Effect of Intermittent Hypobaric Hypoxia Exposure on HIF-1α, VEGF, and Angiogenesis in the Healing Process of Post-Tooth Extraction Sockets in Rats.

OBJECTIVE: The aim of this study was to investigate the effect of intermittent hypobaric hypoxia (IHH) exposure on the expression of hypoxia-induced factor-1α (HIF-1α) messenger RNA (mRNA), vascular endothelial growth factor-a (VEGF-a) mRNA, and angiogenesis after tooth extraction in rats. MATERIALS AND METHODS: On 45 male Sprague-Dawley rats were performed the removal of the maxillary left first molar, and then they were randomly divided into 9 groups, namely: 4 groups that were exposed to IHH for 30 minutes every day in the Hypobaric Chamber at an altitude of 18,000 feet, with 1 time hypobaric hypoxia (HH), 3 times HH, 5 times HH, and 7 times HH; 4 normoxia groups that were terminated on days 1, 3, 5, and 7 after tooth extraction; and the 1 control group. Real-time polymerase chain reaction measured the molecular changes in the socket tissue after tooth extraction in rats to evaluate the expression of HIF-1α mRNA and VEGF mRNA. Histological changes with hematoxylin and eosin staining were noted to evaluate the amount of angiogenesis in the socket after tooth extraction. Molecular and histological parameters were calculated at the end of each experiment on days 0, 1, 3, 5, and 7 after tooth extraction, which exhibited the improvement phase of the wound-healing process. RESULTS: Increases in the expression of HIF-1α mRNA, VEGF mRNA, and angiogenesis were found in the IHH group compared with the normoxia group and the control group. The expression of HIF-1α mRNA increased significantly (p < 0.05) in the group after one time HH exposure on day 1, then decreased in the IHH group (three times HH exposure, five times HH exposure, and seven times HH exposure) approaching the control group. The expression of VEGF mRNA and angiogenesis began to increase after one time HH exposure on day 1, and increased again after three times HH exposure on day 3, then increased even more after five times HH exposure on day 5, and increased very significantly (**p < 0.05) after seven times HH exposure on day 7. It showed that repeated or intermittent exposure to HH conditions induced a protective response that made cells adapt under hypoxia conditions. CONCLUSION: IHH exposure accelerates the socket healing of post-tooth extraction, which is proven by changes in HIF-1α mRNA expression and increase in VEGF mRNA expression as stimuli for angiogenesis in post-tooth extraction sockets under hypobaric hypoxic condition, which also stimulates the formation of new blood vessels, thereby increasing blood supply and accelerating wound healing.

Comparative pathology and immunohistochemistry of Newcastle disease in domestic chicken (Gallus-gallus domesticus) and Alabio duck (Anas platyrhynchos Borneo).

Background: Newcastle disease is very pathogenic in chickens, whereas in ducks, the disease does not show any clinical symptoms. Aims: To compare the clinical symptoms features, pathological lesions, viral distribution, and apoptosis response caused by the Newcastle disease virus (NDV) in domestic chicken and Alabio duck. Methods: Forty domestic chickens and 40 Alabio ducks were separated into four groups: domestic chicken and Alabio duck treatment groups, where each was infected by NDV velogenic ducks/Aceh Besar_IND/2013/eoAC080721 in 106ELD50 dosage. The domestic chicken and Alabio duck control groups were each inoculated with Phosphate Buffer Saline. The infection route was intraorbital, 0.1 ml in volume. Symptoms were observed from day 1 until day 7 post-infection (PI). Necropsy was performed on days 1, 2, 3, 5, and 7 PI for organ collection. Results: Symptoms observed were disorders in the respiratory, gastrointestinal, and nervous systems, followed by 100% mortality in domestic chickens. In Alabio ducks, there were only depression and slight lethargy. The lesion in domestic chicken day 1 appeared by the lungs, thymus, Fabricius bursa, spleen, and kidney. On day 3 PI, lesions were also found in the heart, proventriculus, duodenum, and cecal tonsil. On days 5 and 7 PI, the trachea and brain lesions were found. In Alabio ducks, lesions were found in the lungs, thymus, spleen, and proventriculus on day 1. Afterward, on day 3, light lesions were found in the heart. On day 5, lesions were found in the trachea and brain; finally, on day 7, light lesions were only found in the thymus, spleen, and brain. Immunopositive reaction NDV in domestic chicken was highest in the proventriculus, duodenum, cecal tonsil, and lymphoreticular organs. In the Alabio duck, it was highest in the duodenum and cecal tonsil. The caspase-3 percentage in domestic chicken increased on day 3 PI; in Alabio ducks, on day 2 PI. Conclusion: Clinical symptoms and pathological lesions were faster and more severe in domestic chickens. The immunopositive reaction NDV in domestic chicken continued to increase, while in Alabio ducks, it decreased until the last observation day. Apoptosis percentage increased earlier in the Alabio duck than in domestic chicken.

