An insight into whole genome sequencing data of methicillin-resistant Staphylococcus aureus circulating in a teaching hospital in North India.

PURPOSE: To provide an insight into the Whole-genome sequencing (WGS) data of MRSA strains circulating in a teaching hospital in north India. METHODS: An exploratory study was conducted in which fifty non-repetitive MRSA isolates obtained from pus samples of inpatients from July 2018 to February 2019 were subjected to preliminary identification (ID) and antibiotic susceptibility testing (AST) at our centre. These isolates were later sent to Central Research Laboratory, India for further testing using the VITEK-2 compact system followed by WGS. Only eighteen isolates were eventually considered for final analysis and the rest (n â€‹= â€‹32) were excluded due to various technical reasons. RESULTS: The WGS confirmed MRSA isolates predominantly belonged to CC22 (56.25%) and CC30 (31.25%). The CC22 MRSA strains carried SCCmec types IVa (77.8%) & IVc (22.2%) and belonged to spa types t005 (44.4%), t4584 (22.2%), t11808 (11.1%), t1328 (11.1%) and t309 (11.1%), respectively. The MRSA isolates of CC30 carried SCCmec types IVa (60%), IVg (20%) & V (20%) and belonged to spa types t021 (80%) & t2575 (20%), respectively. One MRSA isolate carried a novel SCCmec type V. The luk-PV and tsst-1 genes were present in 93.75% and 33.33% of MRSA isolates, respectively. The concordance between the phenotypic and genotypic AST results was 100% for Beta-lactams, Fluoroquinolones, Tetracyclines & Lipoglycopeptides, respectively. CONCLUSIONS: Through this study, we intend to embark upon a relatively newer avenue of clinical-genomic surveillance of nosocomial bacterial isolates like MRSA, which would help us improve the existing infection control and antibiotic stewardship practices in our hospital.

Incidence of SARS-CoV-2 Co-infections During the Second Wave in Sub-Himalayan Region, India.

Introduction The second wave of the coronavirus disease 2019 (COVID-19) pandemic in India, which started from April 2021, has been more severe and deadly than the first wave. The aim of this prospective study was to determine the possibility of other respiratory pathogens contributing towards the severity and hospitalization in the current second wave. Materials and methods Nasopharyngeal and oropharyngeal swab samples were collected and processed for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by reverse transcription polymerase chain reaction (RT-PCR). These samples were further processed for detection of co-infection in SARS CoV-2 patients by BioFire® Filmarray® 2.0 (bioMérieux, USA). Results We screened 77 COVID-19-positive patients admitted to All India Institute of Medical Sciences (AIIMS), Rishikesh and found cases of co-infections in five (6.49 %) patients. Conclusion Our finding suggests that co-infections had no or minimal role in augmenting the second wave of the COVID-19 pandemic in India, and the emergence of new variants may be the probable cause.

Elucidation of the signalling pathways for enhanced exosome release from Mycobacterium-infected macrophages and subsequent induction of differentiation.

Exosomes are extracellular vesicles released by all cell types; perform several important functions such as cell-to-cell communication, growth, differentiation and so on. Exosomes elicit several signalling mechanisms as they carry information in the form of DNA, RNA or protein docked on them. We show that exosomes released from Mycobacterium tuberculosis (Mtb)-infected macrophages not only induce differentiation of naïve monocytes but also generate functionally active macrophages via MAPK-dependent signalling mechanism through MK-2 and NF-κß activation which is completely different from the differentiation induced by exosomes from uninfected macrophages. Further, we elucidate unequivocally the signalling mechanism behind the enhanced release of exosome generation from infected macrophages driven by AKT phosphorylation involving Rab7a and Rab11a. Genes of both ESCRT-dependent and -independent pathways are found to be involved in enhanced exosomes release and are modulated by AKT. However, interestingly, the genes of the ESCRT-independent pathway are dependent on NF-κß activation while the genes of the dependent pathway are not, suggesting two parallel signalling cascades operating in tandem.

Microscopic Parotidectomy: A Prospective Study.

