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Objective: Validation of analysis method is important, especially in analyzing narrow-index drugs such as warfarin (WF). This study aimed to obtain a validated method of analyzing warfarin in human plasma according to European Medicine Agency guidelines. Methods: The optimum conditions for the analysis of warfarin in human plasma using fluorescence detector HPLC with Chiralcel OD-RH column (4.6 x 150 mm i.d., 5μm); Chiralcel OD-RH guard column (4.0 x 10 mm, 5μm), column temperature 45 °C. The mobile phase used was acetonitrile: phosphate buffer pH 2 (40:60), with an isocratic flow rate of 1 ml/min and an injection volume of 20 μl. Excitation and emission wavelengths were 310 and 350 nm (warfarin) and 300 and 400 nm (griseofulvin). The retention time of griseofulvin was 6-7.5 min; R-warfarin was 10-11.5 min; S-warfarin was 14-16 min. Results: The result of this validation obtained the optimum condition. This method yielded LOD values of 0.0674 ppm (R-warfarin) and 0.0897 ppm (S-warfarin). LOQ values were 0.225 ppm (R-warfarin) and 0.298 ppm (S-warfarin). Linearity at concentrations of 0.2-3 ppm with the line equation y = 0.0705x+0.0704 with R2 = 0.978 for R-warfarin and y = 0.0513x+0.0297 with R² = 0.9924 for S-warfarin. 75% of the seven concentrations met the reverse concentration requirements, which were below±15%. This method met the requirements of accuracy and precision within and between runs, selectivity and carryover where the %RSD and %diff values were below±15% Conclusion: This analytical method can be declared valid and can be used for sample measurement in warfarin pharmacokinetic studies.

Objective: Sonneratia alba leaves were used by the community for traditional medicine to cure muscle pain, back pain, antioxidants, rheumatism, malaria, wounds, tuberculosis (TB) and as a spermicide. S. alba leaves extract was easy to damage because of the light exposure, change of pH, weather and a long period of storage time. The problem can be solved by coating the extract with a microencapsulation technique. The purpose of this research was to formulate the microcapsules of S. alba leaves extract with solvent evaporation technique using Ethocel 10 cP and Eudragit E100 as a matrix. Methods: S. alba leaves were extracted using ethanol 96%. This extract was dried by a rotary evaporator. The microencapsulation process of S. alba leaves extract was done by solvent evaporation technique (O/W: oil in water). The formula of S. alba leaves extract microcapsules was designed into six formulas (Eudragit E100: EA1, EA2, EA3 and Ethocel 10 cP: EB1, EB2, EB3). Microcapsules of S. alba leaves extract were characterized for particle size in terms of surface morphology by scanning electron microscope (SEM) and encapsulation efficiency. Antioxidant activity of the formulation have been evaluated by DPPH method. Physical characterization on microparticles was performed by conducting entrapment efficiency and SEM picture. Results: In this research, the microparticles containing S. alba extract has been developed by using ethyl cellulose (Ethocel 10 cP) and eudragit (Eudragit E100) as the polymer matrix. The results showed that a high concentration of polymer (Ethocel 10 cP and Eudragit E100) used in microencapsulation resulted in better S. alba leaves extract microcapsules in terms of physical characteristics. Particle size of microcapsules containing S. alba leaves extract were in the range of 0.701 to 1.163 μm. Encapsulation efficiency (% EE) was categorized as poor because the value were ≤ 80% to which 74.386% (EB3) and 75.248% (EA1). SEM picture of EA1 (Eudragit E100) revealed that the surface of microcapsule were rough and porous. When Ethocel 10 cP was used as a polymer, a smoother surface and less visible pores of microcapsule were obtained. The antioxidant ability of S. alba leaves extract microcapsule showed that IC50 values were 53.26 ppm. Conclusion: It can be concluded that microcapsules of S. alba leaves extract can be prepared by solvent evaporation technique using Eudragit E100 and Ethocel 10 cP as polymer. S. alba leaves has potent antioxidant activity either as an extract or after being formulated into microcapsules.

Warfarin is a type of vitamin K antagonist anticoagulant used to thromboembolic prevention and its maintenance dose differs significantly among patients. Moreover, the factor influencing the dose calculation is known as genetic polymorphism, while the influence of vitamin K intake while the effect of vitamin K intake is still a matter of debate. This study aims to provide an overview of several studies on the effect of vitamin K intake on the stability of the PT-INR value and anticoagulation in warfarin therapy patients. A study was conducted on the effect of vitamin K intake on the PT-INR value stability and anticoagulation in warfarin therapy patients. One study showed that there was no significant correlation between vitamin K usual intake and the anticoagulant effect, while in another, it was found that the stability of this effect is achievable with adequate amounts of the vitamin intake. Furthermore, another study showed that the anticoagulant effect is more stable in patients with excessive vitamin K intake on long-term warfarin use. However, other study showed that INR values will stabilize if vitamin K intake is relatively low. The results from various studies were different, therefore, there is a debate regarding the influence of vitamin K intake on the PT-INR value and anticoagulation.