Morphology, Biochemical, and Molecular Characterization of Pasteurella multocida Causing Hemorrhagic Septicemia in Indonesia.

Pasteurella multocida is a Gram-negative bacterium that causes hemorrhagic septicemia (HS) in buffaloes and cattle. The disease causes serious problems in Indonesian livestock and is classified as a serious transmissible animal disease. Previous research has determined the diversity of P. multocida using a serotyping method based on the antigenic properties of capsule polysaccharides. An alternative method for analysis utilizes sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and random amplified polymorphic DNA (RAPD). This study aimed to characterize and determine P. multocida diversity in several regions of Indonesia based on phenotypic character, protein profile, and the band pattern of RAPD results. Bacterial identification was performed using traditional biochemical techniques and API® 20NE systems and then confirmed molecularly using polymerase chain reaction (PCR). The freeze-thawing technique was performed to obtain the bacterial protein extract, and DNA extraction was executed using DNAzol. The extracted protein and RAPD product were then electrophoresed on 12% polyacrylamide gel and 1.5% agarose gel, respectively. The results indicate that the molecular weight range of the protein bands is 12-209 kDa, and the band pattern of the RAPD results ranged from 307-3,100 bp. Based on phenotypical analysis, P. multocida from South Sulawesi Province exhibited a variety of growth characteristics in MacConkey agar media. Using the hierarchical clustering analysis of the band patterns of RAPD and the whole-cell protein profiles, four and five clusters were formed, respectively. These results indicate molecular diversity among P. multocida from several regions of Indonesia.

Isoflavone and Antioxidant of Instant Cream Soup Made from Pumpkin and Tempeh and Their Active Compound in Ovariohysterectomy Rat-Induced Alzheimer's Disease.

Menopause period associated with brain function disorders can caused by decreasing estradiol levels and increasing oxidative stress in the body. Antioxidant agents are required to balance oxidative stress in the body. Instant cream soup made from pumpkin and tempeh is a supplementary food containing isoflavone and antioxidant agents. This study is aimed at analyzing the content of antioxidant level and isoflavones (genistein and daidzein) in instant cream soup and their effect on ovariohysterectomy (OVx) rats. Instant pumpkin cream soup with and without tempeh were subjected to isoflavone content and antioxidant analysis. Serum estradiol was analyzed using enzyme-linked immunosorbent assay. Liver concentration of malondialdehyde (MDA) and activities of superoxide dismutase (SOD) were measured by spectrophotometric methods. The study showed that the isoflavone content (genistein 370.86 g/100 g, daidzein 185.61 g/100 g) was only present in the pumpkin instant cream soup with tempeh (IPTS). IPTS has higher antioxidant levels (134.25 mg AEAC/100 g) than instant pumpkin cream soup without tempeh (IPS). In vivo study, experimental rats showed that OVx increased malondialdehyde (MDA) levels up to 5.85-6.07 nmol mL-1 as compared to control (4.47 nmol mL-1). Moreover, instant pumpkin cream soup with tempeh treatments significantly increased serum estradiol levels (2.37-3.63 µg) and superoxide dismutase (SOD) levels of 497.49-558.89 U mL-1. This study concluded that instant pumpkin cream soup and tempeh contained isoflavone and antioxidant, and it increased estradiol serum and SOD level.

Immunoglobulin yolk targeting spike 1, receptor binding domain of spike glycoprotein and nucleocapsid of SARS-CoV-2 blocking RBD-ACE2 binding interaction.

