Optimization of dextran production from Leuconostoc strains isolated from different dairy products from Ouargla region Algeria

Leuconostoc (Ln) sp. belongs to a group of lactic acid bacteria, which has the capacity to produce dextran (an exopolysaccharides) in the presence of su-crose. dextran is industrially important, it was the first microbial exopolysac-charide affirmed for commercial use. This study aimed to optimize the pro-duction of the synthesized dextran by Ln strains species isolated from differ-ent dairy products. Morphological, cultural, physiological and biochemical characteristics were employed to identify 23 isolated strains. We have identi-fied the species: Ln. gelidum, Ln. carnosum, Ln. citreum, Ln. fallax, Ln. mesen-teroides subsp mesenteroides, Ln. mesenteroides subsp dextranicum, Ln. mesenteroides subsp cremoris. 20 strains had the capacity to produce dex-tran from sucrose. The precipitation and quantification of EPS on MRSs (Mark rogosa et sharpe sucrose) medium revealed a difference between the strains, by the total sugars assay method, the amount of EPS varied between 0.63 ± 0.19 and 2.41 ± 0.17 g / L of strains LnF70 and LnC1 (isolated from goat's milk), respectively. The dextran production from MRSs medium was better than from liquid MSE. The optimization of production on MRSs medi-um with different concentration of glucose, yeast extract and sucrose showed that the strains had good production with a concentration of 2% glucose, 0.3% yeast extract and 10% sucrose.

In Vitro Assessment of the Cercaricidal Activity of Sida acuta Burm. F. and Sida rhombifolia Linn. (Malvaceae) Hydroethanolic Extracts, Cytotoxicity, and Phytochemical Studies

Despite the global efforts, schistosomiasis remains a public health problem in several tropical and subtropical countries. One of the major challenges in the fight against schistosomiasis is the interruption of the parasite life cycle. Here, we evaluated the anticercarial, cytotoxicity, and phytochemical profiles of Sida acuta (HESa) and Sida rhombifolia (HESr) hydroethanolic extracts (Malvaceae). Schistosoma mansoni cercaria was collected from fifteen Biomphalaria pfeifferi-infected snails. Twenty-five cercariae were incubated in duplicate with different concentrations (31.25–1,000 μg/mL) of HESa or HESr. The cercaria viability was monitored at 30 min time intervals for 150 min, and the concentration-response curve of each plant extract was used to determine their respective lethal concentration 50 (LC50). Additionally, the cytotoxicity profile of each plant extract was evaluated on the Hepa 1–6 cell line at a concentration range of 15.625–1,000 µg/mL using the WST-8 assay method and its inhibitory concentration 50 (IC50) was calculated. Moreover, phytochemical characterization of each plant extract was carried out by HPLC-MS. Both extracts exhibited cercaricidal activity in a time- and concentration-dependent manner. At 30 min time point, HESa (LC50 = 28.41 ± 3.5 µg/mL) was more effective than HESr (LC50 = 172.42 ± 26.16 µg/mL) in killing S. mansoni cercariae. Regarding the cytotoxicity effect of both extracts, the IC50 of HESa (IC50 = 109.67 µg/mL) was lower than that of HESr (IC50 = 888.79 µg/mL). The selectivity index was 3.86 and 5.15 for HESa and HESr, respectively. Fifteen compounds were identified from HESa and HESr after HPLC-MS analysis. N-Feruloyltyramine, a polyphenol, and thamnosmonin, a coumarin, were identified in both extracts. HESa and HESr displayed cercaricidal activity and were not toxic on Hepa 1–6 cell line. Based on the selectivity index of these extracts, S. rhombifolia extract could be more effective on S. mansoni cercariae than S. acuta extract. This study could provide baseline information for further investigations aiming to develop plant-based alternative drugs against S. mansoni.

METHOD VALIDATION OF SIMVASTATIN IN PCL-PEG-PCL TRIBLOCK COPOLYMER MICELLES USING UV-VIS SPECTROPHOTOMETRIC FOR SOLUBILITY ENHANCEMENT ASSAY

