Type Culture Research Articles

Ultrastructural characterization of hippocampal inhibitory synapses under resting and stimulated conditions.

The present study uses electron microscopy to document ultrastructural characteristics of hippocampal GABAergic inhibitory synapses under resting and stimulated conditions in three experimental systems. Synaptic profiles were sampled from stratum pyramidale and radiatum of the CA1 region from (1) perfusion fixed mouse brains, (2) immersion fixed rat organotypic slice cultures, and from (3) rat dissociated hippocampal cultures of mixed cell types. Synapses were stimulated in the brain by a 5min delay in perfusion fixation to trigger an ischemia-like excitatory condition, and by treating the two culture systems with 90 mM high K+ for 2-3min to depolarize the neurons. Upon such stimulation conditions, the presynaptic terminals of the inhibitory synapses exhibited similar structural changes to those seen in glutamatergic excitatory synapses, with depletion of synaptic vesicles, increase of clathrin-coated vesicles and appearance of synaptic spinules. However, in contrast to excitatory synapses, no structural differences were detected in the postsynaptic compartment of the inhibitory synapses upon stimulation. There were no changes in the appearance of material associated with the postsynaptic membrane or the length and curvature of the membrane. Also no change was detected in the labeling density of gephyrin, a GABAergic synaptic marker, lining the postsynaptic membrane. Furthermore, virtually all inhibitory synaptic clefts remained rigidly apposed, unlike in the case of excitatory synapses where ~ 20-30% of cleft edges were open upon stimulation, presumably to facilitate the clearance of neurotransmitters from the cleft. The fact that no open clefts were induced in inhibitory synapses upon stimulation suggests that inhibitory input may not need to be toned down under these conditions. On the other hand, similar to excitatory synapse, EGTA (a calcium chelator) induced open clefts in ~ 18% of inhibitory synaptic cleft edges, presumably disrupting similar calcium-dependent trans-synaptic bridges in both types of synapses.

Cultivation of Aurantiochytrium sp. Using Pickle Seasoning Liquid Waste and Application of the Raw Biomass for Fish Aquaculture

AbstractAurantiochytrium sp. strain L3W is a heterotrophic microorganism that produces docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), which are essential for the growth of marine fish. In this study, pickle seasoning liquid waste was used for culturing strain L3W and the raw biomass of strain L3W was then fed to red sea bream fingerlings (Pagus major) to confirm its usability as a source of DHA and EPA. The strain L3W was cultured on the three pickle seasoning liquid waste samples, Akimurasaki, Hiroshimana, Shisohijiki, diluted with sand-filtered seawater at initial pH values of 4 and 7 under unsterile conditions. The growth of strain L3W was highest at 3.71 g/L on Hiroshimana at an initial pH of 7 with DHA and EPA production at 71.4 and 6.8 mg/g-biomass, respectively. Preparing the raw biomass of strain L3W by the 200 L-scale cultivation using the American Type Culture Collection 790By+ medium because of the DHA and EPA contents close to that produced in the Hiroshimana medium with an initial pH of 7, the raw biomass was spiked to the diet at 3%, 5%, and 10%. The final DHA and EPA contents in the whole body were increased by 4.26 and 3.03 times, respectively, by adding the strain L3W biomass at 10%. These results confirmed the feasibility of a carbon cycling scenario in which the strain L3W is cultivated using liquid food waste with the resultant biomass is utilized as a source of DHA and EPA for fish aquaculture. Graphical Abstract

Complete genome sequence of Sphingobacterium thalpophilum BAA-1094.

We report the complete genome of Sphingobacterium thalpophilum BAA-1094 obtained from the American Type Culture Collection. The genome has one circular chromosome (5.7 Mbp). The rather low average nucleotide identity of 76.5% to the type strain Sphingobacterium thalpophilum DSM 11723 suggests the need for reclassification.

High Mortality Rates and Antimicrobial Resistance in Children with Invasive Bacterial Infections in a Tertiary Care Facility in West Africa

