Immunogenicity Response of Mus musculus var. Balb/C after Immunization using Flagellin Salmonella typhi Serovar Semarang.

<b>Background and Objective:</b> The flagellin of <i>Salmonella typhi</i> is potentially developed as an identifying antigen in a rapid diagnostic test instrument that may be more accurate than conventional serological tests. Therefore, this study aims to analyze the immunogenicity of flagellin <i>S. typhi</i> as the basis for developing a typhoid fever diagnostic. <b>Materials and Methods:</b> Flagellin was isolated from the bacterial culture of <i>S. typhi</i> serovar Semarang and used as the primary antigen for vaccine assembly. Native flagellin antigen was immunized in Balb/C mice with injection doses of 2, 3, 4, 5 and 6 g/100 L in each group (K0-K5), respectively, via intraperitoneal cavity. Blood serum was collected to ELISA based-measurement for IL-6 and TNF-a titers. Then, specific immunoglobulin (Ig) of anti-flagellin was detected using in-house ELISA and western blotting. <b>Results:</b> The findings in this study showed that immunization at the dose of 4-5 g/100 L significantly decreased the IL-6 titer, i.e., 8.33±0.87 pg mL¹, compared to control. The antibody titer test analysis showed the highest Ig-G anti-flagellin was found in K4 mice after immunization using a dose of 5 g/100 L with an average absorbance of Ig-G reaching 1.19 ±0.32. <b>Conclusion:</b> The results indicated that the flagellin protein of <i>S. typhi</i> serovar Semarang induces adaptive immune responses and produces specific antibodies against flagellin. The immunogenic properties of the flagellin protein of <i>S. typhi</i> serovar Semarang potentially developed as a specific diagnostic marker. Further research may also focus on a beneficial feature of flagellin as a vaccine candidate.

The Association of Tumor-Infiltrating Lymphocytes and Programmed Cell Death 1 Expression with the Incidence of Distant Metastasis in Triple-Negative Breast Cancer Subjects in Sanglah General Hospital, Bali, Indonesia.

The triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype with a high rate of distant metastasis. The tumor immunity microenvironment plays an important role, including tumor-infiltrating lymphocytes (TIL) and PD-1 (programmed cell death 1)/PD-L1 (programmed cell death-ligand 1), in promoting TNBC aggressiveness. This study aimed to determine the association of TIL and PD-L1 expression with the incidence of distant metastasis in TNBC. This study is a cross-sectional study involving TNBC subjects at Sanglah General Hospital, Denpasar, conducted in 2019. The parameters analyzed were the expression of TIL, PD-L1, and the incidence of distant metastasis. The expression of TIL was analyzed histopathologically while PD-L1 was measured with Ventana PD-L1 kit test. Subject characteristics were obtained from medical records. Data were collected and analyzed by SPSS 22.0. As many as 31 subjects with TNBC were included in this study, with 51.6% subjects with distant metastasis. The majority of subjects with distant metastasis had low TIL and low tumoral PD-L1 but high PD-L1 stromal in TIL. From statistical analysis, only PD-L1 stromal in TIL expression was associated significantly with distant metastasis (p = 0.043). In conclusion, there was a significant association between PD-L1 stromal in TIL and the incidence of distant metastasis in TNBC.

Preservation of semen from Kintamani Bali dogs by freezing method.

OBJECTIVE: To explore the effect of glycerol at different concentrations using different extenders on DNA fragmentation and motility of frozen-thawed Kintamani Bali dog spermatozoa. MATERIALS AND METHODS: Sample was collected from four mature Kintamani Bali dogs. Each ejaculate was prepared for cryopreservation with two different semen extenders; egg yolk Tris extender and coconut water-based extender. For each extender, three different glycerol concentrations were used; 4%, 6%, and 8%. Each of the six aliquots was loaded into 0.5 ml cryotube, placed on a styrofoam box 5 cm over liquid nitrogen for 10 min, and immersed in liquid nitrogen up to 8 min. Then, the frozen cryotubes were transferred into liquid nitrogen container. The cryotubes were thawed in a water bath at 38.5°C for 120 sec. After equilibration and thawing, each sample was assessed for motility parameters and for DNA fragmentation. RESULTS: The addition of 6% glycerol to extenders revealed the most effective addition of glycerol on motility and sperm DNA fragmentation after equilibrium and post-thawing. CONCLUSION: It is concluded that both extenders with the addition of 6% glycerol are safe to be used as an extender in Kintamani Bali dog semen preservation, and DNA fragmentation of Kintamani Bali dog spermatozoa was not influenced by the freezing procedure.

