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1.
Arch Virol ; 169(7): 145, 2024 Jun 12.
Artículo en Inglés | MEDLINE | ID: mdl-38864875

RESUMEN

Since 2020, African swine fever (ASF) has affected all pig breeds in Northeast India except Doom pigs, a unique indigenous breed from Assam and the closest relatives of Indian wild pigs. ASF outbreaks result in significant economic losses for pig farmers in the region. Based on sequencing and phylogenetic analysis of the B646L (p72) gene, it has been determined that ASFV genotype II is responsible for outbreaks in this region. Recent studies have shown that MYD88, LDHB, and IFIT1, which are important genes of the immune system, are involved in the pathogenesis of ASFV. The differential expression patterns of these genes in surviving ASFV-infected and healthy Doom breed pigs were compared to healthy controls at different stages of infection. The ability of Doom pigs to withstand common pig diseases, along with their genetic resemblance to wild pigs, make them ideal candidates for studying tolerance to ASFV infection. In the present study, we investigated the natural resistance to ASF in Doom pigs from an endemic area in Northeast India. The results of this study provide important molecular insights into the regulation of ASFV tolerance genes.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Brotes de Enfermedades , Filogenia , Animales , Fiebre Porcina Africana/virología , Fiebre Porcina Africana/epidemiología , Fiebre Porcina Africana/inmunología , Virus de la Fiebre Porcina Africana/genética , Virus de la Fiebre Porcina Africana/inmunología , India/epidemiología , Porcinos , Brotes de Enfermedades/veterinaria , Genotipo , Factor 88 de Diferenciación Mieloide/genética , Resistencia a la Enfermedad/genética
2.
Arch Virol ; 169(3): 54, 2024 Feb 21.
Artículo en Inglés | MEDLINE | ID: mdl-38381218

RESUMEN

African swine fever (ASF) has emerged as a threat to swine production worldwide. Evasion of host immunity by ASF virus (ASFV) is well understood. However, the role of ASFV in triggering oncogenesis is still unclear. In the present study, ASFV-infected kidney tissue samples were subjected to Illumina-based transcriptome analysis. A total of 2463 upregulated and 825 downregulated genes were differentially expressed (p < 0.05). A literature review revealed that the majority of the differentially expressed host genes were key molecules in signaling pathways involved in oncogenesis. Bioinformatic analysis indicated the activation of certain oncogenic KEGG pathways, including basal cell carcinoma, breast cancer, transcriptional deregulation in cancer, and hepatocellular carcinoma. Analysis of host-virus interactions revealed that the upregulated oncogenic RELA (p65 transcription factor) protein of Sus scrofa can interact with the A238L (hypothetical protein of unknown function) of ASFV. Differential expression of oncogenes was confirmed by qRT-PCR, using the H3 histone family 3A gene (H3F3A) as an internal control to confirm the RNA-Seq data. The levels of gene expression indicated by qRT-PCR matched closely to those determined through RNA-Seq. These findings open up new possibilities for investigation of the mechanisms underlying ASFV infection and offer insights into the dynamic interaction between viral infection and oncogenic processes. However, as these investigations were conducted on pigs that died from natural ASFV infection, the role of ASFV in oncogenesis still needs to be investigated in controlled experimental studies.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Neoplasias Hepáticas , Animales , Porcinos , Virus de la Fiebre Porcina Africana/genética , Transcriptoma , Fiebre Porcina Africana/genética , Oncogenes , Transformación Celular Neoplásica , Carcinogénesis/genética
3.
Mol Biol Rep ; 51(1): 195, 2024 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-38270707

