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Background: Management of ever growing multidrug resistant (MDR) bacteria becomes one of the biggest threats to public health worldwide. The situation is worsening due to lack of new generation antimicrobials in the arsenal of the clinicians. Development of new alternatives to the conventional antimicrobial agents is the need of the hour to control the menace of AMR. Plants based products are attractive alternatives with proven efficacy but needs scientific investigation to explore their potential antimicrobial, antibiofilm and antiquorum sensing activities against major bacterial pathogens of human and animals. The present study was conducted to explore the antimicrobial, antibiofilm and antiquorum sensing activity of aqueous and methanol extracts of leaf, flower, fruit and stem of the Himalayan yellow raspberry (Rubus ellipticus) against clinical isolates of Staphylococcus aureus and Escherichia coli. Methods: E. coli and S. aureus were isolated and identified from diarrhoeic pigs and poultry and mastitic milk of cattle of Mizoram, respectively. Leaf, flower, fruit and stem/bark of R. ellipticus were collected from Mizoram and extracted by methanol and aqueous solvents. The antimicrobial activity and MIC was determined by well diffusion method and 96 wells microtiter plate method, respectively. Antibiofilm activity of plant extracts was determined in 96 well tissue culture plate. Antiquorum sensing activity was determined by disc diffusion method. Result: Methanol leaf extract exhibited antimicrobial activity against S. aureus with 19 mm and 7 mm zone of inhibition at 200 mg/mL and 12.5 mg/mL, respectively. Methanol fruit extract also showed antimicrobial activity against S. aureus only. Highest and lowest activities were observed at 200 mg/mL and 25 mg/mL concentrations with 15 mm and 7 mm zone of inhibition, respectively. No antimicrobial activities by either of the extracts were observed against E. coli isolates. The MIC of R. ellipticus methanol leaf and fruit extracts against S. aureus was 0.203125 mg/mL and 0.8125 mg/mL, respectively. The methanol leaf (86.60%) and stem (85.60%) extracts of R. ellipticus showed significant antibiofilm activity against S. aureus isolates, whereas methanol fruit (89.20%) extracts exhibited antibiofilm activity against E. coli isolates at the concentration of 0.05 mg/mL. Significant antiquorum sensing (QS) activities was exhibited by the methanol leaf extract of R. ellipticus at 200 mg/mL concentration against E. coli. This is the first ever report on antibiofilm and anti QS activities of the R. ellipticus plant extracts against E. coli and S. aureus bacteria. [ABSTRACT FROM AUTHOR]

Background: Development and persistence of multidrug resistant (MDR) bacteria is considered to be one of the biggest threats to public health worldwide. Development of new antimicrobial agents and alternatives to the conventional antimicrobial agents to control the menace of AMR is the need of the hour. Plants based products can be effectively explored as potential antimicrobial, antibiofilm and antiquorum sensing agents against major bacterial pathogens of human and animals. This present study was conducted to explore the antimicrobial, antibiofilm and antiquorum sensing activity of aqueous and methanol extracts of leaf, flower, fruit and stem of Melastoma malabathricum against clinical isolates of Staphylococcus aureus and Escherichia coli. Methods: E. coli and S. aureus were isolated and identified from diarrhoeic pigs and poultry and mastitic milk of cattle of Mizoram, respectively. Leaf, flower, fruit and stem of M. malabathricum were collected from Mizoram and extracted by methanol and aqueous solvents. The antimicrobial activity and MIC was determined by using well diffusion method and 96 wells microtiter plate method, respectively. Antibiofilm activity of plant extracts was determined in 96 well tissue culture plate. Antiquorum sensing activity was determined by disc diffusion method. Result: Methanol leaf extract exhibited antimicrobial activity against E. coli but not against S. aureus with 18 mm and 6 mm zone of inhibition at 200 mg/mL and 12.5 mg/mL, respectively. Methanol flower extract showed antimicrobial activity against S. aureus but not against E. coli with 14 mm and 6 mm zone of inhibitions at 200 mg/ml and 12.5 mg/mL, respectively. Similarly, the aqueous leaf extract showed antimicrobial activity against S. aureus but not against E. coli with 12 mm and 6 mm zone of inhibition at 200 mg/mL and 100 mg/mL, respectively. The MIC of M. malabathricum methanol leaf extract against E. coli was 3.125 mg/mL, whereas the MIC value of methanol flower and leaf extracts was 6.25 mg/mL against S. aureus. Antibiofilm activity of M. malabathricum methanol leaf, methanol flower and aqueous leaf extracts was recorded only against S. aureus isolates with maximum inhibition at 0.05 mg/mL concentration. Good antiquorum sensing activities was exhibited by the M. malabathricum methanol leaf, methanol flower and aqueous leaf extracts against S. aureus isolates at 200 mg/mL concentration. [ABSTRACT FROM AUTHOR]