Objective: This study aimed to predict the effect of SNPs CYP2C9 s on the metabolic activity of S-warfarin in the body. Methods: Molecular modeling was performed to obtain SNPs CYP2C9 and molecular docking was performed to predict the effect of SNPs CYP2C9 on the metabolic activity of S-warfarin. Results: The results showed that wild-type CYP2C9 had the strongest binding affinity (∆G: -9.76 kcal/mol), indicating that wild-type CYP2C9 had the best metabolic activity compared to SNPs CYP2C9. There was a decrease in hydrogen bond formation in SNPs CYP2C9 and an increase in the distance between C7 S-warfarin and Fe-Heme in CYP2C9 SNPs when compared to wild-type CYP2C9 Conclusion: The decrease in binding affinity, decrease in hydrogen bond formation, and an increase in the distance between C7 S-warfarin and Fe-Heme on SNPs CYP2C9 indicated that SNPs CYP2C9 had decreased metabolic activity against S-warfarin which led to an increased risk of bleeding.

The increase in resistance of Escherichia coli bacteria that caused urinary tract infections to antibiotics narrows the selection of effective treatment which encourages the development of treatment therapies using natural ingredients. Maja fruit [Aegle marmelos (L.) Correa] is a plant that has shown antibactericidal activity against gram-negative bacteria, one of which is Escherichia coli. This study aimed to determine the antibacterial activity of fruit Maja extract, minimum inhibition concentration (MIC) and the minimum bactericidal concentration (MBC) of fruit Maja extract and the comparative value of gentamycin against Escherichia coli resistance isolated from urinary tract infection patients. The methods are extraction of Maja fruit by using 96% ethanol, agar diffusion method with paper disc and determination of MIC and MBC using microdilution method. The results showed that the ethanol extract of Maja fruit had antibacterial activity with value of MIC50 as 1400-1500 μg/ml, the value of MIC90 was 1600 μg/ml and the value of MBC was in the range concentration of 1700-2000 μg/ml against E. coli isolate I. Against E. coli isolates II, value of MIC50 was in concentration of 2900 μg/ml, MIC90 was in concentration 3000 μg/ml and the value of the MBC is in range 3150-3500 μg/ml. The comparative values of the antibacterial activity of gentamycin on fruit maja extract are 1:5.871 x 103.

Objective The risk of contracting tuberculosis (TB) and the efficacy of TB therapy are affected by several factors, including genetic variation among populations. In the Indonesian population, data on the genes involved in drug transport and metabolism of TB therapy are limited. The aim of this study was to identify the genetic profile of the ABCB1 gene (rs1128503 and rs1045642) and CYP2E1 gene (rs3813867) in Indonesians with TB. This study was a cross-sectional study of 50 TB outpatients in Jambi city, Indonesia. Sociodemographic characteristics were obtained from medical records. Whole blood was collected, and genomic DNA was isolated. Single nucleotide polymorphisms were determined using polymerase chain reaction-restriction fragment length polymorphism with HaeIII, MboI, and PstI for rs1128503, rs1045642 (ABCB1), and rs3813867 (CYP2E1), respectively. Result The frequency of alleles of each gene was analyzed by Hardy–Weinberg equilibrium. The genetic profiles of ABCB1 rs1128503 and rs1045642 were varied (CC, CT, TT), while CYP2E1 rs3813867 was present in CC (wild type). The genetic variations of ABCB1 and CYP2E1 may have no significant correlation with the duration of TB therapy. Nevertheless, this study may provide as preliminary results for the genetic profiles of ABCB1 (rs1128503, rs1045642) and CYP2E1 (rs3813867) in the Indonesia population.

Seborrhea dermatitis is a skin disorder that usually appears on parts of the body that have high density of sebaceous glands, such as the face, chest, and scalp. Clinical manifestations that generally appear as scaly skin and erythema. Seborrhea dermatitis is also known as one of the causes of alopecia. Treatments that can be used for seborrhea dermatitis are antifungal, anti-inflammatory, keratolytic, and coal tar. There are concerns about poor adherence, resistance, and some side effects of drugs that have been used in the treatment of seborrhea dermatitis. Concerns regarding these issues increase the urgency for the development of new therapeutic agents in the treatment of seborrhea dermatitis. Research on medicinal plants has enormous potential to produce compounds with new structures and bioactivity. This review discusses clinical and in vitro studies related to the activity of several medicinal plants that have potential as a treatment for seborrhea dermatitis, as well as the compounds that play a role in these activities. Literature searches were carried out on the PubMed, TaylorFrancis, and SpringerLink databases using Boolean Operators to get 25 articles that match the keywords used. Of the 25 articles, six were clinical trials, while 19 were in vitro studies of Malassezia. Several plants have potential as promising therapeutic agents for the treatment of seborrhea dermatitis by inhibiting the growth of Malassezia, decreasing sebum secretion, and decreasing symptoms associated with seborrhea dermatitis such as itching, pain or burning sensation, and redness.