To assess the parameters' setting of the microscope during parotidectomy and the impact of microscopic parotidectomy on facial nerve functional status. A prospective study was conducted on 28 patients in a tertiary care center, who underwent microscopic parotidectomy. Microscope's settings' like magnification, focal length, diameter of the visualized field, and clock position were recorded. Facial nerve functional status was also recorded. All surgeries were performed by right-handed surgeons using Leica F 20 M525 microscope. Clock position of microscope for right parotidectomy ranged between 7 and 10 o clock and for left, it ranged between 7 and 12 o clock. Magnification ranged between 1.3 × and 3.2 ×; magnifications of 1.3 × and 1.8 × were preferred from incision to separation of parotid from sternocleidomastoid muscle, 1.8 × and 2 × for dissection of the facial nerve trunk, and 2 × and 3.2 × for individual branches of the facial nerve. Focal length ranged between 251 and 410 mm and the diameter of the visualized field ranged between 7 and 14.7 cm. Out of 24 cases of benign lesions, 2 (8.3%) developed facial paresis which resolved in 3 months. Two out of four cases of malignancy developed permanent palsy as nerve branches were sacrificed to achieve tumor clearance. Using a microscope for parotidectomy is advantageous for facial nerve dissection, reducing chances of facial paralysis. The microscope was also found to be useful for teaching. The use of appropriate microscopic parameters avoids the glitch of transition from open to microscopic approach.

Novel Modifications to Total Maxillary Swing Approach in Extensive Nasopharyngeal Angiofibroma to Minimize Complications.

Maxillary swing approaches provide excellent exposure of the anterior, mid, and anterolateral skull base offering a wide window to approach nasopharyngeal neoplasms; however, they are also associated with complications. The present study aimed to evaluate the results of a modified total maxillary swing (TMS) approach developed to minimize postoperative complications. The modified TMS approach was used to treat five patients who had extensive juvenile nasopharyngeal angiofibromas between March and October 2019 at our tertiary care center. Surgical technique, preoperative image findings, and intra-operative findings were recorded. In the postoperative follow-up, patients were examined to rule out all possible complications associated with the procedure according to the literature. A retrospective analysis was performed to assess tumor extensions, surgical modifications, and postoperative complications. All tumors had orbital and infratemporal (lateral limit) involvement while four had intracranial involvement. No per-operative complications were reported, and postoperative clinical and endoscopic evaluation was performed at 1, 2, and 3 months. There was no evidence of complications including residue, recurrence, maxillary necrosis, ophthalmoplegia, epiphora, palatal fistula, or jaw malocclusion. Besides, minor complications such as infraorbital margin skin retraction, infraorbital serous collection, maxillo-zygomatic abscess, and unsightly scar were also not seen. Only one case presented with maxillary osteomyelitis which was resolved with mini-plate removal and antibiotics. A modified TMS approach is a prudent option to ensure complete removal of juvenile nasopharyngeal angiofibromas with negligible complications.

Amino Acid-triggered Water-soluble NBD Derivatives for Differential Organelle Staining and the Role of the Chemical Moiety for their Specific Localization.

Apart from being the unit of proteins, amino acids have diverse roles. Here we have shown that amino acids guide the differential transportation of a dye molecule to the cellular organelles depending upon the property of their intrinsic functionality. We have conjugated the nitrobenzofurazan (NBD) moiety with two amino acids (lysine and histidine derivatives) with a linker. Both derivates are water-soluble and biocompatible in nature. Surprisingly, we found that lysine-conjugated NBD (NBD-Lys) stains lipid droplets whereas histidine-conjugated NBD (NBD-His) stains lysosomes. We also measured the spectral properties of these two NBD conjugates. The results depict that both conjugates are extremely stable both in air and under inert atmosphere, and the fluorescence of the derivatives remains almost unaltered at different pH. Further by altering the functionality of the side chain, we established the contribution of each functional group towards this differential organelle targeting.

Comparative Evaluation of Colistin Broth Disk Elution Method With Two Commercially Available Systems for Colistin Susceptibility Testing Against Carbapenem-Resistant Klebsiella pneumoniae: A Single-Center Exploratory Study.