The octoploid-cultivated strawberry variety Benihope (Fragaria × ananassa Duch cv. Benihope) is an important commercial plant. It is highly susceptible to different diseases, which ultimately leads to a reduction in yield. Gene-editing methods, such as CRISPR/Cas9, demonstrate potential for improving disease resistance in the strawberry cv. Benihope. Establishing a plant regeneration system suitable for CRISPR/Cas9 gene editing is crucial for obtaining transgenic plants on a large scale. This research established a callus induction and plant regeneration system for Agrobacterium-mediated CRISPR/Cas9 gene editing in strawberry cv. Benihope by evaluating multiple types of explants and various plant growth regulators throughout the entire tissue culture process. The results showed that the efficiency of callus induction is strongly influenced by the type of explant and is highly sensitive to the combination of plant growth regulators. Among the different plant growth regulators employed, thidiazuron (TDZ), in combination with 2,4-dichlorophenoxyacetic acid (2,4-D), effectively induced callus formation and plant regeneration from explants derived from nutrient tissues such as runner tips and crowns. In addition, the regeneration experiment demonstrated that the addition of polyvinylpyrrolidone (PVPP) to the shoot regeneration medium could inhibit tissue browning. The gene-edited plants in which some or all of the Fvb7-1, Fvb7-2, Fvb7-3, and Fvb7-4 genes in the MLO (Mildew resistance Locus O) gene family were knocked out by CRISPR/Cas9 system were obtained by applying the plant regeneration system developed in this study. [ABSTRACT FROM AUTHOR]

The development of cross-protective vaccines against the zoonotic swine influenza A virus (swIAV), a potential pandemic-causing agent, continues to be an urgent global health concern. Commercially available vaccines provide suboptimal cross-protection against circulating subtypes of swIAV, which can lead to worldwide economic losses and poor zoonosis deterrence. The limited efficacy of current swIAV vaccines demands innovative strategies for the development of next-generation vaccines. Considering that intramuscular injection is the standard route of vaccine administration in both human and veterinary medicine, the exploration of alternative strategies, such as intradermal vaccination, presents a promising avenue for vaccinology. This investigation demonstrates the first evaluation of a direct comparison between a commercially available multivalent swIAV vaccine and monovalent whole inactivated H1N2 swine influenza vaccine, delivered by intradermal, intranasal, and intramuscular routes. The monovalent vaccines were adjuvanted with NanoST, a cationic phytoglycogen-based nanoparticle that is combined with the STING agonist ADU-S100. Upon heterologous challenge, intradermal vaccination generated a stronger cross-reactive nasal and serum antibody response in pigs compared with intranasal and intramuscular vaccination. Antibodies induced by intradermal immunization also had higher avidity compared with the other routes of vaccination. Bone marrow from intradermally and intramuscularly immunized pigs had both IgG and IgA virus-specific antibody-secreting cells. These studies reveal that NanoST is a promising adjuvant system for the intradermal administration of STING-targeted influenza vaccines. [ABSTRACT FROM AUTHOR]

Cellulase is a biocatalyst that hydrolyzes cellulosic biomass and is considered a major group of industrial enzymes for its applications. Extensive work has been done on microbial cellulase but fungi are considered a novel strain for their maximum cellulase production. Production cost and novel microbial strains are major challenges for its improvement where cheap agro wastes can be essential sources of cellulose as substrates. The researcher searches for more cellulolytic microbes from natural sources but the production level of isolated strains is comparatively low. So genetic modification or mutation can be employed for large-scale cellulase production before optimization. After genetic modification than in silico molecular modeling can be evaluated for substrate molecule's binding affinity. In this review, we focus not only on the conventional methods of cellulase production but also on modern biotechnological approaches applied to cellulase production by a sequential study on common cellulase-producing microbes, modified microbes, culture media, carbon sources, substrate pretreatment process, and the importance of optimum pH and temperature on fermentation. In this review, we also compare different cellulase activity determination methods. As a result, this review provides insights into the interrelationship between the characteristics of optimizing different culture conditions, genetic modification, and in silico enzyme modeling for the production of cellulase enzymes, which may aid in the advancement of large-scale integrated enzyme manufacturing of substrate-specific enzymes. [ABSTRACT FROM AUTHOR]