RESUMEN

BACKGROUND: The angiogenic cytokine vascular endothelial growth factor A (VEGFA) also exerts non-angiogenic effects on endocrine functionality of porcine luteal cells critical for progesterone (P4) production. METHOD AND RESULTS: The expression dynamics of VEGFA-FLT/KDR system were investigated using RT-qPCR during luteal stages and VEGFA gene knock out (KO) porcine luteal cells were generated using CRISPR/Cas9 technology. The downstream effects of VEGFA ablation were studied using RT-qPCR, Annexin V, MTT, ELISA for P4 estimation and scratch wound assay. Bioinformatics analysis of RNA-Seq data of porcine mid-luteal stage was conducted for exploring protein-protein interaction network, KEGG pathways, transcription factors and kinase mapping for VEGFA-FLT/KDR interactomes. The VEGFA-FLT/KDR system expressed throughout the luteal stages with highest expression during mid- luteal stage. Cellular morphology, structure and oil-red-o staining for lipid droplets did not differ significantly between VEGFA KO and wild type cells, however, VEGFA KO significantly decreased (p < 0.05) viability and proliferation efficiency of edited cells on subsequent passages. Expression of apoptotic gene, CASP3 and hypoxia related gene, HIF1A were significantly (p < 0.05) upregulated in KO cells. The relative mRNA expression of VEGFA and steroidogenic genes STAR, CYP11A1 and HSD3B1 decreased significantly (p < 0.05) upon KO, which was further validated by the significant (p < 0.05) decrease in P4 output from KO cells. Bioinformatics analysis mapped VEGFA-FLT/KDR system to signalling pathways associated with steroidogenic cell functionality and survival, which complemented the findings of the study. CONCLUSION: The ablation of VEGFA gene resulted in decreased steroidogenic capability of luteal cells, which suggests that VEGFA exerts additional non-angiogenic regulatory effects in luteal cell functionality.


Asunto(s)
Sistemas CRISPR-Cas , Células Lúteas , Femenino , Porcinos , Animales , Sistemas CRISPR-Cas/genética , Edición Génica , Factor A de Crecimiento Endotelial Vascular/genética , Anexina A5
4.
J Food Sci Technol ; 61(8): 1516-1524, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38966784

RESUMEN

Escherichia coli and Staphylococcus aureus are the most important food borne pathogen transmitting from animal meat and meat products. Therefore, it is vital to design an accurate and specific diagnostic tool for identifying those food-borne pathogens in animal meat and meat products. In the current study, E. coli, methicillin-resistant and sensitive S. aureus (MRSA and MSSA) were simultaneously detected using a developed triplex PCR-based technique. To obtain an optimal reaction parameter, the multiplex assay was optimised by changing just one parameter while holding the others constant. Specificity of the assay was assessed using several porcine bacterial template DNA. The plasmid DNA was used to test the multiplex PCR assay's sensitivity and interference in spiked pork samples. E. coli, MRSA, and MSSA each have PCR amplified products with sizes of 335, 533, and 209 bp, respectively. The assay detects a minimum microbial load of 102 CFU/µl for all the three pathogens and can identify bacterial DNA as low as 10-2 ng/µl. The assay was validated employing 210 pork samples obtained from retail meat shops and slaughter houses, with MRSA, E. coli, and MSSA with the occurrence rate of 1.9%, 42.38%, and 18.1%, respectively. The rate of mixed bacterial contamination in pork meat samples examined with the developed method was 6.19%, 1.43%, 1.90%, and 1.43% for MSSA & E. coli, MRSA & E. coli, MSSA & MRSA, and E. coli, MSSA & MRSA, respectively. The developed multiplex PCR assay is quick and efficient, and it can distinguish between different bacterial pathogens in a single reaction tube.