PURPOSE: The purpose is to explore the diagnostic utility of colistin broth disk elution (CBDE) as a simple and reliable method of colistin susceptibility testing. MATERIALS AND METHODS: An exploratory study was undertaken in a tertiary care teaching hospital in Uttarakhand, from September 2021 to March 2022, after obtaining approval from the Institute Ethics Committee. Twenty-five non-repetitive carbapenem-resistant Klebsiella pneumoniae clinical isolates were included in the study. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and BD Phoenix M50 system were used to perform species-level identification and antibiotic susceptibility testing (AST), respectively, as per the manufacturer's instructions. AST results (including those of colistin) were interpreted as per the CLSI guidelines 2022. The test isolates were further subjected to additional in vitro colistin susceptibility testing using a commercially available Mikrolatest colistin susceptibility testing kit and CBDE, respectively. RESULTS: The in vitro colistin resistance rates varied from 8% by BD Phoenix system to 20% by Mikrolatest kit and 32% by CBDE, respectively. For colistin susceptibility, a higher CA was observed between the BD Phoenix system and CBDE (64.71%) than between the Mikrolatest kit and CBDE (31.60%). Overall, a statistically significant fair agreement was observed between the BD Phoenix system and CBDE (Kappa: 0.312; 95% CI: 0.036 to 0.660) and Mikrolatest MIC colistin kit and CBDE (Kappa: 0.286; 95% CI: 0.111 to 0.683), respectively. CONCLUSIONS: In vitro colistin testing remains a significant challenge globally. Although the present study results are inconclusive due to the small sample size, we should conduct multi-centric studies globally, taking a considerable sample size representing different Gram-negative bacilli to generate conclusive evidence on the utility of CBDE as a reliable method of colistin susceptibility testing.

The Mystery of Mucormycosis in COVID-19: A Multifactorial Menace or an Enigmatic Delta Variant Associated Phenomenon? An Exploratory Study from a Tertiary Care Centre in North India with a Brief Literature Review.

Background: There was a global surge in cases of mucormycosis in COVID-19 patients during the second wave of the pandemic in 2021, reported especially from India. Various predisposing factors such as diabetes mellitus, rampant use of corticosteroids, and COVID-19 per se may be responsible for this spike. Some public health experts have postulated that the epidemiological link between the Delta variant of SARS-CoV-2 and mucormycosis should be explored. Material and Methods: A retrospective exploratory study was conducted, in which data of 15 laboratory-confirmed cases of COVID-19 with mucormycosis and/or aspergillosis co-infections were collected after obtaining approval from the institute's ethics committee. These patients were admitted to the Mucor wards of our hospital. The positive COVID-19 status of these patients was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR). The residual SARS-CoV-2 RNA containing elutes of these patients were stored at -80°C in deep freezers and subjected to whole-genome sequencing in June 2021 at the National Centre for Disease Control (NCDC), New Delhi, India as part of the Indian SARS-CoV-2 Genomic Consortia (INSACOG) program. Concomitant fungal infections in these patients were diagnosed by KOH wet mount and fungal culture as per standard guidelines. Descriptive statistics in the form of percentages and median were used to report the findings. Results: Periorbital swelling and ocular pain (14/15; 93.33%), followed by facial swelling (11/15; 73.33%) and nasal obstruction (9/15; 60%), were the most common clinical features observed in these patients. Rhizopus arrhizus was the most common causative fungal agent (12/15; 80%). The majority of the patients (9/13; 69.23%) were infected with the Delta variant of SARS-CoV-2. Conclusion: COVID-associated mucormycosis seems to be multifactorial in origin. Although there may be a possible association between mucormycosis and the Delta variant, more studies should be conducted to explore this seemingly reasonable proposition.

Renaissance of Hansen's Disease in Post-Elimination Era in North India: A Retrospective Clinico-Bacteriological Study.

Introduction Hansen's disease is a chronic infectious disease caused by Mycobacterium leprae. India declared the elimination of leprosy in December 2005, but a slow resurgence of the disease still continues in several parts of India. The diagnosis of leprosy is primarily clinical but slit-skin smear microscopy aids in an accurate diagnosis. There are very few studies on clinico-bacteriological patterns of leprosy at this post-elimination phase. Aim This study aimed to analyze the clinical and bacteriological findings of newly diagnosed cases of Hansen's disease in the post-elimination era. Materials and methods This is a descriptive, hospital-based, retrospective study of newly diagnosed cases of Hansen's disease, enrolled in the Hansen's disease clinic attached to the dermatology outpatient department (OPD) of a tertiary care hospital in North India. A retrospective chart review of newly diagnosed cases of leprosy for a period of one year was done. Information about demographics, clinical characteristics, spectrum of disease, and slit skin smear data of patients were collected. Statistical analysis was performed using SPSS Version 16.0 (Chicago, IL, SPSS Inc.). Result A total of 116 patients were included of which 68.1% (79) were males. The age of patients ranged from 7 to 72 years and children (<15 years) constituted 6% (7/116) of all cases. The most common clinical spectrum was borderline lepromatous leprosy 37.9% (44/116) followed by lepromatous leprosy 32.8% (38/116). Out of 116 cases, 39.6% of cases showed slit-skin smear positivity. Conclusion The study brings forth evidence on the slow re-emergence of leprosy in India. In this study, multibacillary cases outnumber the paucibacillary cases; also, childhood cases were encountered indicating active community spread of the disease in the "post-elimination era." There is an urgent need to step up the surveillance for Hansen's disease to curb the further spread of the bacilli in the community.