Objective: This study aims to increase the solubility of simvastatin (SIM), a hydrophobic drug, by incorporating it into PCL-PEG-PCL triblock copolymer micelles and validating the assay method used, namely Uv-Vis spectrophotometric. Methods: The shake flask method was used to determine the increase in solubility experienced by SIM after being incorporated into the micellar system. The values ​​of maximum wavelength (λmax), linearity, LOD, LOQ, accuracy, and precision were used as parameters measured to assess the validity of the assay method used. Results: The results showed that PCL-PEG-PCL triblock copolymer micelles could increase SIM solubility by 9.7 times (89.49±5.75 µg/ml) compared to SIM without modification (9.19±0.24 µg/ml). The validation results show the λmax value of 239 nm, a linear calibration curve with an R-value of 0.9994, LOD and LOQ of 0.33 µg/ml and 1.00 µg/ml, accurate measurement with recovery at concentrations of 80%, 100%, and 120% were 102.93±1.32%, 100.78±0.40%, and 104.58±0.79% and also had good precision ​​with RSD<2%. Conclusion: The PCL-PEG-PCL triblock copolymer micelles can increase SIM solubility and the Uv-Vis spectrophotometric method has been validated successfully for the quantitative analysis of SIM in PCL-PEG-PCL triblock copolymer micelles.

Applying Four-Step Characteristic Ion Filtering with HPLC-Q-Exactive MS/MS Spectrometer Approach for Rapid Compound Structures Characterization and Major Representative Components Quantification in Modified Tabusen-2 Decoction

Modified Tabusen-2 decoction (MTBD) is traditional Chinese Mongolia medicine, mainly used to treat osteoporosis. However, the precise material basis of this prescription is not yet fully elucidated. Herein, we establish an HPLC-Q-Exactive MS/MS spectrometer method with four-step characteristic ion filtering (FSCIF) strategy to quickly and effectively identify the structural features of MTBD and determine the representative compounds content. The FSCIF strategy included database establishment, characteristic ions summarization, neutral loss fragments screening, and secondary mass spectrum fragment matching four steps. By using this strategy, a total of 143 compounds were unambiguously or tentatively annotated, including 5 compounds which were first reported in MTBD. Nineteen representative components were simultaneously quantified with the HPLC-Q-Exactive MS/MS spectrometer, and it is suitable for eight batches of MTBD. Methodology analysis showed that the assay method had good repeatability, accuracy, and stability. The method established above was successfully applied to assess the quality of MTBD extracts. Collectively, our findings enhance our molecular understanding of the MTBD formulation and will allow us to control its quality in a better way. At the same time, this study can promote the development and utilization of ethnic medicine.

Vitamin B12 deficiency is linked with dyslipidemia in gestational diabetes mellitus.

Objective: To determine the vitamin B12 deficiency and dyslipidemia in Gestational Diabetes mellitus (GDM) diagnosed pregnant women. Study Design: Observational study. Setting: Department of Biochemistry and Gynecology/Obstetrics LUMHS Hospital Jamshoro. Period: January 2018 to December 2018. Material & Methods: A sample of 216 diagnosed GDM pregnant women was selected according to study criteria. Venous blood samples were centrifuged to separate sera; that were used for the estimation of (hexokinase method), blood lipids and Vitamin B12 (ECLIA assay method) by Cobas chemistry analyzer. Statistical SPSS software 21.0 (IBM, Inc USA) was used for study research variables at 95% CI (P ≤ 0.05). Results: Age of GDM cases was 36.12±9.5 years. Mean+/-SD vitamin B12 level was noted 154.7±81.7 ng/mL (P=0.0001). Serum cholesterol, triglycerides and LDLc were elevated and HDLc was low in GDM cases (P=0.0001). Of 216 GDM cases, vitamin B12 deficiency was present in 152 (70.3%) (P=0.0001) and dyslipidemia in 50 (23.1%) (X2=452.0) (P=0.0001). Vitamin B12 shows inverse correlation with RBG (r= -0.41, P=0.005), CHOL (r= -0.25, P=0.024), TAG (r= -0.81, P=0.0001), LDLc (r= -0.797, P=0.0001) and positive correlation with HDLc (r= 0.76, P=0.0001). Conclusion: The present study finds vitamin B12 deficiency in 152 (70.3%) and dyslipidemia in 50 (23.1%). Vitamin B12 deficient GDM women show high cholesterol, triglycerides, LDLc and low HDLc. Hence, it is concluded, the vitamin B12 deficiency is linked with dyslipidemia in Gestational Diabetes mellitus.

