Effect of human umbilical cord stem cells (HUMSC) administration on collagen expression in the anterior vaginal wall in menopausal rats.

Objective: To evaluate the outcome of human umbilical cord stem cells (HUMSC) administration on collagen expression within the frontal vaginal wall of menopausal rats. Materials and Methods: We conducted an experimental, randomized post-test-only controlled group design. The study samples were 40 healthy female Winstar rat with the age of 8-12 weeks that had been ovariectomized, had never mated, and weighed 18-22 grams. The umbilical cord was obtained from voluntary donors who did not have a history of hepatitis B, hepatitis C, HIV, cytomegalovirus infection, treponema pallidum infection, or a history of other infections transmitted through the blood, placental tract, and genitals. Data collection (frontal vaginal wall of the rat) was carried out in a controlled environment with the consideration that all conditions were maintained equally and could be controlled. Results: There were 36 samples. A total of 13 menopausal rats (72%) had strong collagen expression and 5 rats had weak-to-moderate collagen expression (28%). On the other hand, 18 menopausal rats (100%) that belonged to the control group had weak-moderate collagen expression, and no menopausal rats appeared to have strong expression (0%). The administration of collagen to the anterior vaginal wall of postmenopausal rats proved to be effective by increasing the strong collagen expression in the damaged anterior vagina of postmenopausal female rats (p<0.05). Conclusion: Administration of HUMSC resulted in an increase in collagen levels in the anterior vaginal tissue of postmenopausal female rats. These results demonstrate significant therapeutic potential for the treatment of pelvic floor dysfunction.

Seroconversion, genotyping, and potential mosquito vector identification of Japanese encephalitis virus in pig sentinel settings in Bali, Indonesia.

Background and Aims: Despite the endemicity of Japanese encephalitis virus (JEV) in humans and animals in the Province of Bali, Indonesia, there is little data on whether seroconversion to the virus occurs in pigs, JEV genotypes circulating, and it's potential mosquito vectors in the area. The aims of this study were to (i) Determine whether JEV infection in Balinese pigs occurs before reaching their sexual maturity, (ii) identify the genotypes of circulating JEV, and (iii) identify potential JEV mosquito vectors at the study sites in urban and peri-urban areas of Bali. Materials and Methods: Sixteen 1-week-old Landrace piglets from two different sows were housed in Denpasar. Similarly, 18 one-week-old mixed-breed piglets of two different sows were housed in Badung Regency. The piglets were bled every 1 to 4 weeks for up to 24 weeks. Serum samples from the 11 piglets were tested for antibodies against JEV, and seroconversion-suspected sera were titrated using an enzyme-linked immunosorbent assay. Blood of seroconverted sera from pigs were tested using polymerase chain reaction (PCR) to detect the genetic sequence of JEV. The mosquitoes in the sentinels were trapped throughout the study period to identify the potential mosquito vectors of JEV. Results: Antibodies were detected in most of the selected piglets' sera from weeks 1 to 24 of their age. However, sera of pig B9 collected from the sentinel setting in Badung Regency showed a four-fold increase in antibody titer from week 4 to week 8, indicating seroconversion. PCR testing of blood from B9 (pooled blood sample collected from week 5 to week 8) identified JEV nucleic acids, which were phylogenetically classified as belonging to the JEV genotype III. Meanwhile, 1271 of two genera of mosquitoes, Anopheles spp. and Culex spp. were trapped in the pig sentinels. Conclusion: JEV seroconversion likely occurs before the pig reaches sexual maturity in Badung Regency. Sequence data indicate that JEV genotype III is circulating in the pig sentinel setting in the regency; however, circulating genotypes need to be clarified through increased surveillance. Meanwhile, Culex spp. and most likely Culex quinquefasciatus and Anopheles spp. were the dominant mosquitoes present in the study sites set in the urban area of Denpasar and peri-urban areas of Badung, Bali, indicating that these are likely vectors in spread of JEV in the region.

Purple sweet potato (Ipomoea batatas L.) extract effects on levels of inflammatory markers and chondrocyte count in gout arthritis Wistar rat model.