Abstract Background Childhood data on infection mortality and antimicrobial resistance in Sub-Saharan Africa are emerging but remain limited. The University of Maryland Center for Vaccine Development (CVD) and CVD-Mali have been conducting a prospective observational study at L’Hopital Gabriel Toure (HGT), a referral hospital in Bamako, Mali, since 2002 to identify causes of invasive bacterial infections (IBI) in children. These data have been used to initiate vaccination programs to decrease the burden of Haemophilus influenzae type b and Streptococcus pneumoniae. Additional demographic and antimicrobial susceptibility data have been collected since 2021, allowing for more detailed resistance and mortality analyses. Methods All children presenting to HGT are screened. Children ages 0-15 years with temperature of at least 39 degrees Celsius and/or suspicion of IBI are eligible. Eligible children have blood cultures collected with additional site-specific cultures when focal infection is suspected. Cultures are performed using the Bactec system; when positive, Gram staining, sub-culturing, and antimicrobial susceptibility testing (by disk diffusion using standard methods) are performed. Organisms thought to be contaminants are excluded. American type culture collection strains are utilized for quality control. Results Between 1/14/2021 and 9/14/2023, 28703 patients were screened. 2156 patients were included. 212 patients (10%) had 235 pathogens isolated. Gram positive organisms were most frequently isolated (N=112), followed by Gram negative Enterobacterales (N=81), non-Enterobacterales Gram negatives (N=36), and yeast (N=6). Staphylococcus aureus was the most frequently isolated Gram positive organism (59 bloodstream isolates, 2 joint isolates, 2 subcutaneous isolates) followed by S.pneumoniae (10 bloodstream isolates, 17 cerebrospinal fluid isolates, 1 joint isolate) and Enterococcus species (16 bloodstream isolates). Klebsiella pneumoniae (28 bloodstream isolates, 2 cerebrospinal fluid isolates) and Escherichia coli (18 bloodstream isolates, 3 cerebrospinal fluid isolates, 2 subcutaneous isolates) were the most frequently isolated Gram negative Enterobacterales, with Pseudomonas species most common among non-Enterobacterales Gram negative organisms (16 bloodstream isolates, 1 cerebrospinal fluid isolate). Mortality rates in both culture-positive and culture-negative patients were high (52% and 28%, respectively), with greater than 50% of deaths occurring within 48 hours in both groups. 88% of Gram negative Enterobacterales were ceftriaxone resistant or intermediate, with 56% ciprofloxacin resistant or intermediate; 55% of isolates were resistant or intermediate to both. 71% of isolates were gentamicin resistant. Imipenem resistance remained low, with 91% of isolates retaining sensitivity. 24% of S. aureus isolates were oxacillin resistant and 34% of isolates were resistant or intermediate to trimethoprim/sulfamethoxazole. 100% of S. aureus isolates tested (14/60) retained sensitivity to clindamycin. Only 54% of pneumococcal isolates had positive oxacillin screens, implying frequent penicillin resistance, and only 60% of enterococcal species were ampicillin sensitive, although 92% retained vancomycin sensitivity. Conclusion We found higher mortality rates in our study than in similar studies; the contribution of issues such as limited access to advanced supportive care and delay in presentation is an area for further study. The high frequency of multidrug-resistant organisms is alarming, suggesting an urgent need to investigate underlying contributing factors, increase access to affordable broad-spectrum antibiotics, and create robust antimicrobial stewardship programs.

In vitro antimalarial susceptibility profile of Plasmodium falciparum isolates in the BEI Resources repository.

BEI Resources, a National Institute of Allergy and Infectious Diseases-funded program managed by the American Type Culture Collection, serves researchers worldwide through the provision of a centralized repository for the acquisition, production, characterization, preservation, storage, and distribution of standardized biological resources targeting National Institutes of Health priority pathogens including bacteria, viruses, pathogenic fungi, and parasitic protozoa. These reference materials are critical for the development of diagnostics, vaccines, and therapeutics and are available to qualified registered investigators and institutions worldwide. Bioresources within BEI include well-characterized malaria isolates as part of the Malaria Research and Reference Reagent Resource Center (MR4). These isolates are critical for screening antimalarial compounds, conducting drug resistance studies, and for resistance surveillance and management. In our efforts to enhance the characterization of MR4 P. falciparum isolates, we measured antimalarial susceptibility of >100 isolates against a panel of standard antimalarial compounds. Our results provide valuable information to assist current and prospective users of the BEI Resources repository in making data-driven requests of isolates to meet their research needs.

The Influence of Selected Titanium Alloy Micro-Texture Parameters on Bacterial Adhesion.

The colonization of microbes and the resulting formation of biofilms on dental implants are significant contributors to peri-implantitis and the failure of these implants. The aim of the research was to analyze the impact of density and depth of laser texturing of the Ti-6Al-7Nb alloy surface on the colonization of selected microorganisms and biofilm formation. Standard strains of Gram-negative and Gram-positive bacteria and yeasts from the American Type Culture Collection-ATCC-were used to demonstrate the ability to form single-species biofilms in vitro. The study evaluated three types of titanium samples with different texture density and depth. The colonization and biofilm formation abilities of the tested microorganisms were assessed. The obtained results were subjected to statistical analysis. Among the analyzed strains, L. rhamnosus showed the highest colonization of the tested surfaces. It was found that there is no relationship between the texture parameters and the number of colony-forming units (CFU/mL) for C. albicans, S. mutans, and L. rhamnosus. For the F. nucleatum strain, it was shown that the number of colony-forming bacteria is related to the texture density.