5.
Microb Pathog ; 185: 106452, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37972743

RESUMEN

The present investigation focuses on examining the clinical, histopathological, and ultrastructural changes that occurred in pig, during an outbreak of African swine fever (ASF) in 2022 in Assam, India. The disease initially manifested as a per-acute case with high mortality but without any evident clinical signs. Subsequently, some animals exhibited an acute form of the disease characterized by high fever (104-106 °F), anorexia, vomiting, respiratory distress, and bleeding from the anal and nasal orifices. During acute African swine fever virus (ASFV) infections, elevated levels of pro-inflammatory IL-1α, IL-1ß, IL-6, TNF, CCL2, CCL5, and CXCL10 were detected in the palatine tonsil, lymph nodes, spleen, and kidney using qPCR assay. These molecular changes were associated with haemorrhages, edemas, and lymphoid depletion. Postmortem examinations revealed prominent features such as splenomegaly with haemorrhages, haemorrhagic lymphadenitis, severe petechial haemorrhage in the kidney, pneumonia in the lungs, and necrotic palatine tonsil. Histopathological analysis demonstrated lymphocyte depletion in lymphoid organs, multi-organ haemorrhages, and interstitial pneumonia in the lungs. Scanning electron microscopy (SEM) further confirmed lymphocyte depletion in lymphoid organs through lymphocyte apoptosis and kidney damage with distorted tubules due to red blood cell destruction. Transmission electron microscopy reaffirmed lymphocyte apoptosis by observing chromatin condensation and nucleus margination in lymphocytes of lymphoid organs. These findings provide comprehensive insights into the clinical, histopathological, and ultrastructural aspects of ASF outbreak in pigs. Understanding the pathological changes associated with ASF can contribute to improved diagnosis, prevention, and control measures for this highly contagious and economically devastating viral disease.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Porcinos , Animales , Fiebre Porcina Africana/epidemiología , Fiebre Porcina Africana/patología , Linfocitos , Brotes de Enfermedades , Hemorragia , Sus scrofa
6.
Phys Chem Chem Phys ; 25(20): 14374-14386, 2023 May 24.
Artículo en Inglés | MEDLINE | ID: mdl-37184034

RESUMEN

A Cu(II)-salen complex encapsulated in MWW-framework as an efficient chiral organocatalyst was developed for the synthesis of 3,4-dihydropyrimidin-2-(1H)-one (DHPMs) derivatives via an asymmetric pathway. In order to confirm its structural properties, single-crystal X-ray diffraction, powder XRD, BET, XPS, FE-SEM, EDX, UV-Vis, and FTIR spectra were used. Using computer-assisted DFT calculations, the Cu(II)-salen complex has been fine-tuned to fit into the pocket of the porous MWW support while keeping its chirality. This organocatalyst was shown to be a potent catalyst for the formation of the desired DHPMs product under short reaction times. Furthermore, this green protocol allows rapid and simple isolation of active MWW-trapped Cu(II)-salen scaffolds and its reusability in at least five consecutive runs without losing much of its activity.

7.
Environ Res ; 231(Pt 1): 115862, 2023 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-37146933

RESUMEN

Cancer is a major global public health concern that affects both industrialized and developing nations. Current cancer chemotherapeutic options are limited by side effects, but plant-derived alternatives and their derivatives offer the possibilities of enhanced treatment response and reduced side effects. A plethora of recently published articles have focused on treatments based on cannabinoids and cannabinoid analogs and reported that they positively affect healthy cell growth and reverse cancer-related abnormalities by targeting aberrant tumor microenvironments (TMEs), lowering tumorigenesis, preventing metastasis, and/or boosting the effectiveness of chemotherapy and radiotherapy. Furthermore, TME modulating systems are receiving much interest in the cancer immunotherapy field because it has been shown that TMEs have significant impacts on tumor progression, angiogenesis, invasion, migration, epithelial to mesenchymal transition, metastasis and development of drug resistance. Here, we have reviewed the effective role of cannabinoids, their analogs and cannabinoid nano formulations on the cellular components of TME (endothelial cells, pericytes, fibroblast and immune cells) and how efficiently it retards the progression of carcinogenesis is discussed. The article summarizes the existing research on the molecular mechanisms of cannabinoids regulation of the TME and finally highlights the human studies on cannabinoids' active interventional clinical trials. The conclusion outlines the need for future research involving clinical trials of cannabinoids to demonstrate their efficacy and activity as a treatment/prevention for various types of human malignancies.