Coronavirus disease (COVID)-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has become a global pandemic disease that has social and economic chaos. An alternative mitigation strategy may involve the use of specific immunoglobulin (Ig)-Y derived from chicken eggs. Our study aimed to evaluate the neutralizing potential of specific IgY targeting S1, receptor-binding-domain (RBD) of spike glycoprotein and nucleocapsid (N) of SARS-CoV-2 to inhibit RBD and angiotensin-converting-enzyme-2 (ACE2) binding interaction. Hy-Line Brown laying hens were immunized with recombinant S1, RBD spike glycoprotein, and nucleocapsid (N) of SARS-CoV-2. The presence of specific S1,RBD,N-IgY in serum and egg yolk was verified by indirect enzyme-linked immunosorbent assay (ELISA). Specific S1,RBD,N-IgY was purified and characterized from egg yolk using sodium-dodecyl-sulfate-polyacrylamide-gel-electrophoresis (SDS-PAGE), and was subsequently evaluated for inhibition of the RBD-ACE2 binding interaction in vitro. Specific IgY was present in serum at 1 week post-initial immunization (p.i.i), whereas its present in egg yolk was confirmed at 4 weeks p.i.i. Specific S1,RBD,N-IgY in serum was able to inhibit RBD-ACE2 binding interaction between 4 and 15 weeks p.i.i. The results of the SDS-PAGE revealed the presence of bands with molecular weights of 180 kDa, indicating the presence of whole IgY. Our results demonstrated that S1,RBD,N-IgY was able to inhibit RBD-ACE2 binding interaction in vitro, suggesting its potential use in blocking virus entry. Our study also demonstrated proof-of-concept that laying hens were able to produce this specific IgY, which could block the viral binding and large production of this specific IgY is feasible.

Isolation and characterization of an orthoreovirus from Indonesian fruit bats.

Nelson Bay orthoreovirus (NBV) is an emerging bat-borne virus and causes respiratory tract infections in humans sporadically. Over the last two decades, several strains genetically related to NBV were isolated from humans and various bat species, predominantly in Southeast Asia (SEA), suggesting a high prevalence of the NBV species in this region. In this study, an orthoreovirus (ORV) belonging to the NBV species was isolated from Indonesian fruit bats' feces, tentatively named Paguyaman orthoreovirus (PgORV). Serological studies revealed that 81.2% (108/133) of Indonesian fruit bats sera had neutralizing antibodies against PgORV. Whole-genome sequencing and phylogenetic analysis of PgORV suggested the occurrence of past reassortments with other NBV strains isolated in SEA, indicating the dispersal and circulation of NBV species among bats in this region. Intranasal PgORV inoculation of laboratory mice caused severe pneumonia. Our study characterized PgORV's unique genetic background and highlighted the potential risk of PgORV-related diseases in Indonesia.

Atherosclerotic Lesion of the Carotid Artery in Indonesian Cynomolgus Monkeys Receiving a Locally Sourced Atherogenic Diet.

The atherosclerotic lesion is a principal hallmark of atherosclerotic animal models. This study aimed to assess lesions of the carotid artery in Indonesian cynomolgus monkeys exposed to an IPB-1 atherogenic diet. A total of 20 adult male cynomolgus monkeys received the local IPB-1 diet for two years. Blood lipid profiles, morphology, and carotid ultrasound of monkeys were measured. Nine of them were euthanized to confirm atherosclerotic lesions. Common carotid arteries (CCA) and carotid bifurcation (BIF) samples were collected and stained using Verhoef-van Giessen and CD68 immunohistochemistry. The results reveal the presence of severe atherosclerosis plaques in six out of nine animals (66.7%) corresponding to intermediately and hyper-responsive groups. The hyper-responsive group displayed the highest response in the developing intimal area (IA) at the CCA (0.821 mm2), whereas the hyporesponsive group had the smallest IA (0.045 mm2) (p = 0.0001). At the BIF, the hyporesponsive group showed the smallest IA (p = 0.001), but there was no difference between the intermediately and hyper-responsive groups (p = 0.312). The macrophage marker CD68 was also expressed on the cartotid of the intermediately and hyper-responsive groups. These results indicate that severe atherosclerotic lesions with high infiltration of macrophages were formed in the carotid arteries of intermediately and hyper-responsive Indonesian cynomolgus monkeys fed with the local atherogenic diet IPB-1 over two years, thus confirming atherosclerosis in a nonhuman primate model.

