Heat-shock protein 110 (HSP110) family members act as nucleotide exchange factors (NEF) of mammalian and yeast HSP70 chaperones during the ATP hydrolysis cycle. In this study, we describe the expression pattern of murine HSPA4, a member of the HSP110 family, during testis development and the consequence of HSPA4 deficiency on male fertility. HSPA4 is ubiquitously expressed in all the examined tissues. During prenatal and postnatal development of gonad, HSPA4 is expressed in both somatic and germ cells; however, expression was much higher in germ cells of prenatal gonads. Analyses of Hspa4-deficient mice revealed that all homozygous mice on the hybrid C57BL/6J!129/Sv genetic background were apparently healthy. Although HSPA4 is expressed as early as E13.5 in male gonad, a lack of histological differences between Hspa4 K/K and control littermates suggests that Hspa4 deficiency does not impair the gonocytes or their development to spermatogonia. Remarkably, an increased number of the Hspa4-deficient males displayed impaired fertility, whereas females were fertile. The total number of spermatozoa and their motility were drastically reduced in infertile Hspa4-deficient mice compared with wild-type littermates. The majority of pachytene spermatocytes in the juvenile Hspa4 K/K mice failed to complete the first meiotic prophase and became apoptotic. Furthermore, down-regulation of transcription levels of genes known to be expressed in spermatocytes at late stages of prophase I and post-meiotic spermatids leads to suggest that the development of most spermatogenic cells is arrested at late stages of meiotic prophase I. These results provide evidence that HSPA4 is required for normal spermatogenesis.
Aims: This study aims to evaluate the impact of solid-state fermentation (SSF) by Trichoderma reesei on the phenolic content, antioxidant and antimicrobial activities of garden cress seeds (GCS). Methods and Results: The factorial statistical design was employed to optimize the SSF conditions, incubation time, pH, temperature and moisture, for maximum production of the phenolic content and microbial carbohydratecleaving enzymes from GCS. The total phenolic content significantly increased from unfermented GCS (401 mg gallic acid equivalent (GAE) 100 g À1 ) to fermented GCS (3600 mg GAE 100 g À1 ) by ninefold. The total antioxidant activity significantly increased in fermented GCS. Fifteen phenolic compounds were detected in fermented GCS with high concentrations compared to 14 in unfermented GCS using high-performance liquid chromatography. A strong correlation between the production of the carbohydrate-cleaving enzymes and the phenolic content of fermented GCS was observed. The phenolic compounds of fermented GCS showed higher antimicrobial activity. Conclusions: The fermented GCS is a powerful source of phenolic compounds with high antioxidant potentials, which can be used as dietary supplement and antimicrobial agent. Significance and Impact of the Study: Solid-state fermentation is a promising technique used for production of added-value bioactive compounds. SSF increased the total phenolic content and antioxidant activity of GCS several folds compared to germination process, which recently studied.
Chia seeds (CS) are becoming increasingly consumed due to their great nutritional and therapeutic properties. In this study, solid-state fermentation (SSF) of CS by Trichoderma reesei was employed to maximize the production of the antioxidant-phenolic compounds and some fungal phenolic-associated enzymes (α-amylase, xylanase, β-glucosidase, polygalacturonase, and phenylalanine ammonia-lyase). The SSF-conditions were statistically optimized using response surface methodology (RSM). In the statistical model, four variables were analyzed at two levels. According to RSM, the adjusted R2 (< 0.9) is reasonably consistent with the predicted R2 (< 0.9), indicating that the statistical model is valid. The optimal conditions for maximum production of both phenolic compounds and fungal phenolic-associated enzymes were found to be 28 °C, pH 7.0, 20% moisture, and 7-day fermentation. The total phenolic content of fermented CS (FCS) increased 23 folds and total antioxidant activity was enhanced by 113- and 150-fold using DPPH and ABTS methods, respectively. Three new phenolics (kaempferol, apigenin, and p-coumaric) were recognized in FCS using HPLC analysis. The activities of all the extracted phenolic-associated enzymes showed strong correlations with the phenolic content of FCS. Against some human-pathogenic bacteria, FCS extract displayed considerably better antibacterial activity than UFCS extract. Finally, the phenolic-rich-FCS can be employed as a dietary supplement as well as an antibacterial agent. Furthermore, T. reesei has produced considerable quantities of industrially valuable enzymes.
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