Delineating the complex mechanistic interplay between NF-κß driven mTOR depedent autophagy and monocyte to macrophage differentiation: A functional perspective.

Autophagy is an extremely essential cellular process aimed to clear redundant and damaged materials, namely organelles, protein aggregates, invading pathogens, etc. through the formation of autophagosomes which are ultimately targeted to lysosomal degradation. In this study, we demonstrated that mTOR dependent classical autophagy is ubiquitously triggered in differentiating monocytes. Moreover, autophagy plays a decisive role in sustaining the process of monocyte to macrophage differentiation. We have delved deeper into understanding the underlying mechanistic complexities that trigger autophagy during differentiation. Intrigued by the significant difference between the protein profiles of monocytes and macrophages, we investigated to learn that autophagy directs monocyte differentiation via protein degradation. Further, we delineated the complex cross-talk between autophagy and cell-cycle arrest in differentiating monocytes. This study also inspects the contribution of adhesion on various steps of autophagy and its ultimate impact on monocyte differentiation. Our study reveals new mechanistic insights into the process of autophagy associated with monocyte differentiation and would undoubtedly help to understand the intricacies of the process better for the effective design of therapeutics as autophagy and autophagy-related processes have enormous importance in human patho-physiology.

Comparative Evaluation of Nasopharyngeal and Oropharyngeal Swab Based Rapid SARS-CoV-2 Antigen Detection and Real-Time RT-PCR for Diagnosis of COVID-19 in Tertiary Care Hospital.

PURPOSE: To assess and compare the diagnostic accuracy of the GenBody COVID-19 Antigen kit (GenBody Inc., Cheonan, South Korea) available in the market with the gold standard reverse transcription-polymerase chain reaction (RT-PCR) assay to detect severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). METHODS: Nasopharyngeal and oropharyngeal swabs were collected from suspected coronavirus disease 2019 (COVID-19) patients and tested by RT-PCR and GenBody Rapid antigen kit. Performance characteristic of the antigen kit was calculated. RESULTS: We tested nasopharyngeal swabs and oropharyngeal swabs (n=240). Amongst the 102 positive RT-PCR samples, the rapid antigen test detected 36 as positive, showing an overall sensitivity of 35.3%. All the samples detected positive with the antigen rapid test were also detected positive by RT-PCR. CONCLUSION: The performance of the rapid antigen kit was good with respect to high viral load samples, whereas those with lower levels were missed. Unfortunately, the overall low sensitivity of the antigen kit does not allow using it alone as the frontline testing kit for COVID-19 diagnosis.

Surviving COVID-19 and multiple complications post total laryngectomy.

The clinical manifestation of novel COVID-19 is variable. Pre-existing carcinoma and other comorbidities have been associated with increased COVID-19-related morbidity and mortality. Surgical intervention for advanced laryngeal carcinoma in old age during the COVID-19 pandemic may pose multiple challenges to the patient and the treatment team. We report a case of a 67-year-old elderly man who developed SARS-CoV-2 infection on the 21st day following total laryngectomy and neck dissection. The postoperative period was complicated by sequential development of pulmonary embolism, neck infection, pharyngeal leak and COVID-19 which were managed successfully. No close contacts were positive on the reverse transcription-PCR test for SARS-CoV-2. The patient is in follow-up for the past 7 months without any recurrence or COVID-19-related morbidity. The successful recovery and no cross-infection may be attributed to early diagnosis, immediate intervention and properly implemented institutional infection control policy.

Lipid droplet: A functionally active organelle in monocyte to macrophage differentiation and its inflammatory properties.

