAR localization in Leydig cells of DEX group. TyrPho protein profiling revealed differential responses, with DEX significantly increasing a 70 kDa protein band while decreasing a 25 kDa protein band; other Tyrpho proteins (50, 42, and 38 kDa, respectively) not affected. It was concluded that chronic DEX exposure compromised the testicular architecture and suppressed critical steroidogenic machinery, providing mechanistic insights into glucocorticoid-induced hypogonadism and reproductive toxicity in male mammals.

Background/Objectives: Determination of the biological profile, particularly sex and stature, constitutes an essential component for individual identification in forensic and archaeological anthropology; however, validation of anthropometric equations remains inadequately implemented in contemporary research. This study addresses two limitations: the isolated development of sex and stature estimation methods and the lack of rigorous validation using independent samples. Methods: In this research, we analyzed 400 well-preserved scapulae from a Northeastern Thai population divided into training (n = 300: 150 male scapulae, 150 female scapulae) and validation (n = 100: 50 male scapulae, 50 female scapulae) groups. Eight standardized measurements were used for both sex and stature estimation. Results: All measurements demonstrated significant sexual dimorphism, with males exhibiting larger dimensions. For sex estimation, a multivariate model incorporating the maximum scapular height, maximum length of the spine, and scapula weight achieved 96.3% accuracy in the training samples and maintained 95.0% accuracy in independent validation. For stature estimation, a three-variable equation combining scapula weight, longitudinal scapular length, and maximum scapular breadth was strongly correlated (R = 0.769, SEE = 5.32 cm) with consistent performance across validation samples. Conclusions: Validation testing confirmed the high accuracy, reliability, and stability of both equations when applied to independent samples, with no significant differences in performance metrics between training and validation groups. These validated equations provide reliable standards for forensic practitioners analyzing scapular remains in practical applications within the northeastern Thai population.

Chronic stress (CS) is known to induce testicular oxidative stress and apoptosis. Fruit of Momordica charantia (FMC) has antioxidant capacity to protect tissue damage, but its effect on sexual activity and testicular damage caused by CS has never been documented. This study aimed to investigate the impact of FMC extract against testicular damage and sexual dysfunction in chronic unpredictable stress (CUS) rats. Rats were divided into four groups and pretreated with vehicle or FMC (40 and 80 mg/kg) before CUS induction for 56 days. Sexual behaviors, serum hormones, and sperm quality were analyzed. Testes were collected to determine daily sperm production (DSP), malondialdehyde (MDA) level, and expressions of cleaved apoptotic proteins. Testicular DNA fragmentation, as revealed by TUNEL and morphometric analysis, was observed. FMC improved sexual activity, increased testosterone levels, and sperm count with improvements in DSP, testicular morphometrics, and MDA levels. Moreover, TUNEL-positive cells and expression of caspase 3 in the testis were decreased in FMC rats. FMC has antioxidant potential and could protect against male sexual dysfunction and testicular damage caused by stress-related apoptosis. It is a potential extract to be developed as a supplement in preventing CS-male subfertility. However, future studies about the optimal dose and clinical trials are required.

Chronic stress (CS) from physical stressors and glucocorticoid administrations has been shown to induce germ cell apoptosis and low sperm quality. Among CS animal models, the effects of dexamethasone induced CS (DexCS) on testicular apoptosis and epididymal sperm parameters have not been fully demonstrated. This study aimed to investigate the changes of male reproductive system after CS induction by Dex. Adult male mice were divided into control and DexCS groups. Control mice were injected with sodium phosphate while DexCS mice were injected with DEX (4 mg/kg BW) for 21 consecutive days. The stress tests (sucrose preference, tail suspension, and forced swimming) were used to confirm CS behaviors. Sperm concentration, motility, morphology, viability, and acrosome status were assessed using computer-assisted semen analysis (CASA) and special staining. Histopathology of testis, epididymis, and penis was observed. Apoptotic protein expressions (Hsp70, caspases 3 and 9) in the testicular lysate were also determined. The results revealed that DexCS mice had significant increase of the immobility periods with decrease of total sucrose intake. DexCS significantly decreased sperm quality parameters particularly progressive motility. Testicular damages and decreased sperm mass in epididymis were obviously found in DexCS group. The expressions of testicular caspases 3 and 9, but not Hsp 70 were significantly increased in DexCS group compared to the control. It was concluded that DEX is a potential drug to induce chronic stress in mouse model, affecting male reproductive system via testicular histopathology and apoptotic pathway. Such effect may cause sperm physiology impairments like low progressive motile patterns.