Citric Acid Optimization by Candida tropicalis under Submerged Fermentation Conditions Using a Plackett-Burman Design

Citric acid production by fermentation is the most widely used way of obtaining it. The effects of some medium components were evaluated for Citric acid fermentation during the 1930s and 1940s.This work aimed to optimize citric acid by Candida tropicalis under submerged fermentation conditions using Plackett-Burman design. Some factors were tested as main variables affecting citric acid production using Plackett-Burman design. The results showed that incubation period of 7 days and pH 7; sodium acetate (10g/L), magnesium sulfate (1.5g/L), potassium phosphate (5g/L), ammonium chloride (3g/L), ferric sulfate(140mg/L), manganese sulfate (50 mg/L), zinc sulfate (80 mg/L), yeast extract (5.0g/L), glucose (150g/L), aeration ratio (75ml medium/ flask 250ml) were the most effective conditions for the highest yield of citric acid. The highest citric acid concentration was 30.0 g/L of the medium under the aforementioned conditions

Amelioration effect of 18β-Glycyrrhetinic acid on methylation inhibitors in hepatocarcinogenesis -induced by diethylnitrosamine

Aimsuppression of methylation inhibitors (epigenetic genes) in hepatocarcinogenesis induced by diethylnitrosamine using glycyrrhetinic acid.MethodIn the current work, we investigated the effect of sole GA combined with different agents such as doxorubicin (DOX) or probiotic bacteria (Lactobacillus rhamanosus) against hepatocarcinogenesis induced by diethylnitrosamine to improve efficiency. The genomic DNA was isolated from rats’ liver tissues to evaluate either methylation-sensitive or methylation-dependent resection enzymes. The methylation activity of the targeting genes DLC-1, TET-1, NF-kB, and STAT-3 was examined using specific primers and cleaved DNA products. Furthermore, flow cytometry was used to determine the protein expression profiles of DLC-1 and TET-1 in treated rats’ liver tissue.ResultsOur results demonstrated the activity of GA to reduce the methylation activity in TET-1 and DLC-1 by 33.6% and 78%, respectively. As compared with the positive control. Furthermore, the association of GA with DOX avoided the methylation activity by 88% and 91% for TET-1 and DLC-1, respectively, as compared with the positive control. Similarly, the combined use of GA with probiotics suppressed the methylation activity in the TET-1 and DLC-1 genes by 75% and 81% for TET-1 and DLC-1, respectively. Also, GA and its combination with bacteria attenuated the adverse effect in hepatocarcinogenesis rats by altering potential methylomic genes such as NF-kb and STAT3 genes by 76% and 83%, respectively.ConclusionGA has an ameliorative effect against methylation inhibitors in hepatocellular carcinoma (HCC) by decreasing the methylation activity genes

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

AMERICAN JOURNAL OF FOOD AND NUTRITION Production of a functional frozen yoghurt fortified with Omega-3 and Vitamin E

ABSTRACT Frozen yoghurt can be regarded as a healthy alternative to plain ice cream. This study investigates the isolation and identification of beneficial bacterial isolates from Laban Rayeb. Three isolates were selected and identified, namely Lactobacillus acidophilus r1, Lactobacillus acidophilus r2 and Latctococcus lactis subsp lactis r3. Also results showed that Laban Rayeb isolates had an inhibition and bactericidal effects on the growth of some pathogenic microorganisms as Staphylococcus aureus; E.coli ATCC 25922 and Bacillus subtilis NCIB3610. Antagonistic effect of S. aureus and E.coli indicated more pronounced inhibitory effect than Bacillus subtilis especially for Lactobacillus acidophilus r2. Then Low-fat ice cream mix was fermented with probiotic supplemented; omega-3 & vitamin E with traditional starter culture and evaluated for culture survival, composition, and sensory characteristics of frozen product. The developed frozen yoghurt formulae were as follows: Formula 1 with no probiotics, Formula 2 containing 3%w/w of Lactobacillus acidophilus r2 which has shown significant promise in all probiotics characteristics, and Formula 3 containing 3%w/w of probiotic strain (Lactobacillus acidophilus r2); with 2% ω-3 with vitamin E. The survival rate of probiotic in Formula 2 and Formula 3 after 4 weeks was higher than 10 6 CFU/ ml, the number regulated by FDA for the probiotic products. Formula 3 was containing 39.13±1.13 μg/ g alpha tocopherols after 4 weeks under freezing conditions. Sensory evaluation was carried out among 10 panelists, using the 9-point hedonic scale method for food acceptance. Formula 1, 2, and 3 obtained the mean score of 7.45, 7.98 and 8.01, respectively