Lipid droplets (LDs) perform several important functions like inflammatory responses, membrane trafficking, acts as secondary messengers, etc. rather than simply working as an energy reservoir. LDs have been implicated as a controlling factor in the progression of atherosclerosis followed by foam cell formation that derives from macrophages during the differentiation process. However, the role of LDs in monocyte differentiation or its further immunological function is still an area that mandates in-depth investigation. We report that LD dynamics is important for differentiation of monocytes and is absolutely required for sustained and prolonged functional activity of differentiated macrophages. In THP-1 cell line model system, we elucidated that increase in total LD content in monocyte by external lipid supplements, can induce monocyte differentiation independent of classical stimuli, PMA. Differential expression of PLIN2 and ATGL during the event, together with abrogation of de novo lipogenesis further confirmed the fact. Besides, an increase in LD content by free fatty acid supplement was able to exert a synergistic effect with PMA on differentiation and phagocytic activity compared to when they are used alone. Additionally, we have shown Rab5a to play a vital role in LDs biosynthesis/maturation in monocytes and thereby directly affecting differentiation of monocytes into macrophages via AKT pathway. Thus our study reveals the multi-faceted function of LDs during the process of monocyte to macrophage differentiation and thereby helping to maintain the functional activity.

Candida auris emergence in the Himalayan foothills: First case report from Uttarakhand, India.

BACKGROUND AND PURPOSE: Candida auris is a rapidly emerging fungus, which is considered globally a cause of concern for public health. This report describes the first case of C. auris fungemia from a tertiary care hospital in the hilly state of Uttarakhand in India. CASE REPORT: The patient was a 37-year-old female who underwent a Whipple procedure for the carcinoma of the head of the pancreas. She developed fever 12 days after the operation while recovering from surgery in the hospital. Blood culture yielded C. auris which was identified by the matrix-assisted laser desorption/ionization-time of flight mass spectrometry (Bruker Daltonics, Germany). The patient was successfully treated with caspofungin. CONCLUSION: In conclusion, C. auris is potentially multidrug resistant, resulting in nosocomial outbreaks and sporadic infections which can be potentially prevented when identified early by implementing contact precautions.

Bacteriological analysis of street-vended fruit juices available in Rishikesh, Uttarakhand.

BACKGROUND: A descriptive study was carried out due to absence of knowledge of relation between hygiene status of vendor and extent of contamination while simultaneously adding to the pool of similar studies, which would in turn help government formulate an appropriate policy. METHODOLOGY: A sample size of 80 fruit juices were collected from 55 vendors and cultured using CLED agar; relevant biochemical tests were run and isolates confirmed using Chromogenic Agar CPS3Id. The CFU of isolates obtained from one vendor were averaged to obtain average CFU for the vendor. The vendors were interviewed and investigator's observations were also recorded. Both were used to formulate an ingenious hygiene score scale, using which vendor hygiene score (VHS) was calculated. VHS and average CFU for the vendor were statistically analyzed and Spearman's rho was calculated along with the scatter plot. RESULTS: Klebsiella sp. (59.05%) and non-Candida albicans (14.28%) are the major bacterial and fungal contaminant respectively. The median VHS is -9, which is unsuitable for consumption according to HSS. Spearman's rho was -0.736, suggestive of a statistically significant negative nonlinear correlation between VHS and extent of microbial contamination in street-vended fruit juices. CONCLUSION: The plethora of microbial contamination of street-vended fruit juices is a lucrative dimension for research in fruit juice microbiology to improve general public health, reduce disease mortality and morbidity, reduce economic losses, and to protect the general public from the risk of bioterrorism by stringent surveillance in background of limited employment opportunities and constrained livelihoods of the vendors.

Dipylidium caninum: First case in an adult female from uttarakhand and review of literature.

Dipylidium caninum is a rare cestode infection in India, only previously reported in children. We herein report the case of a 50-year-old female from India who presented with a 1-year history of abdominal pain associated with vomiting, cough, and passing worm segments in stool. She had been treated with albendazole without benefit. A stool examination revealed proglottids and egg clusters of D. caninum. She was treated with praziquental 600 mg once daily for 5 days. At follow-up a month later, she had recovered completely, and repeat stool examination was negative for proglottids and eggs. Dipylidiasis can rarely occur among adults in India and present with abdominal pain and cough.

Triple-negative breast cancer-derived microvesicles transfer microRNA221 to the recipient cells and thereby promote epithelial-to-mesenchymal transition.