Asunto(s)
Cannabinoides , Neoplasias , Humanos , Cannabinoides/farmacología , Células Endoteliales , Transición Epitelial-Mesenquimal , Neoplasias/tratamiento farmacológico , Microambiente Tumoral , Ensayos Clínicos como Asunto
8.
Biofouling ; 39(2): 171-188, 2023 02.
Artículo en Inglés | MEDLINE | ID: mdl-37057638

RESUMEN

In an era of antibiotic resistance where natural antibiotic substitutes are considered essential, the antimicrobial and antibiofilm activities of Citrus limon extract on strains of pathogenic Escherichia coli isolated from pork were evaluated. The strains which form biofilms were more resistant (MIC50 = 2.5 mgml-1) compared to non-biofilm forming strains (MIC50 = 1.25 mgml-1). Use of C. limon extract at 20 mgml-1 concentration has resulted in inhibition of biofilm formation by 53.96%. Cyclobarbital, 5, 8-dimethoxycumarin, orotic acid and 3-methylsalicylhydrazide were the major phytochemicals in C. limon extract with highest docking affinities against the biofilm associated proteins in E. coli. The results of simulation studies have clearly illustrated the energy stability of the protein-ligand complexes. Absorption, distribution, metabolism, excretion and toxicity (ADMET) profiles revealed that the phytochemicals in C. limon could be used in the drug design studies to preferentially target the specific receptors to combat biofilms associated with E. coli.


Asunto(s)
Citrus , Escherichia coli , Biopelículas , Proteínas Bacterianas , Antibacterianos/farmacología , Fitoquímicos , Extractos Vegetales/farmacología , Pruebas de Sensibilidad Microbiana
9.
Anim Biotechnol ; 34(7): 2441-2448, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-35792780

RESUMEN

A cost effective, simple and rapid method is critical for detection of porcine circovirus type 2 (PCV2) infection in pigs. The present study reports the development and evaluation a loop-mediated isothermal amplification (LAMP) assay for rapid visual detection of PCV2 in pigs. The time and temperature conditions for amplification of PCV2 genes were optimized to be 30 min at 67 °C. The developed assay was 10 fold more sensitive than conventional PCR with analytical sensitivity of 5 pg and 50 pg, respectively. The developed LAMP assay had a sensitivity of 100%, specificity of 85.45% and overall accuracy of 89.70%. This is perhaps the most rapid of all LAMP reports for PCV2 detection available globally. The assay did not cross-react with porcine parvovirus or classical swine fever virus. DNA sequencing was done to ensure accuracy of LAMP assay results. The assay was assembled into a kit of 20 reactions and validated in different laboratories in India. The developed LAMP assay was proved to be a specific, sensitive and rapid method for visual detection of PCV2 which does not require costly equipments.


Asunto(s)
Circovirus , Enfermedades de los Porcinos , Animales , Porcinos , Enfermedades de los Porcinos/diagnóstico , Circovirus/genética , Sensibilidad y Especificidad , Técnicas de Amplificación de Ácido Nucleico/veterinaria , Técnicas de Amplificación de Ácido Nucleico/métodos
10.
Anim Biotechnol ; 34(6): 1883-1890, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-35343866

RESUMEN

A diagnostic method for simultaneously detecting and distinguishing African Swine Fever (ASF), porcine circovirus type 2 (PCV2), and porcine parvovirus (PPV) in clinical specimens is critical for differential diagnosis, monitoring, and control in the field. Three primer pairs were designed and used to create a multiplex PCR assay. In addition, 356 porcine post mortem tissue samples from various parts of India's North Eastern region were tested by the developed multiplex PCR assay to demonstrate its accuracy. Using the designed primers, each of the ASF, PCV2 and PPV target genes was amplified, but no other porcine virus genes were detected. The assay's limit of detection was 102 copies/µl of PCV2, PPV, or ASFV. The detection of PCV2, PPV, and ASF in postmortem tissue samples revealed that they are co-circulating in India's North-Eastern region. The percentage positivity (PP) for PCV2, PPV and ASF single infection were 7.02% (25/356), 3.93% (14/356), and 3.37% (12/356), respectively, while the PP for PCV2& PPV co-infection was 2.80% (10/356), ASF & PCV2 co infection was 1.4% (5/356) and the ASF, PPV& PCV2 co-infection was1.40% (5/356). The results also indicate that the ASF can infect pigs alongside PCV and PPV.