Bio-control of Pseudomonas fluorescens in Domiati Cheese

<p>Concerning the antimicrobial activities of some probiotics bacteria Lactobacillus acidophilus P109, Lactobacillus plantarum P164, E. durans P174 and B. longum CHRS using an agar well-diffusion as In Vitro assay against selected isolates of Pseudomonas fluorescens, Bacillus cereus, Enterococcus fecalis and Staphylococcus aureus, indicated that B. longum CHRS appeared to have antimicrobial activity against these isolates. B. longum CHRS was injected with and without Ps. fluorescens in Domiati cheese during manufacturing as In Vivo experiment, results revealed that this strain of probiotic bacteria was reduced the count of Ps. fluorescens, while the chemical composition showed reduce production of soluble nitrogen, which has relation with the decomposition of protein as well as led to reduced volatile fatty acids, which refers to the decomposition of fat as a result of antimicrobial activity of B. longum against Ps. fluorescens</p&gt

Application of a New Simple Spectrophotometric Method to the Simultaneous Determination of Diclofenac Sodium and Diflunisal in Their Combined Dosage Form

ABSTRACT In this work, a simple and sensitive spectrophotometric method is presented for determination of the non-steriodal anti-inflammatory drugs; diclofenac sodium (DCL) and diflunisal (DIF) in their binary mixture without prior separation. The proposed method is based on the generation of ratio spectra of one compound using the other as the divisor followed by measurement of the peak-to-trough amplitudes between two selected wavelengths in the generated ratio spectra. For the determination of DCL, a standard solution of DIF 5 µg/mL was used as the divisor, and the peak-to-trough amplitudes between 251 and 291 nm were measured and correlated to the corresponding concentrations. Similarly, DCL 7.5 µg/mL was set as the divisor in DIF determination and the peak-to-trough amplitudes at the same wavelengths were recorded. The proposed method was found linear over the concentration ranges 5-50 and 1.5-30 µg/mL for DCL and DIF, respectively. The developed method was validated following the ICH guidelines and successfully applied to the determination of both drugs in various laboratory prepared mixtures. In addition, satisfactory results were obtained from analysis of the commercial pharmaceutical preparation (suppositories) with no significant statistical differences from a reference HPLC method

New simple spectrophotometric method for determination of the binary mixtures (atorvastatin calcium and ezetimibe; candesartan cilexetil and hydrochlorothiazide) in tablets

A new simple spectrophotometric method was developed for the determination of binary mixtures without prior separation. The method is based on the generation of ratio spectra of compound X by using a standard spectrum of compound Y as a divisor. The peak to trough amplitudes between two selected wavelengths in the ratio spectra are proportional to concentration of X without interference from Y. The method was demonstrated by determination of two drug combinations. The first consists of the two antihyperlipidemics: atorvastatin calcium (ATV) and ezetimibe (EZE), and the second comprises the antihypertensives: candesartan cilexetil (CAN) and hydrochlorothiazide (HCT). For mixture 1, ATV was determined using 10 μg/mL EZE as the divisor to generate the ratio spectra, and the peak to trough amplitudes between 231 and 276 nm were plotted against ATV concentration. Similarly, by using 10 μg/mL ATV as divisor, the peak to trough amplitudes between 231 and 276 nm were found proportional to EZE concentration. Calibration curves were linear in the range 2.5â40 μg/mL for both drugs. For mixture 2, divisor concentration was 7.5 μg/mL for both drugs. CAN was determined using its peak to trough amplitudes at 251 and 277 nm, while HCT was estimated using the amplitudes between 251 and 276 nm. The measured amplitudes were linearly correlated to concentration in the ranges 2.5â50 and 1â30 μg/mL for CAN and HCT, respectively. The proposed spectrophotometric method was validated and successfully applied for the assay of both drug combinations in several laboratory-prepared mixtures and commercial tablets. Keywords: Spectrophotometric analysis, Ratio spectra, Atorvastatin, Ezetimibe, Candesartan, Hydrochlorothiazid

Validated stability-indicating HPLC-DAD method for determination of the phosphodiesterase (PDE-4) inhibitor roflumilast

A comprehensive stability indicating HPLC with diode array detection method was developed for determination of the recently approved phosphodiesterase type 4 (PDE-4) inhibitor roflumilast (RFL). Effective chromatographic separation was achieved using Durashell C18 column (4.6 × 250 mm, 5 μm particle size) with isocratic elution of the mobile phase composed of 0.0065 M ammonium acetate pH 6.3, methanol and acetonitrile in the ratio of 30:35:35 (by volume). The mobile phase was pumped at a flow rate of 1.3 mL/min, and quantification of RFL was based on measuring its peak areas at 251 nm. RFL eluted at retention time 6.2 min. Analytical performance of the proposed HPLC procedure was thoroughly validated with respect to system suitability, linearity, range, precision, accuracy, specificity, robustness, detection and quantification limits. The linearity range was 2.5–200 μg/mL with correlation coefficient >0.9998. The drug was subjected to stress conditions of neutral, acidic and alkaline hydrolysis, oxidation, photolysis and thermal degradation. The proposed method proved to be stability-indicating by resolution of the drug from its forced degradation products. Moreover, specificity of the method was verified by resolution of drug from more than 20 pharmaceutical compounds of various medicinal categories. The validated HPLC method was successfully applied to the analysis of the cited drug in its tablet dosage form. The proposed method made use of DAD as a tool for peak identity and purity confirmation