BACKGROUND: Recently, herbal medicine has become alternative in management of gout. Our aim is to assess effectiveness of purple sweet potato extract in gout. METHOD: In vivo study with randomized posttest only control group design. Purple sweet potato extract administered to 16 Wistar rats with MSU-induced gout. Independent t-test for analyzing interleukin-1 ß (IL-1ß), matrix metalloproteinase-3 (MMP-3), cartilage oligomeric matrix protein (COMP), malondialdehyde (MDA), and number of chondrocytes results. RESULTS: Decreased level of IL-1ß (3.81 ± 1.54 ng/mL vs. 2.55 ± 0.59 ng/mL, p = 0.04), MDA (5.04 ± 1.02 ng/mL vs. 2.27 ± 0.57 ng/mL, p = 0.04), MMP-3 (5.66 ± 1.02 ng/mL vs. 3.84 ± 1.37 ng/mL, p = 0.01) COMP (21.01 ± 3.57 ng/mL vs. 17.27 ± 2.60 ng/mL, p = 0.03), and increasing chondrocytes (35.17 ± 12.35 lp vs. 48.56 ± 7.17 lp, p = 0.02). CONCLUSION: Purple sweet potato extract with anthocyanin inhibits inflammation and cartilage degeneration in gout. LEVEL OF EVIDENCE: Level 1.

The Effects of Percutaneous Coronary Intervention on Biomarkers and Quality of Life in Patients With Chronic Total Coronary Artery Obstruction.

Background: Chronic total occlusion (CTO) is an angiographic picture of total occlusion without blood flow which is estimated to have lasted at least 3 months. This study attempted to provide an overview of the levels of matrix metalloproteinase-9 (MMP-9), soluble suppression tumorigenicity 2 (sST2), and N-terminal pro-B-type natriuretic peptide (NT-pro-BNP) as remodeling, inflammatory, and atherosclerotic markers, as well as changes in the angina severity in patients with CTO who underwent percutaneous coronary intervention (PCI) compared to those without PCI. Methods: This study is a preliminary report with quasi-experimental design study with a pre-test and post-test approach to compare PCI's effect in CTO patients towards changes in MMP-9, sST2, NT-pro-BNP levels, and changes in the angina severity. Twenty subjects underwent PCI and 20 subjects with optimal medical therapy, who were then assessed at baseline and 8 weeks after intervention. Results: The results of this preliminary report showed that decreased MMP-9 (pre-test: 12.07 ± 1.27 ng/mL vs. post-test: 9.91 ± 5.19 ng/mL, P = 0.049), sST2 (pre-test: 37.65 ± 20.00 ng/mL vs. post-test: 29.74 ± 15.17 ng/mL, P = 0.026) and NT-pro-BNP (pre-test: 0.63 ± 0.23 ng/mL vs. post-test: 0.24 ± 0.10 ng/mL, P < 0.001) levels were found after 8 weeks of PCI compared to those without such intervention. The levels of NT-pro-BNP were lower in the PCI group (0.24 ± 0.10 ng/mL) than in the non-PCI group (0.56 ± 0.23 ng/mL; P < 0.001). Moreover, there was an improvement of angina severity in PCI group than without PCI (P < 0.039). Conclusions: Although this preliminary report found a significant decrease in MMP-9, NT-pro-BNP, and sST2 levels in CTO patients who had undergone PCI compared to those without PCI, as well as improved angina severity in these patients, this study still has limitations. The number of samples was so small that similar studies with larger sample sizes or multicenter investigations are required to deliver more trustworthy and useful results. Nevertheless, we encourage this study as a preliminary baseline for further studies in the future.

Individual and Herd-Level Seroprevalence in Association with Potential Risk Factors of Japanese Encephalitis in Pigs Collected from Urban, Periurban, and Rural Areas of Bali, Indonesia.

A study to assess the seroprevalence antibodies against JEV in pigs in Denpasar, Badung, and Karangasem as the representatives of urban, periurban, and rural areas in the province of Bali was conducted. Sampled pigs' blood was collected and their sera were tested for antibody detection using commercial IgG ELISA. A standard questionnaire was used to interview the pig owners or farmers to identify the determinants associated with the seropositivity of the antibodies. Overall, 96.6% (95% CI: 94.5-98.1) of 443 pig sera in individual animal-level seroprevalence were seropositive to the ELISA. Karangasem had the highest test prevalence at 97.3% (95% CI: 93.1-99.2) while Badung had a slightly lower prevalence at 96.6% (95% CI: 92.2-98.9), and Denpasar had the lowest prevalence at 96% (95% CI: 91.5-98.5) (p=0.84). In herd-level seroprevalence, all sampled herds contained one or more seropositive pigs (overall herd-level seroprevalence 100% [95% CI: 97.7-100]). No animal-level factors were significantly associated with seropositivity (all p values >0.05). For the herd-level risk factors relating to pig management and husbandry practices adopted, no analysis model could be generated, as all the sampled herds were seropositive. More than 90% seroprevalence detected in this study indicates high natural JEV infection occurred in pigs, which highlights the high public health risk of the infection in the areas.

First survey on seroprevalence of Japanese encephalitis in long-tailed macaques (Macaca fascicularis) in Bali, Indonesia.