Asunto(s)
Fiebre Porcina Africana , Infecciones por Circoviridae , Coinfección , Infecciones por Parvoviridae , Parvovirus Porcino , Enfermedades de los Porcinos , Virosis , Animales , Porcinos , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , Reacción en Cadena de la Polimerasa Multiplex/métodos , Fiebre Porcina Africana/diagnóstico , Coinfección/diagnóstico , Coinfección/veterinaria , Infecciones por Circoviridae/diagnóstico , Infecciones por Circoviridae/veterinaria , Enfermedades de los Porcinos/diagnóstico , Virosis/diagnóstico , Parvovirus Porcino/genética
11.
Anim Biotechnol ; 34(8): 3946-3961, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37587839

RESUMEN

Muscle development is an important priority of pig breeding programs. There is a considerable variation in muscularity between the breeds, but the regulation mechanisms of genes underlying myogenesis are still unclear. Transcriptome data from two breeds of pigs with divergent muscularity (Mali and Hampshire) were integrated with histology, immunofluorescence and meat yield to identify differences in myogenesis during the early growth phase. The muscle transcriptomics analysis revealed 17,721 common, 1413 and 1115 unique transcripts to Hampshire and Mali, respectively. This study identified 908 differentially expressed genes (p < 0.05; log2FC > ±1) in the muscle samples, of which 550 were upregulated and 358 were downregulated in Hampshire pigs, indicating differences in physiological process related to muscle function and development. Expression of genes related to myoblast fusion (MYMK), skeletal muscle satellite cell proliferation (ANGPT1, CDON) and growth factors (HGF, IGF1, IGF2) were higher in Hampshire than Mali, even though transcript levels of several other myogenesis-related genes (MYF6, MYOG, MSTN) were similar. The number of fibers per fascicle and the expression of myogenic marker proteins (MYOD1, MYOG and PAX7) were more in Hampshire as compared to Mali breed of pig, supporting results of transcriptome studies. The results suggest that differences in muscularity between breeds could be related to the regulation of myoblast fusion and myogenic activities. The present study will help to identify genes that could be explored for their utility in the selection of animals with different muscularities.


Asunto(s)
Sus scrofa , Transcriptoma , Porcinos/genética , Animales , Transcriptoma/genética , Sus scrofa/genética , Músculo Esquelético/metabolismo , Malí , Regulación del Desarrollo de la Expresión Génica , Desarrollo de Músculos/genética
12.
Molecules ; 28(20)2023 Oct 10.
Artículo en Inglés | MEDLINE | ID: mdl-37894491

RESUMEN

Staphylococcus aureus is a common human pathogen. Methicillin-resistant Staphylococcus aureus (MRSA) infections pose significant and challenging therapeutic difficulties. MRSA often acquires the non-native gene PBP2a, which results in reduced susceptibility to ß-lactam antibiotics, thus conferring resistance. PBP2a has a lower affinity for methicillin, allowing bacteria to maintain peptidoglycan biosynthesis, a core component of the bacterial cell wall. Consequently, even in the presence of methicillin or other antibiotics, bacteria can develop resistance. Due to genes responsible for resistance, S. aureus becomes MRSA. The fundamental premise of this resistance mechanism is well-understood. Given the therapeutic concerns posed by resistant microorganisms, there is a legitimate demand for novel antibiotics. This review primarily focuses on PBP2a scaffolds and the various screening approaches used to identify PBP2a inhibitors. The following classes of compounds and their biological activities are discussed: Penicillin, Cephalosporins, Pyrazole-Benzimidazole-based derivatives, Oxadiazole-containing derivatives, non-ß-lactam allosteric inhibitors, 4-(3H)-Quinazolinones, Pyrrolylated chalcone, Bis-2-Oxoazetidinyl macrocycles (ß-lactam antibiotics with 1,3-Bridges), Macrocycle-embedded ß-lactams as novel inhibitors, Pyridine-Coupled Pyrimidinones, novel Naphthalimide corbelled aminothiazoximes, non-covalent inhibitors, Investigational-ß-lactam antibiotics, Carbapenem, novel Benzoxazole derivatives, Pyrazolylpyridine analogues, and other miscellaneous classes of scaffolds for PBP2a. Additionally, we discuss the penicillin-binding protein, a crucial target in the MRSA cell wall. Various aspects of PBP2a, bacterial cell walls, peptidoglycans, different crystal structures of PBP2a, synthetic routes for PBP2a inhibitors, and future perspectives on MRSA inhibitors are also explored.