Validated HPTLC method for the simultaneous determination of alfuzosin, terazosin, prazosin, doxazosin and finasteride in pharmaceutical formulations

Benign prostatic hyperplasia (BPH) is one of the most common chronic diseases affecting men and it increases in both incidence and prevalence with age. This work presents a simple, sensitive and fast generic high performance thin layer chromatographic (HPTLC) method for the simultaneous determination of five drugs prescribed for the treatment of BPH. These drugs include the α1-adrenergic blockers; alfuzosin hydrochloride (ALF), terazosin hydrochloride (TER), prazosin hydrochloride (PRZ) and doxazosin mesylate (DOX) in addition to the 5α-reductase inhibitor; finasteride (FIN). The cited drugs were separated on TLC-silica plates using a mobile phase composed of methylene chloride:n-hexane:methanol (8.8:0.3:0.9, by volume). Densitometric analysis was carried out at 254 nm for the α-blockers while FIN was measured at 220 nm. The five drugs were detected at Rf values of 0.26, 0.36, 0.45, 0.59 and 0.69 for ALF, TER, PRZ, DOX and FIN, respectively. The developed method was validated according to the International Conference on Harmonization (ICH) guidelines regarding; linearity, ranges, accuracy, precision, selectivity, robustness and limits of detection and quantification. The proposed method showed good linearity (r > 0.9990) in the ranges; 30–350, 30–350, 20–200, 30–350, 200–2000 ng/spot for the cited drugs, respectively. The applicability of the proposed method was verified through the analysis of laboratory-prepared mixtures and percentage recoveries between 98.27% and 101.97% were obtained. Commercial tablets were also analyzed by the developed methodology with no interference detected from the co-formulated excipients. The high sensitivity, simplicity and selectivity of the proposed method suggest its applicability for routine quality-control analysis purposes of any of the titled drugs in their pharmaceutical preparations

Evaluation of the General Organization of Veterinary Services control program of animal brucellosis in Egypt: An outbreak investigation of brucellosis in buffalo

Background and Aim: Brucellosis is a major constraint to livestock production in Egypt as well as many developing countries worldwide. Bovine brucellosis is an economically important disease with reproductive failure as a principal manifestation resulting in abortion, premature birth and decreased milk production in females, and orchitis and epididymitis in males. In spite of the efforts of Egyptian veterinary services to overcome brucellosis, the disease is still prevalent in both animals and humans and represents one of the most important public health hazards in Egypt. The aim of the present work was to investigate the efficacy of the control program implemented by the General Organization of Veterinary Services in Brucella infected buffalo farm on serological, molecular, cultural, and histopathological basis. Brucella melitensis biovar 3 was recovered from 6 buffalo-cows. Materials and Methods: Blood samples were collected from a total of 750 non-vaccinated lactating buffalo-cows. These animals were proved positive for Brucella by the Egyptian brucellosis national program. Sera were tested using buffered acidified plate antigen test and rose Bengal test as screening tests and complement fixation test as a confirmatory test. Positive animals were separated for slaughtering under the supervision of the Egyptian veterinary authorities. Remaining animals were tested every 3 weeks with slaughtering of positive cases and this continued until the remaining animals revealed three successive negative serological tests. Different lymph nodes (prescapular, prefemoral, mediastinal, retropharyngeal, and supramammary) were collected from 11 Brucella seropositive buffalo-cows slaughtered after being confirmed serologically as Brucella infected cases. Samples were collected and processed for bacterial isolation and nucleic acid detection using polymerase chain reaction (PCR). Parts of these specimens were fixed in 10% neutral buffered formalin for 48 h then processed by paraffin embedding technique. Results: "Test and slaughter" policy was applied on Brucella infected dairy buffalo farm. The program continued for 6 months with slaughtering of positive cases until the herd was proved Brucella free. B. melitensis biovar 3 could be recovered from six buffalo-cows. Universal PCR confirmed Brucella on genus level and Bruce-ladder multiplex, PCR confirmed the presence of B. melitensis on the species level. Histopathological examination of Brucella-infected lymph nodes revealed massive rarified and depleted lymphoid areas of both sub-capsular and deep cortical lymphoid follicles, macrophage cells granulomatous reaction, as well as fat, infiltrates, and chronic vasculitis. The chronic nature of Brucella lesions has been confirmed in this study as indicated by the chronic vasculitis and collagen deposition. Conclusion: Freedom status from brucellosis in this study required 6 months which are considered long time allowing the spread of infection to other localities especially under unhygienic conditions, husbandry system favoring mixed populations of different ages, sex, aborted and pregnant, and lack of controlled movement of animals. Therefore, effective control of animal brucellosis requires surveillance to identify infected animal herds, elimination of the reservoirs, and vaccination of young heifers. B. melitensis biovar 3 is the cause of the Brucella outbreak in buffalo which still remains the prevalent type of Brucella in Egypt. The disease runs a chronic course allowing further spread of infection