Effects of ethanol extract of curry leaves (Murraya koenigii) on HER2 and caspase-3 expression in rat model mammary carcinoma.

BACKGROUND AND AIM: Human epidermal growth factor receptor 2 (HER2/erbB2/neu) is a prognostic factor and biomarker for detecting mammary tumor malignancy. Leaves of curry (Murraya koenigii) contain alkaloid, flavonoid, and phenolic compounds that can be cytotoxic to tumor cells. Caspase-3 is an indicator of apoptosis in tumor cells. This study aimed to evaluate the effect of curry leaf extract on the expression of HER2 and caspase-3 in mammary tumor through immunohistochemical analyses. MATERIALS AND METHODS: Thirty five Sprague-Dawley rats were divided into seven groups: negative control of tumor (P1), positive control of tumor (P2), tumor therapy with methotrexate (P3), and curry leaf extract doses of 300 and 400 mg/kg body weight/BW after tumor formation (P4, P5), and before tumor formation (P6, P7). Thirty rats of six groups were injected subcutaneously into the mammary glands with 7,12-dimethylbenz(α)-anthracene DMBA) twice within 2 weeks for mammary tumor formation. At the end of the treatments, the rats were euthanized, and their mammary glands were analyzed histopathologically and immunohistochemically using HER2 and caspase-3 antibodies. RESULTS: Regarding the expression of HER2 detected in the epithelial cell membrane of the mammary gland, P2, P3, P4, and P5 revealed positive expression, P6 and P7 showed equivocal expression, while P1 showed negative expression. Regarding caspase-3 expression in the cytoplasm of epithelial cells, it was low in P1, moderate in P2, P5, P6, and P7, and high in P3 and P4. These findings suggest that DMBA injection produced mammary tumors with HER2 as a biomarker of mammary tumor, and high caspase-3 expression in P4 was the effect of curry leaves extract. CONCLUSION: The extract of curry leaves at a dose of 300 mg/kg BW with preventive and curative effects can potentially be used as an anti-tumor agent, which effectively induces the apoptosis of tumor cells.

First evidence of Klebsiella pneumoniae infection in Aceh cattle: Pathomorphology and antigenic distribution in the lungs.

BACKGROUND AND AIM: Klebsiella pneumoniae is an emerging zoonotic and foodborne pathogen worldwide. Hypervirulent K. pneumoniae (hvKp) was reported as the causative agent of bovine mastitis. This is the first study in Indonesia that has been conducted to determine the capsular serotype of K. pneumoniae, pulmonary gross pathology and histopathology, and distribution of hvKp in the lungs of Aceh cattle. MATERIALS AND METHODS: The presence of K. pneumoniae in Aceh cattle was investigated in two slaughterhouses in Banda Aceh and Aceh Besar, Indonesia. Lung tissues with gross pathological lesions were collected from 15 cattle presenting with depression, dehydration, or cachexia. The confirmation and capsular serotyping of K. pneumoniae isolates were performed using polymerase chain reaction. The tissues were stained with hematoxylin-eosin and immunohistochemistry to observe the histopathological lesions and the distribution of the hvKp antigens. RESULTS: The pneumonic lesions identified in the lungs of Aceh cattle included hyperemia, hemorrhage, consolidation, and atelectasis. K. pneumoniae was isolated in all 15 lung tissues with pathological pneumonic lesions. Two patterns of infection were observed histopathologically. Acute infection was characterized by hyperemia, inflammatory cell infiltration, hemorrhage, bronchiolar epithelium hyperplasia, bronchial and bronchiolar obstruction with purulent exudates, edema, and atelectasis. On the other hand, chronic infection was defined by macrophage infiltration, emphysema, bronchial dilatation, pleural fibrosis, and alveolar wall thickening by interstitial fibrosis. Immunohistochemical staining using monospecific antisera induced by the hvKp isolate confirmed the presence of K. pneumoniae-specific antigens in the acute infection, predominantly in the bronchiolar, vascular, and alveolar areas. In contrast, generally diffuse infiltrates were found in the pleura and interstitial alveolar areas in chronic infection. CONCLUSION: hvKp can be detected in the lungs of Aceh cattle, representing acute and chronic infections. The distribution of Klebsiella antigens in the lung tissue was consistent with the histopathological findings.