Objectives: In patients with breast cancer, Human Epidermal Growth Factor is over expressed until 30%. Monoclonal antibodies was an alternative detection cancer in molecular level. The aim of the experiment was protein recombinant of anti-HER2 scFv was constructed from the gene encoding single chain variable fragment of anti-HER2 antibody wich was fused with Histag and can be expressed in the Eschericia coli BL21(DE3) to be used as a diagnostic protein for breast cancer cells. Methods: The recombinant pJ401express_anti-HER2 scFv fused with histaq was transformed into E. coli BL21 (DE3) and expressed as recombinant anti-HER2 scFv protein with various inducer concentration. Then, those protein was purified with the nickel polyhistidine tag (Ni-NTA) affinity chromatography using imidazole concentration i.e 100 and 150 mM. Finally, the existence of this recombinant protein was determined with anti histaq antibody in western blot assay. Results: Plasmid isolation from E. coli BL21 (DE3) cells revelaed the existence of the recombinant pJ401express_anti-HER2 scFv. The optimum condition for using IPTG as inducer for the intracellular expressed anti-HER2 scFv gene was 1 mM IPTG which was entered into broth medium at the 3.5th hr of growth time of E. coli BL21(DE3). Then, the higher amout of more purified anti-HER2 scFv was obtained using imidazole at 150 mM. The recombinant protein was also bound to anti histaq antibody in western blot assay. Conclusion: the recombinant pJ401express_anti-HER2 scFv was successfully expressed as anti-HER2 scFv protein.

Antibiotic resistance is a major global health problem and continues to increase, so there is a need forbetter evaluation and therapeutic advice for Urinary Tract Infection (UTI) patients. It had isolated E. colifrom clinical isolates that were resistant to antibiotics of β-lactam, the first generation of cephalosporinand fluoroquinolone. Characterization of bacterial sensitivity should be analyzed to other antibioticswith different mechanisms. Therefore, this study aims to analyze the antibiotic activity of fosfomycin,kanamycin, rifampicin, ceftriaxone, imipenem, and vancomycin against to resistant Escherichia coliclinical isolates of UTI. The sensitivity test in this study used the Kirby-Bauer disc diffusion methodand data analysis using SPSS version 24. The results revealed the diameter of inhibition zone (mm) ofantibiotics of fosfomycin, ceftriaxone, imipenem, kanamycin, rifampicin, and vancomycin respectivelyi.e. 38±1; 36.3±1.15; 35±1; 31.6±1,15; 28±1; and 26.3±0.17. Based on statistical analysis there weresignificant differences in the activity of fosfomycin, ceftriaxone, imipenem, kanamycin, rifampicin,and vancomycin.

In this research, Escherichia coli BL21 (DE3) harboring an expression vector constructed with a rhamnose-inducible promoter and a pelB signal peptide was used as a host cell to produce MPT64 protein. The objective of this research was to figure out the optimum time of mpt64 gene expression through real-time monitoring of MPT64 protein production and distribution in host compartments. The mpt64 expression was regulated by the rhamnose presence at a concentration of 4 mM. The real-time isolated protein was monitored using polyacrylamide gel electrophoresis in denaturation condition. Based on real-time monitoring, the MPT64 protein (24 kDa) in the cytoplasm was optimum detected at 24 h after induction. For periplasmic fraction, the protein was detected at 4 h after induction but thinning at 15 h after induction. At 16 h after induction, the MPT64 protein band was found in the medium with increasing concentrations until 24 h. Thus, it can be concluded that the mpt64 gene expression was regulated in the presence of rhamnose as an inducer, and the proteins were shown to be translocated throughout the host cell compartment with different levels of protein accumulation at different times, according to the role of pelB as a signal peptide.

Background Central obesity is a risk factor for metabolic syndrome. Subjects with central obesity have a higher risk of developing type 2 diabetes and cardiovascular disease. Many conditions affect the prevalence of central obesity, including energy expenditure, aging, proinflammatory conditions, and hormonal, genetic, and ethnic differences. Polymorphism of the APM1 gene, encoding the protein adiponectin, is closely related to metabolic syndrome. Adiponectin influences fatty acid oxidation and glucose intake in muscle. Therefore, variation in the APM1 gene is associated with diabetes and obesity. Purpose The aim of the present study was to investigate the correlation of the single-nucleotide polymorphism (SNP) of the APM1 SNP rs2241766 with body mass index (BMI) and lipid profiles in Indonesian (Bandung) subjects. Patients and methods Genotyping of the APM1 gene was performed using the Amplification Refractory Mutation System. Whole blood and serum of 54 subjects with central obesity (waist circumference [WC] ≥90 cm) and 53 healthy subjects (WC