Without adaptive immunity, invertebrates have evolved innate immune systems that react to antigens on the surfaces of pathogens. These defense mechanisms are included in horseshoe crab hemocytes' cellular responses to pathogens. Secretory granules, large (L) and small (S), are found on hemocytes. Once the invasion of pathogens is present, these granules release their contents through exocytosis. Recent data in biochemistry and immunology on the granular constituents of granule-specific proteins are stored in large and small granules which are involved in the cell-mediated immune response. L-granules contain most clotting proteins, which are necessary for hemolymph coagulation. They also include tachylectins; protease inhibitors, such as cystatin and serpins; and anti-lipopolysaccharide (LPS) factors, which bind to LPS and agglutinate bacteria. Big defensin, tachycitin, tachystatin, and tachyplesins are some of the essential cysteine-rich proteins in S-granules. These granules also contain tachycitin and tachystatins, which can agglutinate bacteria. These proteins in granules and hemolymph act synergistically to fight infections. These biomolecules are antimicrobial and antibacterial, enabling them to be drug resistant. This review is aimed at explaining the biomolecules identified in the horseshoe crab's hemolymph and their application scopes in the pharmaceutical and biotechnology sectors. [ABSTRACT FROM AUTHOR]

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoglobulin G (IgG) detection can be an effective complementary tool to the reverse transcription–polymerase chain reaction (RT-PCR) test in estimating the true burden of coronavirus diseases 2019 (COVID-19) and can serve as baseline data, especially after the roll-out of vaccines against SARS-CoV-2. In this study, we aim to determine the seropositivity of SARS-CoV-2 IgG among people in Dhaka, Bangladesh. Volunteers, mostly asymptomatic people from Dhaka, were enrolled between October 2020 and February 2021. After obtaining participants' signed consents, blood samples were tested for SARS-CoV-2 IgG antibody, following the standard protocol of testing within 72 hours of collection. SARS-CoV-2 IgG was positive in 42% (101/239) of the cases. No difference was observed in terms of IgG positivity and IgG levels when stratified by age, gender, and blood group. However, RT-PCR-positive cases presented higher IgG levels compared to RT-PCR-negative/RT-PCR-not performed cases. SARS-CoV-2 IgG was found in 31% (32/102) and 28% (19/67) of RT-PCR-negative and RT-PCR-not performed cases, respectively. For RT-PCR-positive but SARS-CoV-2 IgG-negative cases (n = 13), the average time gap between the RT-PCR and SARS-CoV-2 IgG tests of six months indicates a gradual reduction of IgG. Eight cases for which samples were tested at two time points, three months apart, showed presented a decline in IgG levels with time (median IgG index of 2.55 in the first sample versus 1.22 in the second sample). Our findings reveal that several mild/asymptomatic cases that were RT-PCR-negative/not tested exist in the community, and IgG levels reduce in the human body over time. [ABSTRACT FROM AUTHOR]

Aims/Introduction: Peroxisome proliferator‐activated receptor (PPAR)‐γ2 is a transcription factor crucial for regulating adipogenesis and glucose/lipid metabolism, and synthetic PPARγ ligands, such as thiazolidinediones, are effective oral medication for type 2 diabetes. Sirtuin 7 (SIRT7), a nicotinamide adenine dinucleotide‐dependent deacetylase, also controls metabolism. However, it is not known whether SIRT7 regulates the function of PPARγ2 by its deacetylation. Materials and Methods: Physical interaction between SIRT7 and PPARγ2, the effect of SIRT7 on PPARγ2 acetylation, and the deacetylation residue targeted by SIRT7 were investigated. The effects of PPARγ2 K382 acetylation on lipid accumulation, gene expression in C3H10T1/2 cell‐derived adipocytes, and ligand‐dependent transactivation activity were also evaluated. Results: We demonstrated that SIRT7 binds to PPARγ2 and deacetylates PPARγ2 at K382. C3H10T1/2‐derived adipocytes expressing PPARγ2K382Q (a mimic of acetylated K) accumulated much less fat than adipocytes expressing wild‐type PPARγ2 or PPARγ2K382R (a mimic of nonacetylated K). Global gene expression analysis of adipocytes expressing PPARγ2K382Q revealed that K382Q caused the dysregulation of a set of genes involved in lipogenesis, including Srebp1c, Acaca, Fasn, and Scd1. The rosiglitazone‐dependent transcriptional activity of PPARγ2K382Q was reduced compared with that of PPARγ2K382R. Conclusion: Our findings indicate that SIRT7‐dependent PPARγ2 deacetylation at K382 controls lipogenesis in adipocytes. [ABSTRACT FROM AUTHOR]