Group Counseling of Values Clarification to Increase Middle School Students’ Empathy / Terapia de grupo de clarificación de valores para incrementar la empatía de los estudiantes

This research compared the effectiveness of two techniques for administering group counseling focused on values clarification: modeling vs. role play. Effectiveness was measured in terms of participants' empathy at three time points, using a mixed factorial design. Participants were 40 students from a middle school in Mataram, Indonesia, who completed the Questionnaire of Cognitive and Affective Empathy (QCAE). Results of three ways mixed ANOVA showed that the two techniques of administering group counseling focused on values clarification were both effective in increasing participants’ empathy, although modeling appeared to be the most effective approach and is likely to be more efficient. Female students’ empathy was higher than males’, but there was no evidence that one group counseling technique worked better for girls than for boys. The results are discussed in terms of their implications for future studies and intervention

Esta investigación comparó la efectividad de dos técnicas para administrar terapia de grupo basada en la clarificación de valores: técnica de modelado frente a juego de roles. La efectividad se midió mediante la empatía de los participantes en tres momentos, empleando un diseño factorial mixto. Los participantes fueron 40 estudiantes de una escuela intermedia de Mataram, Indonesia, que cumplimentaron el Cuestionario de Empatía Cognitiva y Afectiva (QCAE). Los resultados de tres formas mixtas de ANOVA mostraron que las dos técnicas de administración de terapia de grupo centradas en la clarificación de valores fueron efectivas para aumentar la empatía de los participantes, aunque la técnica de modelado parecía ser el enfoque más eficaz y probablemente el más eficiente. La empatía de las estudiantes femeninas era más alta que la de los varones, pero no había evidencia de que una técnica de terapia de grupo funcionara mejor para las chicas que para los chicos. Se discuten los resultados en cuanto a su implicación para el estudio e intervención futuros

Timely initiation of breastfeeding is associated with the practice of exclusive breastfeeding in Indonesia.

BACKGROUND AND OBJECTIVES: Exclusive breastfeeding has been proven to be essential for optimal health, and for reducing infections and mortality in children. However, exclusive breastfeeding coverage both in Indonesia and in globalremains low. This study evaluated the relationship between the timely initiation of breastfeeding and the practice of exclusive breastfeeding in Indonesia. METHODS AND STUDY DESIGN: This cross-sectional study used Riskesdas 2013 data. Participants were 7,667 mothers whose children were aged 6-23 months in Indonesia, and were selected based on the completeness of the variables. The data were analysed using descriptive statistics, chisquare tests, and a multiple logistic regression that considered the sampling weight. STATA 13.0 was used for the analyses, and the significance level was set at p<0.05. RESULTS: Timely initiation of breastfeeding within 1 hour of parturition (OR=3.66, 95% CI: 2.14-3.64), timely initiation of breastfeeding at or after 1 hour following parturition (OR=2.79, 95% CI: 3.00-4.46), and neonatal illness (OR=0.69, 95% CI: 0.53-0.91) were significantly associated with an exclusive breastfeeding history among children aged 6-23 months. Other factors, such as the mother's age, mother's educational level, child's birth weight, household economic status, and residential area were not associated with an exclusive breastfeeding history. CONCLUSION: Timely initiation of breastfeeding and the prevention of neonatal illness should be the main interventions to improve exclusive breastfeeding coverage in Indonesia.

Antihyperglycemic activity of new 1,2,4-oxadiazolidine-3,5-diones.

A series of 1,2,4-oxadiazolidine-3,5-diones was synthesized and evaluated as oral antihyperglycemic agents in the obese insulin resistant db/db and ob/ob mouse - the two models for Type 2 diabetes mellitus. The majority of the prepared methoxy- and ethoxy-linked oxazole 1,2,4-oxadiazolidine-3,5-diones normalized plasma glucose levels at the 100 mg kg(-1) oral dose in the db/db diabetic mouse model, and several amongst them reduced the glucose levels at the 20 mg kg(-1) oral dose. The most potent compounds in the db/db mouse model were also active in the ob/ob mouse model normalizing the plasma glucose levels at the 20 mg kg(-1) oral dose. The trifluoromethoxy analog 32 was the most active compound of the series, reducing significantly the plasma glucose levels at the 5 mg kg(-1) oral dose. Oxadiazole-tailed 1,2,4-oxadiazolidine-3,5-diones were also active in both the db/db and ob/ob diabetic mouse models normalizing plasma glucose levels at the 100 mg kg(-1) oral dose.

2,4-Thiazolidinediones as potent and selective human beta3 agonists.

Methylsulfonamide substituted 2,4-thiazolidinedione 22c is a potent (EC50=0.01 microM, IA=1.19) and selective (more than 110-fold over beta1 and beta2 agonist activity) beta3 agonist. This compound has also been proven to be active and selective in an in vivo mode.