Background and Aim: Japanese encephalitis (JE) is a zoonotic infectious inflammatory brain disease caused by the JE virus (JEV). Considerable research into the seroprevalence of JE in domestic animals has been conducted, but there have been no reports of its occurrence in wild animals, including long-tailed macaques (Macaca fascicularis). This study aimed to estimate the seroprevalence of JEV infection and its determinants in long-tailed macaques in Bali and the prevalence of mosquito vectors. Materials and Methods: Blood samples (3 mL) were collected from a population of M. fascicularis (92 heads) inhabiting a small forest with irrigated rice field nearby (wetland area) in Ubud, Gianyar, and from two populations in dryland areas with no wet rice field (Uluwatu, Badung, and Nusa Penida, Bali Province, Indonesia). The collected sera were tested for antibodies against JEV using a commercially available enzyme-linked immunosorbent assay kit (qualitative monkey JE Immunoglobulin G antibody kit). The seropositivity of the antibodies was then compared based on different variables, namely, habitat type, age, and sex. Results: The seroprevalence of the JEV antibodies in all the samples tested was found to be 41.3%. The seropositivity of the monkey serum samples collected from the wetland area was 46.4%, which was higher than the seropositivity of the sera samples collected from the dried field areas (1.25%). Monkey sera collected from the wetland areas were 6.1 times (odds ratio [OR]: 6.1; 95% confidence interval [CI]: 0.71-51.5, p>0.05) more likely to be seropositive compared to the monkey sera collected from the dried field areas. Meanwhile, female monkeys were 1.79 times (OR: 1.79; 95% CI: 0.76-4.21; p>0.05) more likely to be seropositive to JEV than males. Similarly, juvenile monkeys were 2.38 times (OR: 2.38; 95% CI: 0.98-5.79); p>0.05) more likely to be seropositive against the JEV than adult monkeys. However, none of these differences achieved statistical significance. Regarding the JEV mosquito vector collection, more Culex mosquitoes were found in the samples from the wetland areas than from the dried field areas. Conclusion: The study confirms the existence of JEV infection in long-tailed macaques in Bali. There were patterned seropositivity differences based on habitat, age, and sex of the monkeys, but these were not significant. The possibility of monkeys as a JEV reservoir and the presence of the mosquitoes as the JEV vector are suggested but require more study to confirm.

The efficacy of binary ethylenimine-inactivated vaccines of Gianyar-1/AK/2014 virulent strain in protecting chickens against Tabanan-1/ARP/2017 virulent Newcastle disease virus isolates.

AIM: This study aimed to prepare binary ethylenimine (BEI)-inactivated virulent Newcastle disease virus (NDV) vaccine and to examine their ability to induce a protective antibody response in commercial chickens. MATERIALS AND METHODS: A virulent NDV field isolate Gianyar-1/AK/2014 was propagated in chicken-embryonated eggs and was then inactivated with BEI at a concentration of 4 mM. Three groups of chickens with low-level (2 log2 hemagglutination inhibition [HI] units) maternally derived antibodies against NDV were then immunized with the BEI-inactivated vaccine. A commercial live vaccine (LaSota strain) was used as positive control, and phosphate-buffered saline (PBS) was used as negative control. A challenge experiment with a virulent NDV of Tabanan-1/ARP/2017 was performed at 3 weeks post-vaccination. RESULTS: At 2 weeks post-immunization, the mean titers of antibodies against NDV in serum samples of chickens immunized with 0.2 mL of BEI-inactivated NDV (Group I), with live commercial NDV vaccine (Group II) and with PBS (Group III) were 3±0.94 log2 HI units, 4.9±0.99 log2 HI unit, and 0.0±0.0 HI units, respectively. At week 3 post-immunization, the mean titers of the antibodies for the three groups were 5±1.09 log2 HI units, 6.9±0.32 log2 HI units, and 0.00 HI units, respectively. The antibody titer induced by inactivated NDV Gianyar-1/AK/2014 isolates examined at 2 and 3 weeks post-vaccination was still at a significantly (p<0.01) lower level as compared to those induced by commercial life vaccine. However, the challenge test with virulent NDV of Tabanan 1/ARP/2017 isolates showed that all immunized chickens (Group I and II) survived without exhibiting any clinical sign post-challenge with the protection rates of 100%, whereas all chickens injected with PBS (Group III) died with clinical signs of ND. CONCLUSION: This finding shows that the BEI-inactivated vaccines prepared using virulent NDV of Gianyar-1/AK/2014 strain was able to induce protective antibody response in chickens but still at a lower level than those induce by commercial live NDV vaccine.

Antibody immunoglobulin G1 and immunoglobulin G2a responses against some cystic fluid proteins of Cysticercus bovis in Balb/c mice.