Религия и национализм

The article analyzes the similarities and differences of ethnic and religious identities, their mutual influence and cases of joint impact on society. The author explores the role of these identities in the formation of the nation state. He also examines the ethnic and religious factor in heterogeneous societies, which are characterized by the presence of a significant number of diverse cultural groups united under one government. Russia also belongs to such societies. According to the author, in the current conditions in Russia, ethnic and religious identities cannot be used as a factor in nation building. The Russian form of government is more consistent with the imperial type. States of this type see the meaning and justification of their existence in protecting their constituent peoples, maintaining peace and order among them, and ensuring their development. Therefore, policy in the sphere of managing cultural and religious diversity in Russia cannot be built upon the model inherent in nation states, because the very term “nation” and nation building practices are associated with the division of people into “us” and “them”, unification, and suppression of minorities. Nationalism in Russia has always been and remains a risky project, fraught with the disintegration of the country into several dozen ethno-national states with the prospect of a military clash between them. The research conducted by the author shows that nationalism is most often not associated with religion, acting as secular worldview. In turn, religious traditions do not need national consolidation. The experience of the existence of the church in states and cultures of different types indicates that no ethnic culture or identity of a nation has enduring significance from a religious viewpoint.

First report of Rhizoctonia blight caused by Rhizoctonia solani on Chinese cabbage (Brassica rapa subsp. chinensis) in India.

During January and February 2021, foliar blight symptoms were observed on the leaves of Chinese cabbage (Pak choi) at Lembucherra research farm, College of Agriculture, Tripura, India. The incidence of disease symptoms ranged from 5 to 10% of the plants observed in the field. The symptomatic leaves showed grayish colored water-soaked lesions with an irreguar shape and size. A total of 10 symptomatic leaves (1 leaf per plant) from Chinese cabbage infected plant were sampled, surface decontaminated with 1% NaOCl, washed twice in sterile water, plated on 2% water agar, and incubated at 25 ± 2°C. Hyphal tips from mycelium of 7-day old culture (2 isolates from two different plants) with right-angled branching were transferred to potato dextrose agar (PDA) media (SRL, India). Cream or light brown hyphae that branched at right angles, with septa near the point of the origin of hyphae, and a slight constriction at the base of the branch) were visible under a microscope. Olive-brown sclerotia were observed after 5 days of incubation. Multiple nuclei per cell were visible after staining with 4', 6-diamidino-2-phenylindole (Estandarte et al. 2016). Based on morphological characteristics (Parmeter et al. 1970) the isolates TP36 and TP37 were identified as Rhizoctonia solani. The internal transcribed spacer (ITS) region and glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) were amplified with ITS1& ITS4 (White et al. 1990) and (GAPDH F-5'- CAAGGAGAACCCAGGTGTTAAG-3' and GAPDH R- 5'-GGCGTCGAAGATAGAAGAGTGT-3') respectively for both isolates and sequenced (accession #. PP458158, PP458159, PP425343, PP425344). BLASTn analysis showed 99.26%( 668/673 nt) to 99.46% (659/664 nt) identity with R. solani sequences (GenBank MG397062.1 and KX674524.1) for ITS and 98.42% (552/562 nt) to 100% 540/540 nt)identity with R. solani sequences (GenBank HQ425709.1 and CP102644.1) for GAPDH. Isolates TP36 and TP37 were deposited in the Indian Type Culture Collection (ITCC), New Delhi as R. solani (nos. 9154 and 9319, respectively). Both isolates were amplified using (anastomosis group) AG1 subgroup specific primers (Matsumoto 2002; Prashantha et al. 2021) to identify their AG. The presence of a 265 bp amplicon for both isolates suggested that they belong to AG1-IA. A multilocus analysis of R. solani isolates from different host plants with concatenated sequences ITS and GAPH showed that TP36 and TP37 are closely related to rice isolate RS107. A pathogenicity test on five plants per treatment was conducted and repeated twice on one month old Chinese cabbage plants (hybrid, TOKITA, India) grown under glasshouse conditions in a sterilized mixture of soil and sand (3:1) at 27-28oC during January 2024 at ICAR-IARI, New Delhi. R. solani isolates TP36 and TP37 were grown on PDA and plants were inoculated by placing single sclerotia of 10-day old colony on different plant parts and covering it with moist cotton. After 7 day, typical lesions of R. solani infection were visible. No symptoms were observed on the control plants. The fungus was reisolated from the inoculated plants and identified as R. solani based on morphology. R. solani has previously been reported to cause disease on some members of Brassicaceae in different countries (Budge et al. 2009; Hua et al. 2014). Based on literature available this is the first report of R. solani infecting Chinese cabbage in India.