Characterization of a Novel Alphaherpesvirus Isolated from the Fruit Bat Pteropus lylei in Vietnam.

Herpesviruses exist in nature within each host animal. Ten herpesviruses have been isolated from bats and their biological properties reported. A novel bat alphaherpesvirus, which we propose to name "Pteropus lylei-associated alphaherpesvirus (PLAHV)," was isolated from urine of the fruit bat Pteropus lylei in Vietnam and characterized. The entire genome sequence was determined to be 144,008 bp in length and predicted to include 72 genes. PLAHV was assigned to genus Simplexvirus with other bat alphaherpesviruses isolated from pteropodid bats in Southeast Asia and Africa. The replication capacity of PLAHV in several cells was evaluated in comparison with that of herpes simplex virus 1 (HSV-1). PLAHV replicated better in the bat-originated cell line and less in human embryonic lung fibroblasts than HSV-1 did. PLAHV was serologically related to another bat alphaherpesvirus, Pteropodid alphaherpesvirus 1 (PtAHV1), isolated from a Pteropus hypomelanus-related bat captured in Indonesia, but not with HSV-1. PLAHV caused lethal infection in mice. PLAHV was as susceptible to acyclovir as HSV-1 was. Characterization of this new member of bat alphaherpesviruses, PLAHV, expands the knowledge on bat-associated alphaherpesvirology.IMPORTANCE A novel bat alphaherpesvirus, Pteropus lylei-associated alphaherpesvirus (PLAHV), was isolated from urine of the fruit bat Pteropus lylei in Vietnam. The whole-genome sequence was determined and was predicted to include 72 open reading frames in the 144,008-bp genome. PLAHV is circulating in a species of fruit bats, Pteropus lylei, in Asia. This study expands the knowledge on bat-associated alphaherpesvirology.

The contribution of domestic animals to the transmission of schistosomiasis japonica in the Lindu Subdistrict of the Central Sulawesi Province, Indonesia.

BACKGROUND AND AIM: Schistosomiasis is endemic in Indonesia and is found in three remote areas in Central Sulawesi Province. Non-human mammals serve as reservoir hosts, meaning the disease is zoonotic. The previous schistosomiasis studies in animals from the Lindu Subdistrict did not determine which domestic animal species can serve as the primary source of transmission. No animals have been treated in Indonesia to control the disease; therefore, the parasite's life cycle is not blocked entirely. This study aimed to determine the prevalence and identify the risk factors associated with, Schistosoma japonicum infection in animals, and identify animals' relative contributions to S. japonicum transmission in the Lindu Subdistrict. MATERIALS AND METHODS: A cross-sectional survey of S. japonicum infected animals was conducted in five villages of the Lindu Subdistrict. Fecal samples were collected from 134 selected animals (13 cattle, 26 buffaloes, 28 horses, 59 pigs, and 8 dogs). S. japonicum infection and infection intensity were determined using the Danish Bilharziasis Laboratory method. Environmental contamination with schistosome eggs was measured. The data were analyzed using a Chi-square test. RESULTS: The overall prevalence of schistosomiasis was 32.9%, with the prevalence of infection in each species of animal at 61.5% in cattle, 42.3% in buffaloes, 25.0% in horses, 35.6% in pigs, and 12.5% in dogs. Free-range pigs were 8.667 times more likely to have S. japonicum infection than pigs kept in cages. Buffaloes, cattle, and horses were the primary sources of S. japonicum egg contamination, with relative transmission indices of 59.15%, 22.80%, and 10.61%, respectively. CONCLUSION: Bovines and horses are the main contributors to schistosomiasis transmission in the Lindu Subdistrict. In conjunction with other schistosomiasis control programs, the government should treat infected animals living within endemic areas where there are high infection rates of S. japonicum.