BACKGROUND AND AIM: Immunoglobulin (Ig) G1 and IgG2a are the surrogate markers respectively for humoral and cellular immune responses of hosts against antigens including cystic fluid proteins of Cysticercus bovis. A study was conducted to investigate the IgG1 and IgG2a responses of Balb/c mice against some individual cystic fluid proteins of C. bovis in an effort to determine the roles of each protein in inducing the humoral and cellular immune responses in host. MATERIALS AND METHODS: Individual p71, p31, and p14 proteins of C. bovis were purified by separation of the proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and elution of individual proteins from the gel. Six female Balb/c mice were immunized 4 times at 10-day intervals with the crude cystic fluid proteins, and sera were collected for the measurement of IgG1 and IgG2a levels against the individual proteins. Sera samples collected before the first immunization were used as negative antibody control, sera samples collected after the fourth immunization were used as positive antibody control, and crude cystic fluid protein was used as positive antigen control. RESULTS: All immunized mice were immune to p71, p31, p14, and crude cystic fluid proteins of C. bovis. The crude cystic fluid proteins of C. bovis induced a higher IgG2a than IgG1 level following the first and the second immunizations but switched into a higher IgG1 than IgG2a level following the fourth immunization. Protein 71 kDa (p71) induced a higher IgG2a than IgG1 level following the fourth immunization. In contrast, p14 induced a higher IgG1 than IgG2a level following the fourth immunization. Low and balance IgG1 and IgG2a levels against p31 were observed following the first to the fourth immunizations. CONCLUSION: Using IgG1 and IgG2a levels as the surrogate markers, it appears that cystic fluid antigens of C. bovis induce both humoral and cellular immune responses in Balb/c mice. The p71 appears to be a better inducer of cellular immune response, whereas p14 is a better inducer of humoral immune response of mice.

Antioxidant properties of topical Caulerpa sp. extract on UVB-induced photoaging in mice.

Caulerpa sp., a genus of seaweed native to the Indo-Pacific region, has been known for its antioxidant properties and health benefits when consumed as food. Previous studies have reported Caulerpa sp.'s potential as a strong antioxidant, but its effects on the skin in a topical preparation, especially its role in ultraviolet (UV) protection, have not been studied extensively. Our study investigated the protective effects of 0.2% and 0.4% Caulerpa sp. extract gels on photoaging in the UVB-irradiated skin of Wistar mice. The subjects were divided into naive control, vehicle control, and 3 treatment groups (0.2% Caulerpa sp. extract gel, 0.4% Caulerpa sp. extract gel, and 0.02% astaxanthin gel as a standard antioxidant). The groups, except the naive control group, received a total of 840 mJ/cm2 of UVB irradiation in four weeks. Protective effects of the extract were measured through the evaluation of collagen expression, matrix metalloproteinase (MMP)-1 expression and levels, and 8-OhDG expression. Mice who received topical application of Caulerpa sp. extract gel had higher collagen expression, better-preserved collagen structure, lower levels of MMP-1, and less MMP-1 and 8-OHdG expressions compared to the vehicle control group. There was no difference between different concentrations of the extract. Our findings demonstrated that topical application of Caulerpa sp. extract gel significantly protected UVB-irradiated mice skin from photoaging.

Protective antibody response of Balb/c mice to Bali rabies virus isolate propagated in BHK-21 cells.

The protective antibody response of Balb/c mice to Bali rabies virus (RABV) in BHK-21 cells was studied. The virus was isolated from a rabid dog and was adapted to replicate in BHK-21 cell culture for seven passages. The BHK-21-adapted Bali RABV (BHK-Bali RABV) was inactivated with binary ethylenimine and 24 mice were immunized twice at 21-days intervals with the inactivated virus and Rabisin® vaccine. Virus replication was detected using indirect immunofluorescence, immunocytochemistry, and western blotting assays. Enzyme-linked immunosorbent assay examination 2 weeks after the first immunization revealed RABV antibody titers that were mostly below the minimum protective level (<0.5 equivalent unit, EU). Antibody titers increased sharply after the second immunization. Antibody titers in serum of mice induced by inactivated BHK-Bali RABV one week after the second immunization were slightly lower (0.8-3.8 EU) than those induced by Rabisin vaccine (0.9-6.3 EU). RABV antibody titers were stable for at least 6 weeks after the second immunization. Both Rabisin vaccine and inactivated BHK-Bali RABV induced neutralizing antibodies with neutralization titers (50% protective dose per ml) of 29.84 for 0.1 ml Rabisin, 211.41 for 0.2 ml Rabisin, 27.41 for 0.1 ml BHK-Bali RABV, and 28.25 for 0.2 ml BHK-Bali RABV. Thus, inactivated BHK-Bali RABV induces a protective immune response in Balb/c mice, but at lower levels compared to induction by Rabisin vaccine.