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Coffee fruit skin contains antioxidant compounds that can repair damaged tissues, especially those of reproductive or-gans. This study was designed to assess to what extent the metha-nolic extract of coffee fruit skin affected sperm characteristics and testicular histology in mice receiving 15% ethanol for 15 days. It used 25 male mice aged four months and employed a completely randomized design with two controls (K) and three treatments (P), namely K1 (without ethanol), K2 (15% ethanol), P1 (15% ethanol and the methanolic extract of coffee fruit skin, or MECS, at 125 mg/ kg BW), P2 (15% ethanol and MECS 250 mg/kg BW), and P3 (15% ethanol and MECS 500 mg/kg BW). These treatments were administered orally with a 1 mL disposable syringe for 15 days. The parameters observed were sperm characteristics (viability, morphology, and sperm count) and the microscopic structure of the testicles (lu-men area, diameter, and area of seminiferous tubules). These data were then analyzed using the one-way ANOVA, continued with the LSD and Duncan’s tests (P

Background and aims: Edible fiber isolated from the tuber of yam bean (Pachyrhizus erosus, Leguminosae) has been suggested to prevent the development of metabolic diseases caused by excessively consuming sugary foods. However, it is unclarified whether P. erosus fiber (PEF) is also capable of preventing liver diseases. This study aimed to determine the effectivity of PEF in counteracting the development of non-alcoholic fatty liver disease (NAFLD) caused by excessive intake of high-sugar diet (HSD). Material and method: 25% of PEF in HSD (w/w) was fed in BALB/c mice for eight weeks followed by subsequent morphological and histological examinations of the liver at the end of treatment in comparing with HSD alone. Results: We found that supplementation of 25% PEF sustained the normal morphological feature of the liver in HSD-fed mice. Moreover, PEF also prevented the development of microvesicles in the liver tissue, nuclear shrinkage, and hepatolysis as indicators of liver disease. Macrophages infiltration as a marker of liver inflammation was also less observed in PEF-treated mice. Conclusions: PEF could effectively elicit a beneficial effect against NAFLD caused by HSD. Hence, PEF is suggested to be used as a potent supplemental diet to preclude the metabolic diseases caused by HSD.

AIM: Jicama (Pachyrhizus erosus, Fabaceae) has been promoted as a potent tuberous plant exerting both preventive and therapeutic effects against many diseases. In this present study, we aimed to investigate the pancreoprotective effect of isolated jicama fiber (JF) against glucotoxicity caused by high-sugar diet (HSD). METHODS: We performed an animal experimental research using adult male BALB/c mice with a completely randomized design consisted of four treatments and six replications. The treatments were diet types such as normal diet and HSD, supplemented with 10% and 25% of JF, respectively. The treatments were deployed for 8 weeks. Furthermore, random and fasting blood glucose levels were measured; histopathological alterations of pancreas including pancreatic adiposity, size of islet area, total cell number per islet, and necrosis level of pancreatic acinar cells were examined. Quantitative data were analyzed using an ANOVA followed by Duncan’s new multiple range test (p < 0.05). RESULTS: Our results demonstrate that supplementation of JF 10% and 25% improved blood glucose profile and significantly reduced inter pancreatic adiposity caused by HSD. Moreover, JF at the dose of 25% effectively sustained normal size of the islet of Langerhans and total cell number of the islet in pancreas of HSD-fed mice. JF 25% was also effective to counteract necrosis of pancreatic acinar cells. CONCLUSION: Our findings clarify a protective effect of JF, particularly at a dose of 25%, on pancreatic tissues against degeneration caused by HSD. Hence, JF is reliable to be a supplement counteracting the development of diabetes mellitus and associated diseases.