The effect of propolis administration on fetal development.

Propolis is one of the bee products that widely used in health therapy. However, there has no study evaluating the developmental toxicity of propolis. This study was aimed to analyze the effect of propolis administration during pregnancy on fetal development. The pregnant mice were divided into five groups including control group (Tween 80 1%), low-dose (380 mg/kg b.wt.) and high-dose (1400 mg/kg b.wt.) of water extract of propolis from Banten (WEB), and low-dose (380 mg/kg b.wt.) and high-dose (1400 mg/kg b.wt.) of ethanol extract of propolis from South Sulawesi (EES). Propolis was administered for 18 days of gestation and then sacrificed to analyze the fetal development by examining external and skeletal abnormalities. The histopathological examination of placenta was also conducted. The result showed both low-dose groups did not inhibit fetal development. However, the high-dose of EES significantly reduced the weight, crown-rump of fetuses and increased the number of resorption (p < 0.05). Fetal weight was the only significantly reduced parameter of fetal growth in the highdose group of WEB (p < 0.05). The histopathological examination of placenta showed a reduction of labyrinth development in both high-dose groups. Dose of 380 mg/kg dose of Indonesian propolis is relatively safe for consumption during pregnancy.

Metabolic and morphometric changes in Indonesian cynomolgus monkeys (Macaca fascicularis) fed an atherogenic diet composed of locally sourced ingredients.

BACKGROUND AND AIM: This study was designed to determine the effects of a new atherogenic diet formulated at Institut Pertanian Bogor (IPB) (Bogor, Indonesia) on metabolic, morphometric, and carotid artery imaging of cynomolgus monkeys. MATERIALS AND METHODS: A total of 20 adult male cynomolgus monkeys fed IPB-1 atherogenic diet for 1 year. Total plasma cholesterol (TPC), high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides, and morphometric measurements were evaluated at baseline and monthly during the study. Carotid plaques and intima-media thickness (IMT) were measured using ultrasonography at baseline and after 8 months of treatment. RESULTS: This diet increased TPC, LDL, and TPC/HDL ratio and induced carotid atherosclerosis in this model. The TPC, LDL, and TPC/HDL ratio were positively associated; however, HDL was negatively associated with carotid plaques and IMT. CONCLUSION: The IPB-1 atherogenic diet formulated with locally and readily available ingredients provides an economically and scientifically feasible monkey model to study atherosclerosis in Indonesia and Southeast Asia.

Production and characterization of Newcastle disease antibody as a reagent to develop a rapid immunodiagnostic test tool.

AIM: This research was conducted to produce and characterize ND antibody as reagent candidate to develop a rapid immunodiagnostic test tool. MATERIALS AND METHODS: Four New Zealand White rabbits were used in this study and divided into two groups. First group was injected by Sato ND antigen, and second group was injected by genotype VII ND antigen. This study is divided into three steps: (a) ND antibody production, (b) ND antibody purification, and (c) ND antibody characterization. First group was rabbit injected by Sato NDV (5×108.25 egg lethal doses (ELD)50/ml) and second group was injected by genotype VII NDV (5×106.5 ELD50/ml). Antigen induction was performed by subcutaneous administrated for first (day 1) and second (day 14) injection and intravenous administrated for third (day 30) injection. Blood was collected on day 8 after third injection. RESULTS: Antibody production increased on second antigen injection and reached a peak on day 9 after second antigen injection. Sato and genotype VII ND antibody can be produced without adjuvant within 38 days with the highest titer 210. Based on antibody titer data, both antigens induced antibody production in a similar trend. The characterization antibody by SDS-PAGE indicated that molecular weight of immunoglobulin G (IgG) is 154.93 kDa (whole IgG), heavy chain 54.39 kDa, and light chain 27.74 kDa. ND antibodies have specificity to homologous and heterologous NDVs in varying virulence. CONCLUSION: Sato and genotype VII ND antibodies have been successfully produced within 38 days without adjuvant. Specificity of ND antibodies to NDVs in varying virulence and cross-reaction between Sato ND antibody and genotype VII ND antibody indicates that the characterized ND antibodies can be used as a reagent to develop rapid immunodiagnostic test tools.