Asunto(s)
Staphylococcus aureus Resistente a Meticilina , Humanos , Proteínas de Unión a las Penicilinas/química , Staphylococcus aureus Resistente a Meticilina/metabolismo , Meticilina/metabolismo , Meticilina/farmacología , Staphylococcus aureus/metabolismo , Antibacterianos/farmacología , Antibacterianos/metabolismo , Monobactamas/metabolismo , Proteínas Bacterianas/química , Pruebas de Sensibilidad Microbiana
13.
Indian J Clin Biochem ; 38(2): 251-261, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-37025430

RESUMEN

Reserpine, a bioactive compound isolated from the roots of Rauwolfia serpentine, is known to deplete dopamine, a neurotransmitter. The clinical application of reserpine has been associated to manage hypertension, insanity, insomnia and schizophrenia. However, the usage of reserpine as a drug is restricted because of its ability of inducing excess free radicals production and oxidative stress resulting into damage to liver and other organs. Here, we have explored the antioxidative potential of extract of garlic prepared using ethanol (EEG) against reserpine-induced hepatic damage in the albino Wister rats.The animals were divided into four different groups containing 6 animals in each: (1) control + placebo, (2) control + EEG, (3) reserpine and (4) reserpine with EEG. The reserpine treatment resulted into sharp increase in the level of MDA and significant reduction in the activitiesof key antioxidative enzymes (SOD, GST, and CAT) in the rat liver. It also caused sharp perturbations in the levels of certain hepatic transaminases (ALT, AST) and glycolytic LDH. The histopathological results revealed hepatic necrosis, which could have occurred due to reserpine induced lipid peroxidation as well as reduction in the levels of antioxidant species.The administration of EEG, however, significantly ameliorated reserpine induced hepatotoxicity. These results reflected the ameliorative property of EEG, which was probably mediated via its antioxidant function as it contains several bioactive molecules with free radical quenching potential.This study suggestedthe prospective application of EEG as a supplement to combat the side effects of reserpine.

14.
J Food Sci Technol ; 60(4): 1274-1283, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36936116

RESUMEN

Foodborne illnesses are leading source of morbidity and mortality in both developed and developing nations. Escherichia coli O157 is one of the most reported foodborne pathogen that emerged in the past few decades. South East Asia region suffers the highest average burden of diarrhoeal mortality, especially when it comes to child mortality.Query Many studies were undertaken in the developed nations to evaluate the role of E. coli O157 as one of the etiological agent in foodborne outbreaks. In this article, we discuss the distribution of E. coli O157 serotype in the food chains of South East Asian countries, with a special focus on India where more than half a million child diarrhoeal deaths occurs every year and the reasons for which is often not ascertained to the fullest extent. The article also describes in detail about the various detection methods and control measures with respect to E. coli O157. The aim of this study is to document and highlight the extent of Foodborne infections of E. coli O157 origin and thereby taking effective and proactive preventive measures.

15.
J Food Sci Technol ; 60(1): 132-146, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-36618039

RESUMEN

In order to ensure food safety, screening food samples for the presence of pathogens has been categorised as a legal testing item throughout the globe. One of the most prevalent zoonotic bacteria transmitted through dairy milk is Staphylococcus aureus. Given the limitations of the conventional detection methods, in the current study we desigined a competitive lateral flow immune assay (LFIA) using colloidal silver nanoparticles derived from mango leaves for the detection of Staphylococcus aureus in cow milk. SpA, a recombinant protein of Staphylococcus aureus, was used to raised hyperimmune sera used for developing the assay followed by conjugation with the synthesized nanoparticles. To increase the specificity of the assay, the milk samples were prenriched with selective agar exclusively require for Staphyloccocus aureus. The assay was found to be completed within 7-8 h by observing test and control lines in LFIA strips. The developed assay was found to specifically detect the bacteria as low as 1000 cfu/ml of milk samples. With a total 230 number of raw and clinical mastitis milk samples, the assay was validated and achieved relative accuracy, specificity, and sensitivity values of 97.39, 98.03, and 96.1%, respectively. The developed LFIA, which uses economically feasible and stable silver nanoparticles derived from mango leaves, has the potential for routine screening of milk samples for the presence of Staphylococcus aureus, especially in low-resource settings, allowing for early diagnosis, which facilitates effective treatment for the dairy animals and prevents the transmission of the disease in consumers.