Lysosomes are the main degradative organelles of cells and involved in a variety of processes including the recycling of macromolecules, storage of compounds, and metabolic signaling. Despite an increasing interest in the proteomic analysis of lysosomes, no systematic study of sample preparation protocols for lysosome enriched fractions has been performed to date. In the current study, we used samples enriched for lysosomes by paramagnetic nanoparticles and systematically evaluated experimental parameters for the analysis of the lysosomal proteome. This includes different approaches for the concentration of lysosome-containing fractions; desalting of samples by solid phase extraction; fractionation of peptide samples; and different gradient lengths for LC-MS/MS analyses of unfractionated samples by data dependent and data independent acquisition. Furthermore, we evaluated four different digestion methods including filter aided sample preparation (FASP), in-gel digestion, and in-solution digestion using either RapiGest or urea. Using the combined data, we generated a benchmark lysosomal proteome data set for mouse embryonic fibroblasts as well as a spectral library for the analysis of lysosomes by data independent acquisition. Measurement(s) Proteome Technology Type(s) liquid chromatography-tandem mass spectrometry Sample Characteristic - Organism Mus musculus Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.11770893 [ABSTRACT FROM AUTHOR]

With increasing health awareness, the demand for concentrated proteins from plant sources has been rising for direct consumption or to use as an ingredient in food formulation. The characteristic information of any component is prerequisite for its efficient usage in food preparation. This study was undertaken to isolate the protein from jackfruit seeds and characterizing the physicochemical and functional properties of the isolated protein. The protein fraction from jackfruit seed flour was isolated using pH treatments and centrifugation process. The isolated protein was converted into powder form by a vacuum drying and grinding method. Total protein content in the isolate was determined by the Kjeldahl method. The functional properties such as solubility and gelling capacity and secondary structural elements of jackfruit seed protein isolate (JSPI) were studied. The crude JSPI contained 76.89% protein with 58.44% solubility in aquatic solvent. The conformational study by Fouriertransform infrared spectroscopy (FTIR) indicated that the ß -sheet is the dominant secondary structure of JSPI that contained 50.28% ß -sheet, 21.71% a -helix, 8.86% ß -turn, and 19.15% unordered structure. The least gelation concentration of JSPI dissolved in 1.0 M NaCl solution was 12%. The pH of the solvent significantly affected the emulsifying and foaming properties (p<0.05). Based on the observed structural and functional features, JSPI has prospects to be used as a supplementary ingredient in future food formulations. [ABSTRACT FROM AUTHOR]

To examine rates of Shigella infections in household contacts of pediatric shigellosis patients, we followed contacts and controls prospectively for 1 week after the index patient obtained care. Household contacts of patients were 44 times more likely to develop a Shigella infection than were control contacts (odds ratio 44.7, 95% CI 5.5-361.6); 29 (94%) household contacts of shigellosis patients were infected with the same species and serotype as the index patient's. Pulsed-field gel electrophoresis showed that 14 (88%) of 16 with infected contacts had strains that were indistinguishable from or closely related to the index patient's strain. Latrine area fly counts were higher in patient households compared with control households, and 2 patient household water samples were positive for Shigella. We show high susceptibility of household contacts of shigellosis patients to Shigella infections and found environmental risk factors to be targeted in future interventions. [ABSTRACT FROM AUTHOR]