Penderita kanker global diperkirakan telah mengalami peningkatan setiap tahunnya. Seiring dengan banyaknya kemajuan dalam pengobatan kanker, pengembangan menggunakan pendekatan baru untuk diagnosis kanker yang akurat dan untuk terapi yang ditargetkan berdasarkan penanda spesifik kanker. Nanomedicines bisa menjadi kandidat ideal untuk mencapai tujuan ini. Berbagai nanopartikel dikembangkan untuk pengiriman yang ditargetkan agen diagnostik / terapeutik ke situs kanker, dimaksudkan untuk menghasilkan efektivitas yang lebih besar dan efek samping yang lebih kecil. Biosensor perlu dikembangkan sebagai perangkat bioanalitik yang terkait dengan atau terintegrasi dengan transduser fisikokimia untuk analisis biomarker. Konsep penggunaan nanopartikel dalam pengembangan biosensor untuk diagnosis biomarker akan membuat perangkat ini semakin sensitif. Metode diagnostik yang sederhana dan sensitif yang dapat mendeteksi banyak biomarker kanker akan membantu dalam diagnosis dini dan meningkatkan tingkat kelangsungan hidup pasien kanker.

Breast cancer screening tests could reduce the mortality rates for breast cancer patients. Screening and detection are the keystone of cancer prevention and may significantly minimize the death rates in breast cancer patients for long-term. In this review, we would like to present a comprehensive summary from recent publications of the current development for breast cancer screening, classification of breast cancer based on pathological diagnosis, as well as development of breast cancer detection. The sources of the articles were collected from research published in the PubMed, NCBI databases and manual searches without time restriction based on review of the title, abstract and full review of the articles, using the keywords "breast cancer", "diagnostic", "screening", "imaging", "biomarker" and the combination of these terms. The criteria excluded in selecting references were articles that are not written in English, newspapers, and posters. Of the 146 articles that were selected, there were 103 articles included. Breast cancer screening consists of imaging and pathological assessment such as invasive biopsies of tumor tissue and measurement of biomarkers. The recent development of breast cancer screening utilizing different models and methods like biomarkers were being reviewed. For imaging methods, there are mammography, digital breast tomosynthesis (3D mammography), magnetic resonance imaging (MRI), and ultrasonography. For pathological assessment, there are primary biomarker analysis for breast cancer (estrogen receptor, progesterone receptor, HER2, KI67 index) and liquid biomarker analysis from blood or saliva samples. Additionally, there are some diagnostic kit models for breast cancer screening that were in use such as NanoString nCounter®, MammaTyper®, CellSearch System™, and AdnaTest BreastCancer™. Each of these methods has its own limitations. Therefore, the development of breast cancer models should be more sensitive, reliable, approachable and less harmful.

The use of warfarin was monitored using the International Normalized Ratio (INR) value. This study aims to determine the relationship between the patient’s demographic variables and INR, as well as the therapy result and differences in their INR value. A cross-sectional study was used with retrospective observational data collected by tracing secondary information of medical records from outpatients with heart disease on warfarin therapy at Dr. Hasan Sadikin hospital Bandung between 2016 to 2020. Subsequently, the difference in patients’ INR value was analyzed with the Mann-Whitney test based on dosage. At the same time, the relationship between demographic variables and INR was examined using the Chi-square test and Spearman correlation. The INR examination results in 192 subjects showed 124 patients (64.6%) reached the target with an average value of 2.37 ± 0.21 and a warfarin dose of 19.31 ± 6.25 mg weekly, while 68 (35.4%) had an average of 1.82 ± 0.73 and a dose of 20.24 ± 6.24 mg weekly. Subsequently, the difference test with Mann Whitney (p = 0.004) showed a variation in the INR value based on the warfarin dose. The Spearman correlation analysis results indicated a relationship between BMI (p = 0.009), daily (0.010), and weekly dose (0.008) on the INR value with the correlation coefficient of 0.188; 0.186; and 0.192; indicating a negative association with very weak correlation strength. This study showed that a majority of patients reached the INR value of 2-3, and the demographic variables associated with this ratio were BMI and warfarin dose. The decision to increase and decrease the warfarin dose is determined by INR value and BMI.

This study was aimed to isolate and characterize Pseudomonas aeruginosa antibiotic resistance profiles that isolated from bathroom water of five hospitals in Bandung, Indonesia, with different types of water reservoirs. Total of 25 water samples from bathrooms of five hospitals were collected and analyzed for the existence of P. aeruginosa colonies on the surface of MacConkey agar media using a streak plate method and identified using phenotypic identification and a series of biochemical tests. All P. aeruginosa isolates were tested against ceftazidime, piperacillin/tazobactam, ciprofloxacin, meropenem, and gentamicin containing in paper disc, using the agar diffusion method. Of all samples, the total number of P. aeruginosa isolates was less than that of non-P. aeruginosa. In hospitals that use permanent bathtubs, a greater total bacterial count was obtained than those using pails. From 110 isolates, 14.54% were multidrug resistance antibiotics. The majority of the resistant isolates were from hospital B with permanent bathtubs. Of 25 isolates from that hospital, P. aeruginosa isolates were resistant to ceftazidime (20%), piperacillin/tazobactam (4%), ciprofloxacin (20%), and gentamicin (20%). The multiple antibiotic resistance index value of P. aeruginosa isolates was 0.4–0.6. Thus, it can be concluded that the bathroom wáter in the hospital with permanent bathtubs were potential reservoirs of antibiotic-resistant P. aeruginosa.