Valproic Acid Exposure of Pregnant Rats During Organogenesis Disturbs Pancreas Development in Insulin Synthesis and Secretion of the Offspring.

Valproic acid (VPA) plays a role in histone modifications that eventually inhibit the activity of histone deacetylase (HDAC), and will affect the expressions of genes Pdx1, Nkx6.1, and Ngn3 during pancreatic organogenesis. This experiment was designed to study the effect of VPA exposure in pregnant rats on the activity of HDAC that controls the expression of genes regulating the development of beta cells in the pancreas to synthesize and secrete insulin. This study used 30 pregnant Sprague-Dawley rats, divided into 4 groups, as follows: (1) a control group of pregnant rats without VPA administration, (2) pregnant rats administered with 250 mg VPA on day 10 of pregnancy, (3) pregnant rats administered with 250 mg VPA on day 13 of pregnancy, and (4) pregnant rats administered with 250 mg VPA on day 16 of pregnancy. Eighty-four newborn rats born to control rats and rats administered with VPA on days 10, 13, and 16 of pregnancy were used to measure serum glucose, insulin, DNA, RNA, and ratio of RNA/DNA concentrations in the pancreas and to observe the microscopical condition of the pancreas at the ages of 4 to 32 weeks postpartum with 4-week intervals. The results showed that at the age of 32 weeks, the offspring of pregnant rats administered with 250 mg VPA on days 10, 13, and 16 of pregnancy had higher serum glucose concentrations and lower serum insulin concentrations, followed by decreased concentrations of RNA, and the ratio of RNA/DNA in the pancreas. Microscopical observations showed that the pancreas of the rats born to pregnant rats administered with VPA during pregnancy had low immunoreaction to insulin. The exposure of pregnant rats to VPA during pregnancy disturbs organogenesis of the pancreas of the embryos that eventually disturb the insulin production in the beta cells indicated by the decreased insulin secretion during postnatal life.

Detection of novel gammaherpesviruses from fruit bats in Indonesia.

Bats are an important natural reservoir of zoonotic viral pathogens. We previously isolated an alphaherpesvirus in fruit bats in Indonesia, and here establish the presence of viruses belonging to other taxa of the family Herpesviridae. We screened the same fruit bat population with pan-herpesvirus PCR and discovered 68 sequences of novel gammaherpesvirus, designated 'megabat gammaherpesvirus' (MgGHV). A phylogenetic analysis of approximately 3.4 kbp of continuous MgGHV sequences encompassing the glycoprotein B gene and DNA polymerase gene revealed that the MgGHV sequences are distinct from those of other reported gammaherpesviruses. Further analysis suggested the existence of co-infections of herpesviruses in Indonesian fruit bats. Our findings extend our understanding of the infectious cycles of herpesviruses in bats in Indonesia and the phylogenetic diversity of the gammaherpesviruses.

Pathology and immunohistochemistry study of Newcastle disease field case in chicken in Indonesia.

AIM: The aim of the study was to examine pathology and the distribution pattern of Newcastle disease virus (NDV) in internal organs of chickens from a field case using immunohistochemical staining. MATERIALS AND METHODS: 10 groups of broiler, layer, and domestic chicken were collected from necropsy room Division of Pathology, Bogor Agricultural University. These chickens were originated from West Java and collected based on pathologist diagnosis as suspect of Newcastle disease (ND). They were subsequently confirmed positive of ND with real-time-reverse transcription polymerase chain reaction assay. The respiratory, circulatory, digestive, lymphoreticular and central nervous systems were collected for histopathology examination. RESULTS: The gross pathology and histopathology changes were tracheitis, pneumonia, pericarditis, myocarditis, catarrhal proventriculitis, catarrhal enteritis, typhlitis, perihepatitis, pancreatitis, nephritis interstitial, splenitis, atrophy of Bursa Fabricius, and encephalitis. CONCLUSION: The distribution pattern of NDV in internal organs of chickens from a field case in this study is similar with a previous reported pattern in systemic cases of the internal chicken organs. High intensity of immunohistochemistry stain result was detected in trachea, lung, proventriculus, duodenum, cecal tonsil, kidney, and brain.