Objective: To investigate the prevalence and mechanisms of fluoroquinolone resistance in Shigella species isolated in Bangladesh and to compare with similar strains isolated in China. Methods: A total of 3789 Shigella isolates collected from Clinical Microbiology Laboratory of icddr,b, during 2004–2010 were analyzed for antibiotic susceptibility. Analysis of plasmids, plasmid-mediated quinolone-resistance genes, PFGE, and sequencing of genes of the quinolone-resistance-determining regions (QRDR) were conducted in representative strains isolated in Bangladesh and compared with strains isolated in Zhengding, China. In addition, the role of efflux-pump was studied by using the efflux-pump inhibitor carbonyl cyanide-m-chlorophenylhydrazone (CCCP). Results: Resistance to ciprofloxacin in Shigella species increased from 0% in 2004 to 44% in 2010 and S. flexneri was the predominant species. Of Shigella spp, ciprofloxacin resistant (CipR) strains were mostly found among S. flexneri (8.3%), followed by S. sonnei (1.5%). Within S. flexneri (n = 2181), 14.5% were resistance to ciprofloxacin of which serotype 2a was predominant (96%). MIC of ciprofloxacin, norfloxacin, and ofloxacin were 6–32 mg/L, 8–32 mg/L, and 8–24 mg/L, respectively in S. flexneri 2a isolates. Sequencing of QRDR genes of resistant isolates showed double mutations in gyrA gene (Ser83Leu, Asp87Asn/Gly) and single mutation in parC gene (Ser80Ile). A difference in amino acid substitution at position 87 was found between strains isolated in Bangladesh (Asp87Asn) and China (Asp87Gly) except for one. A novel mutation at position 211 (His→Tyr) in gyrA gene was detected only in the Bangladeshi strains. Susceptibility to ciprofloxacin was increased by the presence of CCCP indicating the involvement of energy dependent active efflux pumps. A single PFGE type was found in isolates from Bangladesh and China suggesting their genetic relatedness. Conclusions: Emergence of fluoroquinolone resistance in Shigella undermines a major challenge in current treatment strategies which needs to be followed up by using empirical therapeutic strategies. [ABSTRACT FROM AUTHOR]

In this study, mechanisms of plasmid-mediated sulfamethoxazole resistances in the clinical strains of multi-drug resistant (MDR) Shigella flexneri 2a were elucidated for the first time in Bangladesh. From 2006 to 2011, a total of 200 S. flexneri 2a strains were randomly selected from the stock of the Enteric and Food Microbiology Laboratory of icddr,b. Antimicrobial susceptibility of the strains showed 73%, 98%, 93%, 58%, 98%, 64% and 4% resistance to trimethoprim-sulfamethoxazole, nalidixic acid, ampicillin, erythromycin, tetracycline, ciprofloxacin and ceftriaxone respectively. Plasmid profiling revealed heterogeneous patterns and interestingly, all the trimethoprim-sulfamethoxazole resistant (SXTR) strains yielded a distinct 4.3 MDa plasmid compared to that of the trimethoprim-sulfamethoxazole susceptible (SXTS) strains. Curing of this 4.3 MDa plasmid resulted in the susceptibility to sulfamethoxazole alone suggesting the involvement of this plasmid in the resistance of sulfamethoxazole. Moreover, PCR analysis showed the presence of sul2 gene in SXTR strains which is absent in SXTS strains as well as in the 4.3 MDa plasmid-cured derivatives, confirming the involvement of sul2 in the resistance of sulfamethoxazole. Furthermore, pulsed-field gel electrophoresis (PFGE) analysis revealed that both the SXTR and SXTS strains were clonal. This study will significantly contributes to the knowledge on acquired drug resistance of the mostly prevalent S. flexneri 2a and further warrants continuous monitoring of the prevalence and correlation of this resistance determinants amongst the clinical isolates of Shigella and other enteric pathogens around the world to provide effective clinical management of the disease. [ABSTRACT FROM AUTHOR]

The study was designed to develop a low calorie and shelf‐life extended fruit yogurt mixed with wood apple powder. The fruit powder was supplemented at different levels in order to recommend an appropriate formulation. From the proximate analyses, it was found that the major components such as protein, fat, and moisture contents varied slightly in the yogurt samples. However, the plain yogurt was found to contain a higher amount of carbohydrate (18.83%) than that of wood apple powder yogurt (12.72%). From the organoleptic responses, the yogurt sample containing 6% fruit powder and 20% table sugar secured the highest score in color, texture, and taste. A fruit mixed yogurt sample was prepared with as less as 30% low calorie content than the plain one. The fruit powder yogurts were found to have 5 to 6 days more shelf‐life than that of the plain yogurt at refrigeration temperature (7°C). Practical applications: The food items with low sugar content have proved to be very helpful in maintaining the soundness of health. The current study shows that only an appropriate formulation can provide healthier food, ensuring acceptable qualities. The study demonstrates how the natural fruit sugar and fruit acid contribute in preparing low calorie as well as shelf‐life extended yogurt. The laboratory investigations have suggested the appropriate formulation of wood apple yogurt with pros and cons regarding nutritive and organoleptic qualities. The findings of the study are highly expected to be helpful for the bakery and confectionary products manufacturers to prepare the yogurts with low calorie content and naturally enhanced shelf‐life. [ABSTRACT FROM AUTHOR]