Characterization of Electrochemically Active Bacteria Utilizing Redox Response in Microbial Electrolysis Cell

This study focuses on micro-electrochemical screening to select microbial strains capable of directly transfer electrons to the working electrode dependent on specific enzymatic machinery. The main objective of this work is to select and identify promising strains for allow the bioelectrolysis production of hydrogen. To achieve this goal, microbial composites (artificial biofilms), have been developed using Escherichia coli CGE1 from LCPME. CNRS Fransh, Pseudomonas putrifisciens (CIP 69.13) (CIP, Collection Institut Pasteur, Fransh). Shewanella oneidensis MR-1 (ATCC 700550), and Thiobacillus denitrificans, from (ATCC, American Type Culture Collection), each one enclosed in a matrix carbon nanotubes and protamine matrix, forming an artificial biofilm on buckypaper. Cyclic Voltammetry (CV) measurements were performed over a potential range of +0.4 to -0.7V at 5mV/s under 30°C, using a saturated KCl Ag/AgCl reference electrode and a stainless-steel grid counter electrode. For E. coli and P. putrifisciens, the measurement focused on the oxidation of 20mM glucose, while the former bacteria were growth with and without O2. For S. oneidensis and T. denitrificans the focus was on the reduction of fumarate and 20 mM of NaNOH3+, respectively. As results, E. coli and P. putrifisciens species show no notable electrochemical activity, with no signal of glucose oxidation, due to the absence of type C cytochromes in the cytoplasmic membrane, unlike S. oneidensis and T. denitrificans, that demonstrate a direct electron transfer.

The Origin of the Problem: Characterization of Paraguayan Septoria steviae, Causal Agent of Septoria Leaf Spot of Stevia, Based on Multilocus Sequence Analysis.

Septoria leaf spot is a significant disease affecting cultivated stevia, potentially reducing yields by > 50%. The disease is caused by Septoria steviae, first identified in 1978 in Japan as a new pathogen of stevia. Understanding the origin of S. steviae could clarify how it spread to new production areas. To investigate this, 12 isolates of Septoria sp. were obtained from stevia's native range in the Amambay forests and field plantings in Paraguay from 2018 to 2020. These isolates underwent colony morphology and molecular characterization of Actin, β-Tubulin, Calmodulin, ITS, LSU, RPB2, and TEF1α loci. GenBank sequences from S. steviae isolates collected in France, Japan, and the United States were included. Multilocus sequence phylogenetic analysis generated a maximum likelihood (ML) tree. The morphological characteristics of Paraguayan isolates were similar to those of previously reported S. steviae type cultures from Japan. The ML analysis showed that Paraguayan isolates formed a monophyletic group with S. steviae isolates from France, Japan, and the United States. During blotter tests, pycnidia and cirri of S. steviae were observed on multiple stevia seed surfaces from different sources. Further characterization confirmed viable pathogenic conidia of S. steviae. This observation suggests that S. steviae could be associated with stevia seed, possibly spreading from the center of origin to other countries. This research is the first to genetically characterize S. steviae from Paraguay and propose its potential spread mechanism from the center of origin to the rest of the world.

Comparative Study on Quorum Modulatory Effect of Selected Medicinal Plants on Chromobacterium violaceum ATCC 12472 (MTCC 2656)

Among the main global health concern is the rampant rise in antibiotic resistant bacteria. One of the appealing and promising strategies to combat this menace is to target the adaptive mechanism called quorum sensing (QS) used by bacteria to survive. Exploratory research on anti-QS compounds derived from natural products has been a promising area. The present study investigated methanolic extracts from 26 plants to compare their anti-QS activity using the QS biosensor strain Chromobacterium violaceum American Type Culture Collection (ATCC 12472) (Microbial Type Culture Collection MTCC2656). QS-mediated violacein pigment inhibition was carried out using agar well diffusion method with concentrations ranging from 10 mg/ml to 100 mg/ml. Leaf extracts of Mangifera indica and Pimenta dioica and peel extract of Punica granatum were the only three plants found to exhibit violacein inhibitory potential till 10 mg/ml. The result of the minimum inhibitory concentration (MIC) showed 1.6 mg/ml for M. indica and P. dioica and 6.25 mg/ml for P. granatum. Further, violacein inhibitory properties of these extracts at and below MIC were evaluated by well diffusion assay (qualitative) and by flask incubation assay (quantitative). The zone of inhibition (well diffusion assay) was found to be 14.51 ± 0.63 mm to 10.37 ± 0.68mm for M. indica, 15.23 ± 0.57 mm to 9.62 ± 1.29 mm for P. granatum and 17.01 ± 0.1 mm to 13.14 ± 0.18 mm for P. dioica. The inhibitory effect of the plant extracts via quantitative assay on violacein ranged from 83-49%, 89-81%, and 89-49% for M. indica, P. granatum, and P. dioica respectively. Our findings suggested the potential of M. indica, P. granatum, and P. dioica methanolic extracts as a source of effective inhibitors of QS-mediated violacein production.