Changes in histopathology and cytokeratin AE1/AE3 expression in skin graft with different time on Indonesian local cats.

AIM: A good skin graft histopathology is followed by formation of hair follicle, sweat gland, sebaceous gland, blood vessel, lightly dense connective tissue, epidermis, and dermis layer. This research aimed to observe histopathology feature and cytokeratin AE1/AE3 expression on cat skin post skin grafting within a different period of time. MATERIALS AND METHODS: Nine male Indonesian local cats aged 1-2 years old weighing 3-4 kg were separated into three groups. First surgery created defect wound of 2 cm × 2 cm in size to whole groups. The wounds were left alone for several days, differing in interval between each group, respectively: Group I (for 2 days), Group II (for 4 days), and Group III (for 6 days). The second surgery was done to each group which harvested skin of thoracic area and applied it on recipient wound bed. On day 24th post skin graft was an examination of histopathology and cytokeratin AE1/AE3 immunohistochemistry. RESULTS: Group I donor skin's epidermis layer had not formed completely whereas epidermis of donor skin of Groups II and III had completely formed. In all group hair follicle, sweat gland, sebaceous gland, and neovascularization were found. The density of connective tissue in Group I was very solid than other groups. Cytokeratin AE1/AE3 expression was found on donor skin's epithelial cell in epidermis and dermis layer with very brown intensity for Group II, brown intensity for Group II, and lightly brown for Group I. CONCLUSION: Histopathological structure and cytokeratin AE1/AE3 expression post skin graft are better in Groups II and III compared to Group I.

Pathotypic characterization of Newcastle disease virus isolated from vaccinated chicken in West Java, Indonesia.

AIM: This research was conducted to differentiate and characterize eight Newcastle disease virus (NDV) isolates collected from vaccinated chicken at commercial flocks in West Java, Indonesia, in 2011, 2014 and 2015 by pathotype specific primers. MATERIALS AND METHODS: A total of eight NDV isolates collected from clinical outbreaks among commercial vaccinated flocks in West Java, Indonesia, in 2011, 2014, and 2015 were used in this study. Reverse transcription-polymerase chain reaction was used to detect and differentiate virulence of NDV strains, using three sets of primers targeting their M and F gene. First primers were universal primers to detect NDV targeting matrix (M) gene. Other two sets of primers were specific for the fusion (F) gene cleavage site sequence of virulent and avirulent NDV strains. RESULTS: Our results showed that three isolates belong to NDV virulent strains, and other five isolates belong to NDV avirulent strains. The nucleotide sequence of the F protein cleavage site showed 112K/R-R-Q/R-K-R/G-F117 on NDV virulent strains and 112G-K/R-Q-G-R-L117 on NDV avirulent strain. CONCLUSION: Result from the current study suggested that NDV virulent strain were circulating among vaccinated chickens in West Java, Indonesia; this might possess a risk of causing ND outbreaks and causing economic losses within the poultry industry.

Subjective and objective observation of skin graft recovery on Indonesian local cat with different periods of transplantation time.

AIM: The success of a skin graft in a cat is highly dependent on the granulation formed by the base of recipient bed. Granulation by the base of recipient bed will form after several days after injury. This research aimed to observe subjective and objective profile of skin graft recovery on forelimb of cats with different periods of donor skin placement. MATERIALS AND METHODS: Nine male Indonesian local cats aged 1-2 years old, weighing 3-4 kg were divided into three groups. The first surgery for creating defect wound of 2 cm×2 cm in size was performed in the whole group. The wound was left for several days with the respective interval for each group, respectively: Group I (for 2 days), Group II (for 4 days), and Group III (for 6 days). In the whole group, the second surgery was done by the harvesting skin of thoracic area which then applied on recipient bed of respective groups. RESULT: The donor skin on Group II was accepted faster compared to Group I and Group III. The donor skin did not show color differences compared to surrounding skin, painless, bright red in bleeding test had faster both hair growth and drug absorption. Test toward the size of donor skin and the effect of drugs did not show a significant difference between each group. CONCLUSION: The observe subjective and objective profile of skin graft recovery on forelimb of cats on Group II were accepted faster compared to Group I and III.