16.
Infect Immun ; 90(5): e0006022, 2022 05 19.
Artículo en Inglés | MEDLINE | ID: mdl-35420446

RESUMEN

Brucella species are intracellular bacterial pathogens, causing the worldwide zoonotic disease brucellosis. Brucella invades professional and nonprofessional phagocytic cells, followed by resisting intracellular killing and establishing a replication permissive niche. Brucella also modulates the innate and adaptive immune responses of the host for its chronic persistence. The complex intracellular cycle of Brucella depends in a major way on multiple host factors, but limited information is available on host and bacterial proteins that play an essential role in the invasion, intracellular replication, and modulation of host immune responses. By employing a small interfering RNA (siRNA) screening, we identified a role for the host protein FBXO22 in the Brucella-macrophage interaction. FBXO22 is the key element in the SCF E3 ubiquitination complex, where it determines the substrate specificity for ubiquitination and degradation of various host proteins. Downregulation of FBXO22 by siRNA or the CRISPR-Cas9 system resulted in diminished uptake of Brucella into macrophages, which was dependent on NF-κB-mediated regulation of phagocytic receptors. FBXO22 expression was upregulated in Brucella-infected macrophages, which resulted in induction of phagocytic receptors and enhanced production of proinflammatory cytokines through NF-κB. Furthermore, we found that FBXO22 recruits the effector proteins of Brucella, including the anti-inflammatory proteins TcpB and OMP25, for degradation through the SCF complex. We did not observe any role for another F-box-containing protein of the SCF complex, ß-TrCP, in the Brucella-macrophage interaction. Our findings unravel novel functions of FBXO22 in host-pathogen interaction and its contribution to pathogenesis of infectious diseases.


Asunto(s)
Brucella , Brucelosis , Proteínas F-Box , Antiinflamatorios/metabolismo , Brucella/metabolismo , Brucelosis/microbiología , Citocinas/metabolismo , Preparaciones de Acción Retardada/metabolismo , Proteínas F-Box/genética , Proteínas F-Box/metabolismo , Humanos , Macrófagos , FN-kappa B/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Receptores Citoplasmáticos y Nucleares/metabolismo
17.
Adv Funct Mater ; 32(47)2022 Nov 17.
Artículo en Inglés | MEDLINE | ID: mdl-36816792

RESUMEN

Vascular endothelial cell (EC) plasticity plays a critical role in the progression of atherosclerosis by giving rise to mesenchymal phenotypes in the plaque lesion. Despite the evidence for arterial stiffening as a major contributor to atherosclerosis, the complex interplay among atherogenic stimuli in vivo has hindered attempts to determine the effects of extracellular matrix (ECM) stiffness on endothelial-mesenchymal transition (EndMT). To study the regulatory effects of ECM stiffness on EndMT, an in vitro model is developed in which human coronary artery ECs are cultured on physiological or pathological stiffness substrates. Leveraging single-cell RNA sequencing, cell clusters with mesenchymal transcriptional features are identified to be more prevalent on pathological substrates than physiological substrates. Trajectory inference analyses reveal a novel mesenchymal-to-endothelial reverse transition, which is blocked by pathological stiffness substrates, in addition to the expected EndMT trajectory. ECs pushed to a mesenchymal character by pathological stiffness substrates are enriched in transcriptional signatures of atherosclerotic ECs from human and murine plaques. This study characterizes at single-cell resolution the transcriptional programs that underpin EC plasticity in both physiological or pathological milieus, and thus serves as a valuable resource for more precisely defining EndMT and the transcriptional programs contributing to atherosclerosis.