Synthesis of Fe3O4@SiO2–branched polyethylenimine nanospheres for removal of Cr(VI) and anionic dyes

There is a need to design and prepare new sorbents for the effective elimination of toxic metal ions and textile dyes from the environment. In the presented study, the preparation of magnetic functionalized silica composite nanospheres for the removal of inorganic and organic pollutants from aqueous solutions is reported. Firstly, the surface of silica coated magnetic nanospheres was functionalized with epoxy groups via a reaction of 3-glycidoxypropyltriethoxysilane. Then, the branched polyethylenimine (BPEI) was attached to generate positive functional charged groups on the on the Fe3O4@SiO2. The performance of the Fe3O4@SiO2-BPEI nanospheres for the removal of Cr(VI) and Congo Red (CR) and Trypan Blue (TB) dyes was studied under various experimental conditions. The optimum pHs values for adsorption of Cr(VI) ions, CR and TB dyes on the Fe3O4@SiO2-BPEI nanospheres were found to be at pH 2.0, and at pH 4.0, respectively. The maximum adsorption capacities of the Fe3O4-@SiO2-BPEI nanospheres for Cr(VI), CR, and TB were obtained as 356.7, 395.3, and 561.8 mg/g within 90 min equilibrium time, an initial concentration of 500 mg/L, and at 25 ºC, respectively. The adsorption of the tested pollutants were well described by the Langmuir isotherm model and second-order kinetic model. After ten reuse cycles, the adsorption capacity of Fe3O4-@SiO2-BPEI nanospheres was not significantly changed for the tested pollutants. The adsorption mechanism of Cr(VI), CR, and TB by the Fe3O4-@SiO2-BPEI nanospheres was evaluated to be related to ion exchange and ion exchange and/or hydrogen bonding interactions, respectively. The results also suggested that the presented adsorbent may serve as a multi-functional adsorbent for the remediation of industrial effluents. In addition, compared to the unmodified Fe3O4@SiO2, the Fe3O4@SiO2-BPEI nanospheres have antibacterial activities against Escherichia coli (American Type Culture Collection (ATCC) 12435), Pseudomonas aeruginosa (ATCC 10145), Listeria monocytogenes (ATCC 7644), and Staphylococcus aureus (ATCC 43300). Based on the obtained results from this work, the branched polyethylene functionalized magnetic silica nanospheres with antibacterial properties can be used as a potential adsorbent for the removal of metal ions and anionic dyes from wastewater.

Isolation and identification of Cladosporium tenuissimum causing leaf blight on garden pea (Pisum sativum L.) in Telangana from southern India

Garden pea (Pisum sativum L.), is an important legume grown worldwide as a source of proteins and carbohydrates. Since pea crop is cultivated globally in cool temperate climatic conditions, increasing temperatures accompanied by high humidity favors fungal infections. A fungal infection causing leaf blight symptoms was frequently observed on the leaves of pea plants grown in greenhouse and field conditions. The blight symptoms showed a water-soaked and wilting appearance with curly leaf edges accompanied by greyish mycelial growth and chlorotic lesions. The disease incidence in field conditions ranged between 5 and 10 %, whereas in the controlled greenhouse conditions, it ranged from 40 to 45%. Two isolates of the fungus, Gp03 and Gp04 were isolated from Arkel pea leaves and purified from a single conidium each. According to Koch's postulates, the pathogenicity tests with these isolates using detached leaf assay and whole plant leaf assays confirmed the pathogen. Morphological and molecular analysis of nucleotide sequences of the D1-D2 region, translation elongation factor-1α (TEF) and actin (ACT) genes confirmed these isolates as members of the genus, Cladosporium. The phylogenetic relatedness of these isolates with other members of the Cladosporium genus revealed them as fungal strains of Cladosporium tenuissimum. Further, these isolates were also confirmed as C. tenuissimum by Microbial Type Culture Collection and Gene Bank (MTCC), Chandigarh in India, and provided the accession nos. MTCC 13581 and MTCC 13582 to the C. tenuissimum strains, Gp03 and Gp04 respectively. This is the first report of Cladosporium tenuissimum infecting garden pea in Telangana, India.

Anti-biofilm activity of 445nm and 970nm diode laser on mixed species colonies of- aggregatibacter actinomycetemcomitans and porphyromonas gingivalis cultured on titanium discs -an in vitro study.