Context: Human Epidermal Growth Factor (hEGF) is a potential therapeutic protein that has been widely used as a healing agent for various chronic wounds. It induces the proliferation and metabolism of epithelial cells, regenerates skin cells, and validates skin elasticity. In the previous study, recombinant hEGF (rhEGF) had been successfully expressed extracellularly in Escherichia coli (E. coli) BL21 (DE3) using pectate lyase B (PelB) signal peptide. The previous study has shown that the medium concentration and the induction time influenced the production of rhEGF. Aims: Therefore, this study was conducted to optimize the induction time and medium concentration for rhEGF extracellular secretion then followed by scale-up production. Settings and Design: This experiment was carried out using E. coli BL21 (DE3) which contains pD881 plasmid that carries hEGF and PelB gene. Optimization design of induction time and medium concentration were obtained using Central Composite Design (CCD). Methods and Material: The method of study started by the rejuvenation of E. coli culture, extracellular secretion, and optimization in the flask scale then followed by scaled-up production with high-cell density culture in the fermenter. Statistical analysis used: The optimization was carried out using Response Surface Methodology (RSM) and multi regression analysis. Results: This work showed that the multiplication of 1.5-fold medium concentration with induction time 3h after the culture started gave the best result among another condition in this study. Additionally, the rhEGF production in the fermenter scale was identified by SDS-PAGE Tricine and quantified by ELISA, which showed 122.40 μg of the rhEGF per milliliter medium. Conclusions: In respect of the result, we conclude that the optimized condition of extracellular secretion was successfully obtained, and gives higher result before the previous study.

In a previous study, 2′,4′-dihydroxy-6′-methoxy-3′,5′-dimethylchalcone (ChalcEA) isolated from the leaves of Eugenia aquea was reported to inhibit proliferation of the breast adenocarcinoma MCF7 cell line and to promote apoptosis via activation of poly(adenosine diphosphate-ribose) polymerase protein. The present study aimed to evaluate the inhibitory effect of ChalcEA on the proliferation of A549 lung cancer cells using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxylmethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, and to examine the ability of ChalcEA to induce apoptosis through activation of the caspase cascade signaling pathway in a western blotting assay. The results revealed that ChalcEA inhibited proliferation of the A549 lung cancer cell lines in a time- and dose-dependent manner with IC50 values of 25.36 and 19.60 µM for 24 and 48 h treatments, respectively. Western blot analysis indicated that ChalcEA exerted its anti-proliferative effects by promoting apoptosis via the activation of caspase-9 and caspase-3. Based on in silico results, ChalcEA with the binding energy of −6.53 kcal/mol could compete better than 4-methyl benzenesulfonamide (−6.43 kcal/mol) as an inhibitor of caspase-3 (PDB: 2XYG). ChalcEA has potential since it has three hydrophobic features. These results provided a basis for further study of ChalcEA as an active compound for anticancer therapeutics.

Methicillin-resistant Staphylococcus aureus (MRSA) is the most common bacteria causing nosocomial infections with high levels of resistance to available antibiotics. So, it is necessary to search for new compounds to solve this problem. Various studies showed antibacterial activity of rambutan peel but for Rambutan Binjai peel extract that are from Indonesia has never been studied against the MRSA. This study aims to determine the antibacterial activity, the value of minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) using agar diffusion method. The concentration of rambutan peel ethanol extract at as much as 62.5 mg/ml showed the inhibitory diameter i.e 21.3 ± 2.4 mm. MIC and MBC were in the same range, which was between 0.98 (mg/ml) to 1.95 (mg/ml). The activity strength of tetracycline against the extract was at 1:50. This revealed that Rambutan Binjai peel extract had great potency as antibacterial agent to MRSA. Dhaka Univ. J. Pharm. Sci. 17(2): 197-203, 2018 (December)