18.
Soft Matter ; 18(37): 7201-7216, 2022 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-36098333

RESUMEN

Nanoscale self-assembly of peptide constructs represents a promising means to present bioactive motifs to develop new functional materials. Here, we present a series of peptide amphiphiles which form hydrogels based on ß-sheet nanofibril networks, several of which have very promising anti-microbial and anti-parasitic activities, in particular against multiple strains of Leishmania including drug-resistant ones. Aromatic amino acid based amphiphilic supramolecular gelators C14-Phe-CONH-(CH2)n-NH2 (n = 6 for P1 and n = 2 for P3) and C14-Trp-CONH-(CH2)n-NH2 (n = 6 for P2 and n = 2 for P4) have been synthesized and characterized, and their self-assembly and gelation behaviour have been investigated in the presence of ultrapure water (P1, P2, and P4) or 2% DMSO(v/v) in ultrapure water (P3). The rheological, morphological and structural properties of the gels have been comprehensively examined. The amphiphilic gelators (P1 and P3) were found to be active against both Gram-positive bacteria B. subtilis and Gram-negative bacteria E. coli and P. aeruginosa. Interestingly, amphiphiles P1 and P3 containing an L-phenylalanine residue show both antibacterial and antiparasitic activities. Herein, we report that synthetic amphiphiles with an amino acid residue exhibit a potent anti-protozoan activity and are cytotoxic towards a wide array of protozoal parasites, which includes Indian varieties of Leishmania donovani and also kill resistant parasitic strains including BHU-575, MILR and CPTR cells. These gelators are highly cytotoxic to promastigotes of Leishmania and trigger apoptotic-like events inside the parasite. The mechanism of killing the parasite is shown and these gelators are non-cytotoxic to host macrophage cells indicating the potential use of these gels as therapeutic agents against multiple forms of leishmaniasis in the near future.


Asunto(s)
Aminoácidos , Antiinfecciosos , Antibacterianos/química , Antibacterianos/farmacología , Antiparasitarios/farmacología , Dimetilsulfóxido , Escherichia coli , Hidrogeles/química , Hidrogeles/farmacología , Péptidos/química , Fenilalanina , Pseudomonas aeruginosa , Agua
19.
Appl Opt ; 61(27): 8057-8063, 2022 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-36255927

RESUMEN

The structure of silica single-mode fiber (SMF) must be modified in order to develop optical fiber-based biosensors. To reduce the diameter of the optical fiber, a low-cost chemical etching method is very popular. The proposed chemical etching method is a simple, rapid, and cost-effective technique for removing the silica cladding up to a desired diameter. In the laboratory, hydrofluoric acid (HF acid, 40% concentration) is used for etching. A variation on etching is also proposed and tested with 40% HF as well as with magnetic stirring at the different speeds. The etching experiments are also carried out at different temperatures. The etching results of silica fiber are presented through a step-by-step procedure using a rapid and resource-efficient method for the fabrication of optical fiber-based biosensors. The etched diameter characterization is done using a calibrated compound microscope. The sensing experiment with unetched and etched optical fiber is also performed for the detection of different concentrations of glucose biomolecules.


Asunto(s)
Técnicas Biosensibles , Fibras Ópticas , Ácido Fluorhídrico/química , Dióxido de Silicio , Glucosa
20.
Eur J Wildl Res ; 68(5): 59, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35992994

RESUMEN

We report an incidence of natural infection of SARS-CoV-2 in free-ranging Indian leopard (Panthera pardus fusca). The case was detected during routine screening. Post-mortem and laboratory examination suggested virus-induced interstitial pneumonia. Viral genome could be detected in various organs including brain, lung, spleen, and lymph nodes by real-time PCR. Whole-genome sequence analysis confirmed infection of Pango lineage B.1.617.2 of SARS-CoV-2. Till now, only Asiatic lions have been reported to be infected by SARS-CoV-2 in India. Infections in animals were detected during peak phase of pandemic and all the cases were captive with close contacts with humans, whereas the present case was observed when human cases were significantly low. No tangible evidence linked to widespread infection in the wild population and the incidence seems to be isolated case. High nucleotide sequence homology with prevailing viruses in humans suggested spillover infection to the animal. This report underlines the need for intensive screening of wild animals for keeping track of the virus evolution and development of carrier status of SARS-CoV-2 among wildlife species. Supplementary Information: The online version contains supplementary material available at 10.1007/s10344-022-01608-4.

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