Laminin, a major basement membrane protein, comprises three subunit chains: α, β, and γ chains. Among these chains, only the laminin α chain is capable of signaling via laminin receptors. Although laminin isoforms containing the α5 chain were reported to be the first laminin produced during rat anterior pituitary gland development, the functions of these isoforms are unknown. We used immunohistochemical techniques to localize the laminin α5 chain and its specific receptor, basal cell adhesion molecule (BCAM), in fetal and adult pituitary gland. Laminin α5 chain immunoreactivity was observed in the basement membrane of the primordial adenohypophysis at embryonic days 12.5 to 19.5. Double immunostaining showed that BCAM was present and co-localized with the laminin α5 chain in the tissue. Quantitative analysis showed that the laminin α5 chain and BCAM were expressed in the anterior pituitary gland during postnatal development and in adulthood (postnatal day 60). In the adult gland, co-localization of the laminin α5 chain and BCAM was observed, and BCAM was detected in both the folliculo-stellate cells and endothelial cells. These results suggest that laminin α5 chain signaling via BCAM occurs in both the fetal adenohypophysis and adult anterior pituitary gland.

Type 2 diabetes mellitus (T2DM) is a complex metabolic disease characterized by high blood glucose levels due to impaired insulin secretion or insulin resistance. Polymorphisms in the ABCC8 gene rs1799854 are widely found to have an association with T2DM, where the ABCC8 gene encodes the SUR1 protein from the K-ATP channel that plays a role in insulin secretion in cells β pancreas. Mutations in the ABCC8 gene can cause potassium channels and insulin secretions problems that possibly decrease the effectiveness of the drug. Polymorphisms in some populations have been reported, but similar research on Solo's population has never been conducted. The purpose of this study is to identify the genotype of the ABCC8 gene in rs1799854 and the therapeutic outcome of blood glucose levels in T2DM patients. This research is cross-sectional research conducted prospectively at Moewardi Hospital Surakarta, Solo. Blood samples were collected from 10 T2DM patients who received sulfonylurea monotherapy taken through veins for genotype examination by DNA isolation, PCR amplification, and sequencing. Parameters of fasting blood glucose (FBG), 2-hour postprandial blood glucose, and HbA1c are measured as therapeutic outcomes . We found the dominant results in a mutant homozygote (TT) 40%; while wild type (CC); and a mutant heterozygote (CT) was 30%. Single Nucleotide Polymorphisms (SNP) of ABCC8 gene rs1799854, found in T2DM patients at Moewardi Hospital Surakarta, Solo, who received sulfonylurea therapy.

The World Health Organization (WHO) has announced that Severe Acute Respiratory Syndrome Coronavirus‐2 (SARS‐CoV‐2) and Coronavirus disease (COVID‐19) is considered a worldwide pandemic. Rapidly rising numbers of patients have been reported in almost every country, along with the growing mortality rates. Uncontrolled growth in patient numbers may be due to reasons such as treatment options and vaccine availabilities and unidentified targets of SARS‐CoV‐2. Previous study has revealed that the molecular target of SARS‐CoV‐2 is analogous to SARS (2003), i.e. angiotensin‐converting enzyme‐2 (ACE‐2). Therefore, the determination of ACE‐2 may enrich existing information and facilitate development of drugs targeted toward SARS‐CoV‐2. This study aims to screen the expression of ACE‐2 genes and their relationship to the types of SNP variants in SARS‐CoV‐2. We explored a series of observations using powerful databases, e.g. GTEx portal, HaploReg, 1000 Genome and Ensembl, to identify the gene variant of ACE‐2. We showed that ACE‐2 is highly expressed in the testes and small intestine, while its lowest level is observed in lymphocytes. Subsequently, we observed 17 gene variants containing a missense mutation potentially damaging protein level. Among these genes, single nucleotide polymorphism (SNP) rs370187012 shows the highest damage‐level score, while the lowest effect is in SNP rs4646116. The highest frequency of the C allele was observed in European populations (1%). In addition to showing that ACE‐2 is expressed in several organs, we concluded that the ACE‐2 gene variation can be found in African, American, Southeast and East Asian, and European populations. The polymorphisms of ACE‐2 impact on the ACE2 protein structure and the binding capacity of the ACE‐2 receptor with the S‐Protein of SARS‐CoV‐2.