Saat ini, lebih dari setengah antibiotik yang digunakan di dunia merupakan kelompok β-laktam namun efektivitas klinis antibiotik tersebut kini terbatas karena resistensi antibiotik terhadap mikroorganisme penyebab penyakit infeksius. Beberapa mekanisme resistensi terhadap Enterobacteriaceae terutama disebabkan karena hidrolisis antibiotik oleh enzim spesifik, yang disebut dengan β-laktamase. Enzim β-laktamase menunjukkan kelompok besar enzim yang berbeda secara genetik dan fungsional yaitu extended-spectrum β-lactamase (ESBL) yang diketahui menimbulkan ancaman resistensi yang serius. Lokalisasi plasmid dari gen yang disandi terhadap distribusi enzim pada patogen meningkat setiap tahunnya. ESBL yang memiliki penyebaran yang luas dan relevan secara klinis adalah ESBL kelas A yaitu jenis Temoniera (TEM), Sulphydryl variable (SHV) dan Cefotaxime (CTX-M). Tujuan penulisan review ini adalah untuk mengkaji varian gen bla CTX-M yang banyak menyebabkan peningkatan resistensi antibiotik. Metode yang digunakan pada review ini yaitu penelusuran data berbasis Pubmed, Scopus dan Google Scholar tanpa pembatasan indeks faktor dengan kata kunci " bla CTX-M ", " Extended-spectrum β-lactamase ", dan " antibiotic resistance ". Kesimpulan dari review ini yaitu, ESBL jenis CTX-M telah menggantikan jenis TEM dan SHV secara dominan pada dekade terakhir. ESBL yang dihasilkan oleh Klebsiella pneumoniae diketahui muncul sebagai salah satu patogen nosokomial utama. Infeksi nosokomial yang disebabkan oleh CTX-M-15 pada Klebsiella pneumoniae mengalami peningkatan dalam beberapa tahun terakhir ini.

Graphical abstract, This study was performed to assess the safety of the oral acute and subchronic administration of Polypodium feei root extract through acute and subchronic studies in mice and rats, respectively. In the acute toxicity treatment, mice were grouped according to the dose (1000, 2000, 4000 and 5000 mg/kg, b.w) and were observed for mortality and toxicity signs for 14 days. In the subchronic treatment, there were six groups of rats (female and male), a control group, three test groups (100, 400, and 800 mg/kg, b.w), and two satellite groups (control satellite and satellite 800 mg/kg groups). The three test groups received the extract orally once daily for 90 days. No animals in the acute and subchronic treatment groups showed mortality and any signs of toxicity, with no significant difference in the body weight and organ index compared to the control. The LD50 of the extract was estimated to be higher than 5000 mg/kg, therefore regarded as practically non-toxic. The haematological profiles did not significantly change on exposure to the extract for 90 days, except the platelet count in the female animals which significantly decreased in animals treated with 400 and 800 mg/kg, returning to normal after 28 days of recovery. The 800 mg/kg dose significantly increased the urea concentration and induced lesions in the stomachs of female animals. However, this undesirable effect on the kidney was not strong, as the creatinine concentration remained in the normal limits, and the histopathological observations showed no alteration in the kidney tissues. No significant morphological alterations in organs were observed, only minor lesions in the liver. These results indicate that the P. feei root extract is safe for use as herbal medicine and recommended at doses lower than 400 mg/kg.

Context: Human Epidermal Growth Factor (hEGF) is a potential therapeutic protein that has been widely used as a healing agent for various chronic wounds. It induces the proliferation and metabolism of epithelial cells, regenerates skin cells, and validates skin elasticity. In the previous study, recombinant hEGF (rhEGF) had been successfully expressed extracellularly in Escherichia coli (E. coli) BL21 (DE3) using pectate lyase B (PelB) signal peptide. The previous study has shown that the medium concentration and the induction time influenced the production of rhEGF. Aims: Therefore, this study was conducted to optimize the induction time and medium concentration for rhEGF extracellular secretion then followed by scale-up production. Settings and Design: This experiment was carried out using E. coli BL21 (DE3) which contains pD881 plasmid that carries hEGF and PelB gene. Optimization design of induction time and medium concentration were obtained using Central Composite Design (CCD). Methods and Material: The method of study started by the rejuvenation of E. coli culture, extracellular secretion, and optimization in the flask scale then followed by scaled-up production with high-cell density culture in the fermenter. Statistical analysis used: The optimization was carried out using Response Surface Methodology (RSM) and multi regression analysis. Results: This work showed that the multiplication of 1.5-fold medium concentration with induction time 3h after the culture started gave the best result among another condition in this study. Additionally, the rhEGF production in the fermenter scale was identified by SDS-PAGE Tricine and quantified by ELISA, which showed 122.40 μg of the rhEGF per milliliter medium. Conclusions: In respect of the result, we conclude that the optimized condition of extracellular secretion was successfully obtained, and gives higher result before the previous study.