To assess and compare the anti-microbial efficacy of 445nm and 970nm diode laser on mixed species biofilm of Aggregatibacter actinomycetemcomitans [A.a] and Porphyromonas gingivalis [P.g] cultured on machined pure titanium discs. A total of 65 machined pure titanium discs with no surface modifications with a 10-mm diameter and a 2-mm height were sterilized by autoclaving at 121°C for 15min and incubated with the commercially available bacterial strains ATCC(American Type Culture Collection- P.g 33277 and A.a 29522)mixture of Aggregatibacter actinomycetemcomitans(A.a) and Porphyromonas gingivalis(P.g).After a 2-week incubation period with the mixture of bacteria to develop a mixed species biofilm, the discs were divided into three groups: (1) no treatment (control), (2) 445nm laser (test), (3) 970nm laser (test). For each laser wavelength (445 and 970nm), the discs were exposed to 1.0W and 2.0W in continuous wave mode for the times points of 15, 30, and 60s. The antimicrobial efficacy was assessed by qPCR. A significant reduction in the levels of both species of bacteria was observed between control and the laser intervention groups. A higher efficacy for the 445nm diode laser against Porphyromonas gingivalis and a similar efficacy against Aggregatibacter actinomycetemcomitans was observed as compared to the 970nm group. 445nm wavelength represents a potential and effective laser wavelength which can be used for the management of peri-implant infection. The present study findings also need to be further validated through clinical interventional trials.

DETERMINATION OF THE CIRCADIAN OSCILLATION PATTERN OF UNFOLDED PROTEIN RESPONSE SIGNALING COMPONENTS IN HUMAN EMBRYONIC KIDNEY HEK293 CELLS

Objective: The circadian rhythm is one of the primary regulatory systems with near 24-hour oscillations. It has a crucial role in regulating physiological conditions in the human body, including body temperature and the secretion of hormones. Numerous disorders, such as cancer and diabetes, have been linked to disruptions of the cellular circadian rhythm. Herein, we aimed to investigate the relationship between the circadian rhythm and unfolded protein response (UPR) signaling, which is one of the important physiological mechanisms in mammalian cells and has recently been associated with drug resistance, invasion and metastasis in cancer. Material and Method: Human embryonic kidney cell line HEK293 was provided from the American Type Culture Collection and propagated in DMEM containing 10% FBS and growth ingredients. For in vitro circadian synchronization, cells were exposed to 50% and then the oscillation pattern of gene and protein expression of UPR-related target genes was analyzed by agarose gel electrophoresis and immunoblotting, respectively. The oscillation pattern was commented on through curve-fitting analysis. Result and Discussion: Our findings demonstrated that UPR components, including IRE1α, XBP-1s, eIF2α, phospho(Ser51)-eIF2α, PERK, ATF4, GADD34 and ATF6, tightly exhibit oscillation patterns under a circadian rhythm on a 48-hour time scale like the PER1 gene that is a core component of the circadian rhythm. Moreover, endoplasmic reticulum (ER) stress genes, BiP/GRP78 and CHOP, were similar to UPR components under the circadian rhythm. Additionally, we found the activation of UPR signaling harmoniously modulated with the circadian rhythm. Present data indicated that the expression level of UPR components exhibited strict oscillation under the circadian rhythm. Our findings may guide experimental studies of new-generation UPR-targeted drugs to be developed to treat various pathologies in accordance with the circadian rhythm.

First report of Phytopythium vexans causing gummosis and root rot of Khasi Mandarin (Citrus reticulata Blanco) in north eastern states of India.