Polysaccharide hydrolyzing enzyme activity of bacteria, native to Apis florea gut

Introduction and Aim: Apis florea commonly known as “dwarf honey bee” harbors enormous gut bacteria that can digest complex carbohydrates and other food components. In this regard, the present investigation was focused on analyzing the polysaccharide degrading ability of bacteria isolated from the gut of honeybee, for their possible application in nutraceutical and pharmaceutical industries. Materials and Methods: Nine bacterial isolates were screened for carbohydrate degrading enzymes viz., amylase, pectinase, cellulase, tannase and laccase, using respective substrate by plate assay method. Further activities of amylase and pectinase were measured quantitatively by dinitrosalicylic acid (DNS) method. Results: All the nine selected isolates exhibited amylase and pectinase activities. However, only two isolates exhibited lignolytic and cellulolytic activity. None of the isolates showed tannin degradation. Maximum amylase activity (4.95 U/mg) was observed in Bacillus halotolerans af-M9 followed by Klebsiella oxytoca af-G4 (4.62 U/mg). With respect to pectinase activity Klebsiella pneumoniae af-E17 displayed higher activity (0.24 U/mg) followed by Klebsiella oxytoca af-G4 (0.20 U/mg). Conclusion: Habitat-specific innovations are being explored for novel compounds for therapeutic applications. This study throws a light on selection of carbohydrate degrading bacteria from a new source i.e., GUT of honeybee.

ATOPIC DERMATITIS: IMPROVING EARLY DETECTION AND PREVENTION MEASURES IN THE COMMUNITY SETTING

Atopic dermatitis is a chronic inflammation of the skin characterized by disruption of the skin barrier and aberrations in the immune response. In general, atopic dermatitis can be affected by various complex interactions including genetics, diet, and stress. The lack of public attention to atopic dermatitis which often affects children is one of the factors for the increasing prevalence of atopic dermatitis. The purpose of this community service is to help the community understand the importance of early detection and allergy prevention efforts. Community service is carried out by holding online seminars that can be followed by the general public as well as direct consultations and discussions with pediatricians for allergy patients at the Department of Pediatrics at Dr. RSUD Dr. Saiful Anwar Malang. The total Immunoglobulin E level of the respondents was measured using the enzyme-linked immunosorbent assay method. The severity of the respondents was determined by calculating the SCORAD index. The results of early detection showed that 100% of respondents had intrinsic atopic dermatitis, 90% with mild severity and 10% with moderate severity. After being checked for allergies, the respondents received the appropriate supplements. Conclusion: By holding this community service activity, ordinary people become more aware of the importance of treating allergic diseases as early as possible because the incidence of atopic dermatitis can be reduced and the quality of life of patients can be improved. This outreach activity must be carried out continuously in the following years.

DISAPPEARANCE OF ANTIBODIES IN COVID-19 MYTHS ORREALITY

Objective: To explore the disappearance of neutralizing antibodies from patients, their myths, and facts. Study Design: Cross-sectional study. Place and Duration of Study: Combined Military Hospital Multan Pakistan, from Jul 2021 to Aug 2021. Methodology: A total of 100 blood samples were collected from 100 COVID-19 patients. These 100 patients were followed up for a period of 3 months. Antibodies were determined with the modified neutralization assay method and enzyme-linked immuno-sorbent assay (ELISA). Results: The antibody level by NA and ELISA peaked on days 30-35 then decreased slightly. In multivariate analysis, patients aged 25-35, 36-56, and 57-84 years had a higher neutralizing antibody level than those aged 10-21 years. The patient with the worst clinical manifestation had a higher neutralizing antibody titer. In serum samples, IgG was undetectable at 18.3% and 11% and the geographical mean reciprocal titers dropped from 244 at 3-month period and neutralizing antibodies, the geographical mean reciprocal titers dropped from 874 at 3 months. Conclusion: All COVID-19 patients were seropositive and significantly neutralizing antibody response. Neutralizing antibody levels depend on the time after the onset of symptoms, age, and severity of the disease.

Effect of Imidocarb Application on Oxidative DNA Damage Caused by Anaplasmosis

This study was aimed to evaluate DNA fragmentation by using Comet assay in naturally infected sheep with Anaplasmosis before and after treatment with the Comet method, which shows DNA damage specifically. In the study, blood samples were collected from 10 Anaplosmosis infected and 10 healthy sheep. The anaplosmosis was diagnosed by clinical signs and symptoms. The infection was confirmed by Giemsa staining. The blood was collected from control group and infected group before and after the treatment, from the vena jugularis with the appropriate method. The DNA fragmentation was checked by using the Comet assay of blood cells. The data were analysed throught ANNOVA one-way. The result showed higher DNA fragmentation in sick animals diagnosed with anaplasmosis; tail length and tail moment values were found to be statistically significantly higher than the control group. When the data obtained after imidocarb (IMD) application were compared with obtained during the disease, a decreased DNA damage and tail moment was determined, however, these values higher than control. In this study, DNA damage and the extent of this damage were investigated by the Comet assay method using a healthy control group before and after treatment in animals with Anaplasmosis. When the findings obtained from the study were evaluated, it was seen that Anaplasma agents caused DNA damage and with the imidocarb application given for treatment, DNA damage was reduced and results close to healthy individuals were obtained.