In this research, Escherichia coli BL21 (DE3) harboring an expression vector constructed with a rhamnose-inducible promoter and a pelB signal peptide was used as a host cell to produce MPT64 protein. The objective of this research was to figure out the optimum time of mpt64 gene expression through real-time monitoring of MPT64 protein production and distribution in host compartments. The mpt64 expression was regulated by the rhamnose presence at a concentration of 4 mM. The real-time isolated protein was monitored using polyacrylamide gel electrophoresis in denaturation condition. Based on real-time monitoring, the MPT64 protein (24 kDa) in the cytoplasm was optimum detected at 24 h after induction. For periplasmic fraction, the protein was detected at 4 h after induction but thinning at 15 h after induction. At 16 h after induction, the MPT64 protein band was found in the medium with increasing concentrations until 24 h. Thus, it can be concluded that the mpt64 gene expression was regulated in the presence of rhamnose as an inducer, and the proteins were shown to be translocated throughout the host cell compartment with different levels of protein accumulation at different times, according to the role of pelB as a signal peptide.

Pectin, a natural polysaccharide, has gained increasing attention due to not only its biomaterial properties but also its biomedical activities. One of the abundant sources of pectin is mangosteen (Garcinia mangostana L.) rind. In this study, we characterized the pectin from Indonesian mangosteen rind extract and evaluated its antioxidant activity. Pectin was extracted in acid condition and evaluated its physicochemical properties by fourier transform infrared (FTIR), powder X-ray diffractometer (PXRD), water content, ash content, equivalent weight, methoxyl level and of galacturonic acid content. Furthermore, the antioxidant activity of pectin was also observed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Pectin was successfully extracted from dry weight of Indonesian mangosten rind with yield about 1,16 ± 0,17%, fine powder, brownish and odorless. FTIR and PXRD results showed that pectin from mangosteen rind extract was amorphous and similar characteristic with a commercial pectin. The chemical properties of pectin such as water content, ash content, equivalent weight, methoxyl level and of galacturonic acid level were 9.85 ± 0.12%, 3.91 ± 0.17%, 6330.76 ± 220.43 g/mol, 2.86 ± 0.05% and 75.98 ± 0.88%, respectively. In addition, pectin showed an antioxidant activity with the IC50 about 161.94 ± 31.57 ppm. These results suggest that pectin from Indonesian mangosteen rind has the potential properties as biopolymers for biomedical applications with a low-methylated pectin and a moderate antioxidant activity.

Background Central obesity is a risk factor for metabolic syndrome. Subjects with central obesity have a higher risk of developing type 2 diabetes and cardiovascular disease. Many conditions affect the prevalence of central obesity, including energy expenditure, aging, proinflammatory conditions, and hormonal, genetic, and ethnic differences. Polymorphism of the APM1 gene, encoding the protein adiponectin, is closely related to metabolic syndrome. Adiponectin influences fatty acid oxidation and glucose intake in muscle. Therefore, variation in the APM1 gene is associated with diabetes and obesity. Purpose The aim of the present study was to investigate the correlation of the single-nucleotide polymorphism (SNP) of the APM1 SNP rs2241766 with body mass index (BMI) and lipid profiles in Indonesian (Bandung) subjects. Patients and methods Genotyping of the APM1 gene was performed using the Amplification Refractory Mutation System. Whole blood and serum of 54 subjects with central obesity (waist circumference [WC] ≥90 cm) and 53 healthy subjects (WC

Atenolol (ATE) is a cardio-selective β-blocker that is used in the treatment of hypertension over extended periods. However, ATE, like propranolol, has major potential for misuse as a performance-enhancing drug in several sports. Therefore, an efficient and selective separation method is required to detect and monitor the level of ATE in the body. This paper presents a molecularly imprinted polymer with specific and selective binding to ATE using precipitation polymerization. We show that when employed in an optimized molecular imprinted solid phase extraction (MI-SPE) protocol, recoveries of 93.65 ± 1.29% from spiked blood serum with excellent discrimination from other β-blocker drugs is possible. The methodology used in this study includes molecular modeling interaction between ATE and itaconic acid (ITA) as functional monomer, followed by determination of binding constants with spectrophotometry, synthesis of the polymer using precipitation polymerization and ending with characterization and application of polymers to extract ATE in serum. Docking analysis revealed a binding affinity between ATE and ITA of −2.0 kcal/mol with the formation of hydrogen bonding. The association constant between ATE and ITA was studied by UV titration in two different solvents, with evidence of an association constant 6.277 × 102 M−1 measured in acetonitrile: methanol (1:1). An optimized MI-SPE protocol was developed for the extraction of ATE from spiked blood serum, obtaining recoveries of 93.65% with excellent selectivity toward other β-blocker drugs.