New azolidinediones as inhibitors of protein tyrosine phosphatase 1B with antihyperglycemic properties.

Insulin resistance in the liver and peripheral tissues together with a pancreatic cell defect are the common causes of type 2 diabetes. It is now appreciated that insulin resistance can result from a defect in the insulin receptor signaling system, at a site post binding of insulin to its receptor. Protein tyrosine phosphatases (PTPases) have been shown to be negative regulators of the insulin receptor. Inhibiton of PTPases may be an effective method in the treatment of type 2 diabetes. A series of azolidinediones has been prepared as protein tyrosine phosphatase 1B (PTP1B) inhibitors. Several compounds were potent inhibitors against the recombinant rat and human PTP1B enzymes with submicromolar IC(50) values. Elongated spacers between the azolidinedione moiety and the central aromatic portion of the molecule as well as hydrophobic groups at the vicinity of this aromatic region were very important to the inhibitory activity. Oxadiazolidinediones 87 and 88 and the corresponding acetic acid analogues 119 and 120 were the best h-PTP1B inhibitors with IC(50) values in the range of 0.12-0.3 microM. Several compounds normalized plasma glucose and insulin levels in the ob/ob and db/db diabetic mouse models.

Prodrugs of CL316243: a selective beta3-adrenergic receptor agonist for treating obesity and diabetes.

CL316243 is a highly selective and potent beta3-adrenergic receptor agonist, and has been shown in rodent models to be an effective agent for treating obesity and Type II diabetes. To improve the oral absorption and pharmacokinetic profiles of CL316243, a number of prodrugs have been synthesized and evaluated. Several ester-type prodrugs show significant improvements in oral bioavailability in both rodent and primate models.

Homogeneous Cell- and Bead-Based Assays for High Throughput Screening Using Fluorometric Microvolume Assay Technology.

High throughput drug screening has become a critical component of the drug discovery process. The screening of libraries containing hundreds of thousands of compounds has resulted in a requirement for assays and instrumentation that are amenable to nonradioactive formats and that can be miniaturized. Homogeneous assays that minimize upstream automation of the individual assays are also preferable. Fluorometric microvolume assay technology (FMAT) is a fluorescence-based platform for the development of nonradioactive cell- and bead-based assays for HTS. This technology is plate format-independent, and while it was designed specifically for homogeneous ligand binding and immunological assays, it is amenable to any assay utilizing a fluorescent cell or bead. The instrument fits on a standard laboratory bench and consists of a laser scanner that generates a 1 mm(2) digitized image of a 100-µmm deep section of the bottom of a microwell plate. The instrument is directly compatible with a Zymark Twistertrade mark (Zymark Corp., Hopkinton, MA) for robotic loading of the scanner and unattended operation in HTS mode. Fluorescent cells or beads at the bottom of the well are detected as localized areas of concentrated fluorescence using data processing. Unbound flurophore comprising the background signal is ignored, allowing for the development of a wide variety of homogeneous assays. The use of FMAT for peptide ligand binding assays, immunofluorescence, apoptosis and cytotoxicity, and bead-based immunocapture assays is described here, along with a general overview of the instrument and software.

Azole phenoxy hydroxyureas as selective and orally active inhibitors of 5-lipoxygenase.

Azole phenoxy hydroxyureas are a new class of 5-lipoxygenase (5-LO) inhibitors. Structure-activity relationship studies have demonstrated that electronegative substituents on the 2-phenyl portion of the oxazole tail increased the ex vivo potency of these inhibitors. Similar substitutions on the thiazole analogs had only minor contribution to the ex vivo activity. The trifluoromethyl-substituted oxazole 24 was the best compound of the oxazole series in both the ex vivo (6 h pretreated rats) and in vivo (3 h pretreated rats) RPAR assay with ED50 values of approximately 1 and 3.6 mg/kg, respectively, but was weakly active in the allergic guinea pig assay. Oxazole 50 was equally active in both the RPAR and guinea pig in vivo models and was similar to zileuton. The unsubstituted thiazole 52 was the best compound of the thiazole series, by inhibiting the leukotriene B4 biosynthesis in the RPAR assay (3 h pretreated rats) by 99%, at an oral dose of 10 mg/kg, and the bronchoconstriction in the allergic guinea pig by 50%, at an intravenous dose of 10 mg/kg. Oxazole 24 demonstrated high and selective 5-LO inhibitory activity in the in vitro assays, with IC50 values ranging from 0.08 microM in mouse macrophages to 0.8 microM in human peripheral monocytes to 1.2 microM in human whole blood. This activity was selective for 5-LO, as concentrations up to 15 microM in mouse macrophages did not affect prostaglandin formation. Oxazole 59 was the most active inhibitor in the human monocyte assay with an IC50 value of 7 nM.