Khasi mandarin (Citrus reticulata Blanco) is the most economically important crop among the citrus growing region in the north-eastern India (Singh et al. 2016). An extensive survey was conducted to identify the causal agent of citrus root rot and gummosis in north eastern states (Meghalaya, Tripura, Manipur, Arunachal Pradesh, Sikkim, Nagaland and Assam) of India during October 2021-23. The gummosis disease incidence ranged from 5 to 95 % in 10 to 25 years old Khasi mandarin plants showing relatively more chronic symptoms on mature trees. Yellowing and dropping of leaves, twigs die back, gum oozing from infected bark and loss of feeder roots were the typical symptoms of the disease. Infected bark tissue and young lemon leaf baits in rhizosphere soil were plated on corn meal agar medium supplemented with pimaricin (10 µg/ml), ampicillin (250 µg/ml), rifamycin (10 µg/ml) and 300µg/ml carbendazim and incubated at 26℃. Fifty isolates were purified and maintained on Carrot agar medium. These isolates showed similar cultural and morphological characteristics. Two representative isolates from Arunachal Pradesh (AP21 and AP26) were selected for further experiments and deposited to Indian Type culture collection (ITCC), New Delhi with accession no. 9156 and 9157 respectively. The colonies were fast growing, showing rosette pattern along with whitish blooming mycelium appearance with no visual sporulation at the surface. The hyphae were coenocytic with initially right-angled branching. Sporangia were globose or sub globose and papillated. Oogonia were smooth and globose (16.29-21.09 µm) in diameter. Antheridia were irregular, cylindrical and broadly attached to oogonia. Empty sporangia were also observed. Multilocus phylogenetic analysis using internal transcribed spacer region (Das et al. 2011), β tubulin (Blair et al. 2008) and Cytochrome oxidase II gene (Noireung et al. 2020) showed that these isolates formed a stable clade with Phytopythium vexans (CBS119.80) sequence retrieved from NCBI database. BLAST analysis showed that ITS sequence of AP21 (OQ372986) and AP26 (OQ381083) had >99 % identity with P. vexans isolate NS-3 (ON533631). Further, BLAST analysis of β tubulin (AP21 OQ446053, AP26 OR405377) and Cox II gene (AP21 OQ473414, AP26 OR552422) sequences showed that our Indian isolates showed >99 % similarity with P. vexans voucher strain CBS119.80. To fulfil Koch's postulates, Khasi mandarin (Citrus reticulata) seedlings were inoculated by adding 100 ml zoospore suspension of P. vexans (1x105 spores/ml) in sterilized soil (Thao et al. 2020). The experiment was carried out in triplicate. Yellowing of leaves and leaf drop were observed 7 days post inoculation while 30 days post inoculation, treated plants started showing symptoms of root rot, including mild root decay. No symptoms were observed in control treatment. The pathogen was reisolated from symptomatic roots and confirmed through colony and sporangium morphology. Recently, it was reported that P. vexans is associated with apple and pear decline in the Saiss plain of Morocco (Jabiri et al. 2021), root rot on mandarin in Thailand (Noireung et al. 2020) and on Durian in Vietnam (Thao et al. 2020). As per our knowledge, this is the first report of P. vexans causing root rot and gummosis in Khasi mandarin from north eastern states of India. This finding is significantly important for the development of a successful disease management strategy in India.

Antimicrobial Effect of Passiflora foetida Leaf Extract

Antimicrobial resistance (AMR) is a critical global health concern under the present clinical scenario due to the overuse and misuse of antibiotics in both medical and agricultural sectors. Passiflora foetida, commonly known as wild maracuja or stinking passion flower belongs to the Passifloraceae family and is native to tropical and subtropical regions of Asia, Africa, Australia, and America. In this study, we aimed to explore the potential antimicrobial efficacy of Passiflora foetida leaf extract against both MDR bacteria and American Type Culture Collection (ATCC) strains for possible combating microbial infections in the future. The MIC value of Passiflora foetida extract varied between 5-10 mg/ml in different ATCC and MDR bacteria studied in this experiment. Passiflora foetida is also well known in traditional medicine indicating a degree of safety and efficacy, although modern scientific validation is necessary to support its therapeutic applications. Future research should focus on identifying the specific bioactive agent responsible for its antimicrobial effects, optimizing extraction methods to maximize bioavailability, and evaluating its safety and efficacy in animal models and clinical trials. Understanding the antimicrobial properties of Passiflora foetida is crucial for developing effective therapeutic strategies against antibiotic-resistant infections. By harnessing the natural antimicrobial potential of this plant, researchers may uncover new treatment options to combat AMR and improve patient outcomes. The findings of this study indicated its possible use as a topical application at present till further study with refined extract may reveal its application in vivo in different diseases with MDR microorganisms.

Exploring prebiotic properties and its probiotic potential of new formulations of soy milk-derived beverages.

The food and beverage industry has shown a growing interest in plant-based beverages as alternatives to traditional milk consumption. Soy milk is derived from soy beans and contains proteins, isoflavones, soy bean oligosaccharides, and saponins, among other ingredients. Because of its high nutritive value and versatility, soy milk has gained a lot of attention as a functional food. The present work aims to explore the prebiotic properties and gastrointestinal tolerance potential of new formulations of soy milk-derived drinks to be fermented with riboflavin-producing probiotic Lactiplantibacillus plantarum MTCC (Microbial Type Culture Collection and Gene Bank) 25432, Lactiplantibacillus plantarum MTCC 25433, and Lactobacillus acidophilus NCIM (National Collection of Industrial Microorganisms) 2902 strains. The soy milk co-fermented beverage showed highest PAS (1.24 ± 0.02) followed by soy milk beverages fermented with L. plantarum MTCC 25433 (0.753 ± 0.0) when compared to the commercial prebiotic raffinose (1.29 ± 0.01). The findings of this study suggested that the soy milk beverages exhibited potent prebiotic activity, having the ability to support the growth of probiotics, and the potential to raise the content of several bioactive substances. The higher prebiotics activity score showed that the higher the growth rate of probiotics microorganism, the lower the growth of pathogen. For acidic tolerance, all fermented soy milk managed to meet the minimal requirement of 106 viable probiotic cells per milliliter at pH 2 (8.13, 8.26, 8.30, and 8.45 logs CFU/mL, respectively) and pH 3.5 (8.11, 8.07, 8.39, and 9.01 log CFU/mL, respectively). The survival rate of soy milk LAB isolates on bile for 3 h ranged from 84.64 to 89.60%. The study concluded that lactobacilli could thrive in gastrointestinal tract. The sensory evaluation scores for body and texture, color, flavor, and overall acceptability showed a significant difference (p < 0.05) between the fermented probiotic soy milk and control samples. Soy milk fermented with a combination of L. plantarum MTCC 25432 & MTCC 25433 demonstrated the highest acceptability with the least amount of beany flavor. The findings of the study suggest soy milk's potential in plant-based beverage market.