Jicama (Pachyrhizuz erosus, Fabaceae) has been reported to elicit various medicinal benefits against metabolic disturbances. We also have previously demonstrated that extracted jicama fiber (JF) could sustain normoglycemic state in high-sugar diet fed mice. In this present study, we investigated the effectivity of jicama fiber in preventing the dysregulation of energy metabolism as well as adiposity in adult male BALB/c mice fed with a high-fat diet (HFD). Three groups of animals were treated with normal diet (ND), HFD, and HFD in combination with 25% JF (w/w), respectively for 8 weeks ad libitum. Furthermore, food intake, energy intake, water intake, white adipose (WAT) and brown adipose (BAT) tissue mass, kidney and liver weight as well as organ index were determined. Our investigation revealed that JF 25% could preclude the perturbation of circadian feeding and energy intake rhythms caused by HFD but significantly reduced total water intake. JF also effectively counteracted the marked increase of WAT and decrease of BAT weight and its index in HFD fed mice. Furthermore, JF did not significantly alter the weight and index of both kidney and liver in HFD fed mice. This finding suggests that JF could be used as a potent supplement to minimize the disruption of energy homeostasis and obesity caused by HFD.

Retinoic acid (RA) is converted from retinal by retinaldehyde dehydrogenases (RALDHs) and is an essential signaling molecule in embryonic and adult tissue. We previously reported that RALDH1 was produced in the rat anterior pituitary gland and hypothesized that RA was generated in the gland. Midkine (MK) is an RA-inducible growth factor, and MK production in the rat anterior pituitary gland was recently reported. However, the mechanism that regulates gene expression of MK in the pituitary gland has not been determined. To investigate regulation of MK production in the anterior pituitary gland, we analyzed changes in MK mRNA in cultured rat anterior pituitary cells. We identified MK-expressing cells by double-staining with in situ hybridization and immunohistochemical techniques for RALDH1. MK mRNA was expressed in RALDH1-producing cells in the anterior pituitary gland. Using isolated anterior pituitary cells of rats, we examined the effect of RA on gene expression of MK. Quantitative real-time PCR revealed that 72 h exposure to a concentration of 10-6 M of retinal and all-trans retinoic acid increased MK mRNA levels by about 2-fold. Moreover, the stimulatory effect of all-trans retinoic acid was mimicked by the RA receptor agonist Am80. This is the first report to show that RA is important in regulating MK expression in rat anterior pituitary gland.

Midkine (MK) belongs to a family of secreted heparin-binding growth factors and is highly expressed in various tissues during development. MK has multiple functions, such as regulation of cell proliferation, migration, survival and differentiation. We recently reported that MK mRNA is strongly expressed in the developing rat pituitary gland. In the adult pituitary, however, expression of MK and its receptor and the characteristics of the cells that produce them, have not been determined. Therefore, in this study, we investigate whether MK and its receptor, protein tyrosine phosphatase receptor-type Z (Ptprz1), are present in the adult rat pituitary. In situ hybridization, real-time reverse transcription-PCR and immunoblotting were performed to assess MK and Ptprz1 expression. We also characterize MK- and Ptprz1-expressing cells by double-staining with in situ hybridization and immunohistochemical techniques for each pituitary hormone or S100 protein [a marker of folliculostellate (FS) cells]. MK-expressing cells were located in the anterior and posterior lobes but not in the intermediate lobe. Double-staining and immunoblotting revealed that MK mRNA and protein were only expressed in FS cells in the anterior pituitary. Regarding Ptprz1 expression, Ptprz1 mRNA was detected in adrenocorticotropic hormone (ACTH) cells and growth hormone (GH) cells but not in prolactin cells, thyroid-stimulating hormone cells, luteinizing hormone cells, or FS cells. These findings suggest that MK produced in FS cells acts locally on ACTH cells and GH cells via Ptprz1 in the adult rat anterior pituitary.