Resistensi antibiotik telah menjadi permasalahan klinis yang utama terhadap kualitas hidup pasien infeksi saluran kemih (ISK) yang disebabkan oleh bakteri Escherichia coli. Perlu dilakukan pengujian zat aktif yang berasal dari bahan alam terhadap bakteri resisten tersebut. Ekstrak akar Bixa orella L. diketahui memiliki aktivitas terhadap E. coli yang sensitif. Oleh karena itu, penelitian ini bertujuan untuk mengetahui aktivitas akar Bixa orellana L. terhadap isolat klinis E. coli resisten. Metode yang digunakan adalah difusi agar dengan kertas cakram untuk uji aktivitas antibakteri dan metode pengenceran padat untuk penentuan KHM dan KBM. Hasil penelitian menunjukkan bahwa ekstrak akar kesumba keling (Bixa orellana L.) memiliki aktivitas antibakteri pada konsentrasi 50%, 40%, 30%, 20%, dan 10% dengan nilai KHM 1450 µg/mL untuk isolat pasien 1, dan 2970 µg/mL untuk isolat pasien 2. Nilai KBM 1480 µg/mL untuk isolat pasien 1, dan 3000 µg/mL untuk isolat pasien 2. Nilai kesetaraan aktivitas antibakteri gentamisin terhadap ekstrak etanol Bixa orella L. keling untuk E. coli resisten isolat klinis pasien 1 dan isolat klinis pasien 2 masing-masing sebesar 1 : 1,341x103 dan 1 : 1,254x103 .

Bakteri penyebab Infeksi saluran kemih (ISK) telah mengalami resistensi terhadap berbagai antibiotik. Pola resistensi bakteri tersebut perlu diperbaharui datanya untuk memastikan pemberian antibiotik yang tepat. Tujuan penelitian mengidentifikasi dan melihat pola resistensi bakteri penyebab ISK di Puskesmas Ibrahim Adjie Kota Bandung. Koloni bakteri yang berasal dari urin 9 pasien suspect ISK ditumbuhkan dan hanya sampel yang berasal dari 3 pasien memenuhi persyaratan terindikasi ISK karena adanya jumlah koloni di atas 105 cfu/ml. Selanjutnya dilakukan penentuan Gram bakteri, isolasi DNA total kromosom bakteri, amplifikasi gen pengkode 16S rRNA, sekuensing (penentuan urutan nukleotida) dan uji resistensi terhadap antibiotik golongan penisilin, penisilin+inhibitor β-lactamase, sefalosporin (generasi 1,2,3,4), fluorokuinolon, aminoglikosida dan karbapenem. Hasinya didapatkan tiga isolat (P1,P2,P3) yang teridentifikasi secara genetik Escherichia coli. Isolat P1 resisten terhadap penisilin, penisilin+inhibitor β-laktamse, sefalosporin generasi 1 dan fluorokuinolon. isolat P2 resisten terhadap penisilin. Isolat P3 resisten terhadap penisilin, penisilin+inhibitor β-laktamse, sefalosporin (generasi 1,2,3,4) dan fluorokuinolon. E. coli penyebab ISK telah mengalami resistensi terhadap antibiotik penisilin, penisilin+inhibitor β-lactamase, sefalosporin (generasi 1,2,3,4) dan fluorokuinolon. Aminoglikosida dan karbapenem masih memiliki aktivitas terhadap ketiga isolat E. coli resisten.Kata kunci: bakteri; ISK; isolasi; identifikasi; resistensi

Pakis tangkur merupakan tanaman obat tradisional yang sering dimanfaatkan oleh masyarakat untuk mengobati penyakit reumatik. Tanaman ini tumbuh di area pegunungan talaga bodas, Garut. Penelitian sebelumnya, melaporkan bahwa senyawa Shellegueain A yang terkandung dalam akar pakis tangkur yang berasal dari pegunungan Tangkuban Perahu efektif sebagai analgetik. Namun, dalam bentuk sediaan ekstrak yang secara umum merupakan sediaan yang paling banyak digunakan secara tradisional oleh masyarakat belum dilaporkan. Penelitian ini bertujuan untuk mengetahui aktivitas analgetik ekstrak etanol dosis 100, 200 dan 400 mg/kg serta fraksi n-heksan, etil asetat dan air dosis 50, 100 dan 200 mg/kg dengan metode geliat (siegmund) dan metode panas (hot plate). Hasilnya menunjukkan bahwa ekstrak etanol menghasilkan jumlah geliat hewan sebesar 85,5; 68,45 dan 77,35 kali dengan nilai persen proteksi 71,78; 77,4 dan 74,5 %. Sediaan fraksi n-heksan sebesar 90,45; 78,7 dan 85,95 kali (nilai persen proteksi 70,14; 74,03 dan 71,63 %); fraksi etil asetat sebesar 58,5; 102,5 dan 55,5 kali (nilai persen proteksi 80,05; 66,17 dan 79,68 %) serta fraksi air sebesar 205,2; 65,2 dan 60,5 kali (nilai persen proteksi sebesar 32,28; 78,48 dan 80,03 %). Aktivitas analgetik kuat hanya ditunjukkan oleh fraksi etil asetat dengan lama waktu respon 136,5; 147,5 dan 128,75 detik (nilai persen peningkatan 59,01; 55,4 dan 74,3 %) serta fraksi air sebesar 153; 144,75 dan 146 detik (persen peningkatan 57,39; 66,77 dan 71,47 %). Fraksi yang terbaik sebagai obat pereda nyeri adalah fraksi etil asetat terutama dosis 200 mg/kgbb.