The triple-negative phenotype is the most prevalent form of human breast cancer worldwide and is characterized by poor survival, high aggressiveness, and recurrence. Microvesicles (MV) are shredded plasma membrane components and critically mediate cell-cell communication, but can also induce cancer proliferation and metastasis. Previous studies have revealed that protease-activated receptor 2 (PAR2) contributes significantly to human triple-negative breast cancer (TNBC) progression by releasing nano-size MV and promoting cell proliferation, migration, and invasion. MV isolated from highly aggressive human TNBC cells impart metastatic potential to nonmetastatic cells. Over-expression of microRNA221 (miR221) has also been reported to enhance the metastatic potential of human TNBC, but miR221's relationship to PAR2-induced MV is unclear. Here, using isolated MV, immunoblotting, quantitative RT-PCR, FACS analysis, and enzymatic assays, we show that miR221 is translocated via human TNBC-derived MV, which upon fusion with recipient cells, enhance their proliferation, survival, and metastasis both in vitro and in vivo by inducing the epithelial-to-mesenchymal transition (EMT). Administration of anti-miR221 significantly impaired MV-induced expression of the mesenchymal markers Snail, Slug, N-cadherin, and vimentin in the recipient cells, whereas restoring expression of the epithelial marker E-cadherin. We also demonstrate that MV-associated miR221 targets phosphatase and tensin homolog (PTEN) in the recipient cells, followed by AKT Ser/Thr kinase (AKT)/NF-κB activation, which promotes EMT. Moreover, elevated miR221 levels in MV derived from human TNBC patients' blood could induce cell proliferation and metastasis in recipient cells. In summary, miR221 transfer from TNBC cells via PAR2-derived MV induces EMT and enhances the malignant potential of recipient cells.

Salmonella osteomyelitis: A rare extraintestinal manifestation of an endemic pathogen.

Salmonella enterica serovar Typhi and Paratyphi usually cause enteric fever in humans characterized by fever and gastrointestinal symptoms such as diarrhea. Bacteremia is a constant feature of enteric fever, and occasionally, dissemination of bacilli throughout the body results in the establishment of one or more localized foci of persisting infection. This happens especially in patients with preexisting conditions such as hemoglobinopathies, previous joint trauma, surgery, connective tissue diseases, lymphoma, diabetes, alcoholism, immunosuppressive state, and extremes of ages. Rarely, these extraintestinal Salmonella infection cases have also been reported in immunocompetent individuals without any obvious predisposing factors. Salmonella osteomyelitis, a rare sequelae of typhoid, may occur as a result of hematogenous seeding of the bone or joints due to a bacteremic episode. It may also spread from nearby areas of infection or penetrating trauma. Timely diagnosis and treatment of this condition are necessary because if left untreated, it can lead to permanent functional deficits later on. We hereby present a case series with brief review of literature on Salmonella osteomyelitis in immunocompetent patients without any known predisposing conditions over a period of 1 year (2017-2018) from a Tertiary Care Teaching Hospital in Uttarakhand, India.

Protease-activated receptor 2 promotes actomyosin dependent transforming microvesicles generation from human breast cancer.

Apart from blood coagulation, coagulation proteases are involved inextricably in cancer progression/propagation via intra/inter-cellular signaling, mediated predominantly by protease-activated receptors (PARs). Microvesicles (MVs), a plasma membrane shredded component, has recently been identified as an important contributor to human breast cancer metastasis. However, the role of PAR2 in promoting MVs generation from breast cancer cells remains largely unexplored. The objective of this study is to investigate whether coagulation protease-mediated human breast cancer propagation commences via MVs and also to decipher the underlying signaling mechanism. Here, we elicited that coagulation factor-FVIIa and Trypsin activates PAR2, which governs MVs shedding from MDAMB231 cells by altering actomyosin dynamics. Treatment of cells with PAR2 activators facilitate MVs generation by activating three independent (MAPK, P38, and Rho) signaling cascades. MAPK, signals through activating MLCK followed by MLC phosphorylation to alter myosin organization whereas, P38 reorganizes actin dynamics by the sequential activation of MK2 and HSP27. RhoA-dependent ROCK-II activation again contributes to remodeling myosin II activity. Further, both our in vitro and in vivo analyses showed that these MVs potentiate invasive and migratory property to the recipient cells. Breast cancer patients blood show an elevation of TF-bearing, pro-metastatic MVs than normal. These findings give an insight into the detailed signaling mechanism involved in the production of MVs with transforming ability from PAR2-activated human breast cancer cells. Understanding these mechanistic details will certainly help to identify crucial targets for therapeutic interventions in MVs-associated human breast cancer metastasis.