Immunogenicity and vaccine potential of clinical isolate Mycobacterium kansasii strain against Mycobacterium tuberculosis infection.

Mycobacterium kansasii is a bacterium included in non-tuberculous mycobacteria (NTM) that can cause lung disease. It shares a significant number of antigens with Mycobacterium tuberculosis (Mtb), suggesting that it has the potential to be used as a tuberculosis (TB) vaccine. Therefore, we subcutaneously vaccinated mice with reference strain, M. kansasii-ATCC12478 [M. kansasii-American Type Culture Collection (ATCC)], and clinically isolated strain, M. kansasii-SM-1 to evaluate potential as a TB vaccine by comparing with bacille Calmette-Guerin (BCG) vaccine. Ten weeks after vaccination, we evaluated immunogenicity of M. kansasii-ATCC and M. kansasii-SM-1, and M. kansasii-SM-1 immunization induces potent Mtb antigen-specific IFN-γ-producing CD4+ T cells than M. kansasii-ATCC. Upon Mtb infection, M. kansasii-SM-1 provided better protection than M. kansasii-ATCC, which was comparable to the efficacy of BCG. These results showed that the clinical strain M. kansasii-SM-1, which exhibits an enhanced Mtb antigen-specific Th1 response, shows greater vaccine efficacy compared to M. kansasii-ATCC. In this study, we demonstrated that vaccine efficacy can vary depending on the strain of M. kansasii and that its efficacy can be comparable to BCG. This suggests that M. kansasii has the potential to be a live TB vaccine candidate.IMPORTANCEMycobacterium kansasii, a non-tuberculous mycobacteria (NTM) species causing lung disease, shares key antigens with Mycobacterium tuberculosis (Mtb), indicating its potential for TB vaccine development. Subcutaneous vaccination of mice with M. kansasii strains reference strain M. kansasii-ATCC12478 [(M. kansasii-American Type Culture Collection (ATCC)] and clinically isolated strain M. kansasii-SM-1 revealed differences in immunogenicity. M. kansasii-SM-1 induced a robust Mtb antigen-specific IFN-γ-producing CD4+ T cell response compared to M. kansasii-ATCC. Additionally, M. kansasii-SM-1 conferred better protection against Mtb infection than M. kansasii-ATCC, which is comparable to bacille Calmette-Guerin (BCG). These findings underscore the variable vaccine efficacy among M. kansasii strains, with M. kansasii-SM-1 exhibiting promising potential as a live TB vaccine candidate, suggesting its comparative effectiveness to BCG.

A New Guanidine-Core Small-Molecule Compound as a Potential Antimicrobial Agent against Resistant Bacterial Strains.

The guanidine core has been one of the most studied functional groups in medicinal chemistry, and guanylation reactions are powerful tools for synthesizing this kind of compound. In this study, a series of five guanidine-core small molecules were obtained through guanylation reactions. These compounds were then evaluated against three different strains of Escherichia coli, one collection strain from the American Type Culture Collection (ATCC) of E. coli ATCC 35218, and two clinical extended-spectrum beta-lactamase (ESBL)-producing E. coli isolates (ESBL1 and ESBL2). Moreover, three different strains of Pseudomonas aeruginosa were studied, one collection strain of P. aeruginosa ATCC 27853, and two clinical multidrug-resistant isolates (PA24 and PA35). Among Gram-positive strains, three different strains of Staphylococcus aureus, one collection strain of S. aureus ATCC 29213, and two clinical methicillin-resistant S. aureus (MRSA1 and MRSA2) were evaluated. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) experiments were reported, and the drop plate (DP) method was used to determine the number of viable suspended bacteria in a known beaker volume. The results from this assessment suggest that guanidine-core small molecules hold promise as therapeutic alternatives for treating infections caused by clinical Gram-negative and Gram-positive bacteria, highlighting the need for further studies to explore their potential. The results from this assessment suggest that the chemical structure of CAPP4 might serve as the basis for designing more active guanidine-based antimicrobial compounds, highlighting the need for further studies to explore their potential.