Pituitary gland development is controlled by numerous signaling molecules, which are produced in the oral ectoderm and diencephalon. A newly described family of heparin-binding growth factors, namely midkine (MK)/pleiotrophin (PTN), is involved in regulating the growth and differentiation of many tissues and organs. Using in situ hybridization with digoxigenin-labeled cRNA probes, we detected cells expressing MK and PTN in the developing rat pituitary gland. At embryonic day 12.5 (E12.5), MK expression was localized in Rathke's pouch (derived from the oral ectoderm) and in the neurohypophyseal bud (derived from the diencephalon). From E12.5 to E19.5, MK mRNA was expressed in the developing neurohypophysis, and expression gradually decreased in the developing adenohypophysis. To characterize MK-expressing cells, we performed double-staining of MK mRNA and anterior pituitary hormones. At E19.5, no MK-expressing cells were stained with any hormone. In contrast, PTN was expressed only in the neurohypophysis primordium during all embryonic stages. In situ hybridization clearly showed that MK was expressed in primitive (immature/undifferentiated) adenohypophyseal cells and neurohypophyseal cells, whereas PTN was expressed only in neurohypophyseal cells. Thus, MK and PTN might play roles as signaling molecules during pituitary development.

The anterior pituitary gland is organized tissue comprising hormone-producing cells and folliculostellate (FS) cells. FS cells interconnect to form a meshwork, and their cytoplasmic processes are anchored by a basement membrane containing laminin. Recently, we developed a three-dimensional (3D) cell culture that reproduces this FS cell architecture. In this study of the novel function of FS cells, we used transgenic rats that express green fluorescent protein in FS cells for the 3D culture. Anterior pituitary cells were cultured with different proportions of FS cells (0%, 5%, 10%, and 20%). Anterior pituitary cells containing 5-20% FS cells formed round/oval cell aggregates, whereas amorphous cell aggregates were formed in the absence of FS cells. Interestingly, immunohistochemistry showed laminin-immunopositive cells instead of extracellular laminin deposition in FS cell-deficient cell aggregates. Double-immunostaining revealed that these laminin-immunopositive cells were gonadotrophs. Laminin mRNA expression did not differ in relation to the presence or absence of FS cells. When anterior pituitary cells with no FS cells were cultured with FS cell-conditioned medium, the proportion of laminin-immunopositive cells was lower than in control. These results suggest that a humoral factor from FS cells is required for laminin release from gonadotrophs.

The extracellular matrix (ECM) is important in creating cellular environments in tissues. Recent studies have demonstrated that ECM components are localized in anterior pituitary cells and affect cell activity. Thus, clarifying the mechanism responsible for ECM maintenance would improve understanding of gland function. Tissue inhibitors of metalloproteinases (TIMPs) are endogenous inhibitors of matrix metalloproteinases and participate in ECM degradation. In this study, we investigated whether cells expressing TIMPs are present in rat anterior pituitary gland. Reverse transcription polymerase chain reaction was used to analyze expression of the TIMP family (TIMP1-4), and cells producing TIMPs in the gland were identified by using in situ hybridization. Expression of TIMP1, TIMP2, and TIMP3 mRNAs was detected, and the TIMP-expressing cells were located in the gland. The TIMP-expressing cells were also investigated by means of double-staining with in situ hybridization and immunohistochemical techniques. Double-staining revealed that TIMP1 mRNA was expressed in folliculostellate cells. TIMP2 mRNA was detected in folliculostellate cells, prolactin cells, and thyroid-stimulating hormone cells. TIMP3 mRNA was identified in endothelial cells, pericytes, novel desmin-immunopositive perivascular cells, and folliculostellate cells. These findings indicate that TIMP1-, TIMP2-, and TIMP3-expressing cells are present in rat anterior pituitary gland and that they are involved in maintaining ECM components.