Live-attenuated auxotrophic mutant of <i>Salmonella</i> Typhimurium expressing immunogenic HA1 protein enhances immunity and protective efficacy against H1N1 influenza virus infection

Kamble, Nitin Machindra,Hyoung, Kim Je,Lee, John Hwa Future Medicine 2017 Future microbiology Vol.12 No.9

Intracellular delivery of HA1 subunit antigen through attenuated <i>Salmonella</i> Gallinarum act as a bivalent vaccine against fowl typhoid and low pathogenic H5N3 virus

Kamble, Nitin Machindra,Hyoung, Kim Je,Lee, John Hwa BioMed Central 2017 VETERINARY RESEARCH Vol.48 No.-

<P>Introduction of novel inactivated oil-emulsion vaccines against different strains of prevailing and emerging low pathogenic avian influenza (LPAI) viruses is not an economically viable option for poultry. Engineering attenuated <I>Salmonella</I> Gallinarum (<I>S.</I> Gallinarum) vaccine delivering H5 LPAI antigens can be employed as a bivalent vaccine against fowl typhoid and LPAI viruses, while still offering economic viability and sero-surveillance capacity. In this study, we developed a JOL1814 bivalent vaccine candidate against LPAI virus infection and fowl typhoid by engineering the attenuated <I>S.</I> Gallinarum to deliver the globular head (HA1) domain of hemagglutinin protein from H5 LPAI virus through pMMP65 constitutive expression plasmid. The important feature of the developed JOL1814 was the delivery of the HA1 antigen to cytosol of peritoneal macrophages. Immunization of chickens with JOL1814 produced significant level of humoral, mucosal, cellular and IL-2, IL-4, IL-17 and IFN-γ cytokine immune response against H5 HA1 and <I>S.</I> Gallinarum antigens in the immunized chickens. Post-challenge, only the JOL1814 immunized chicken showed significantly faster clearance of H5N3 virus in oropharyngeal and cloacal swabs, and 90% survival rate against lethal challenge with a wild type <I>S</I>. Gallinarum. Furthermore, the JOL1814 immunized were differentiated from the H5N3 LPAI virus infected chickens by matrix (M2) gene-specific real-time PCR. In conclusion, the data from the present showed that the JOL1814 can be an effective bivalent vaccine candidate against H5N3 LPAI and fowl typhoid infection in poultry while still offering sero-surveillance property against H5 avian influenza virus.</P>

Orally administered live attenuated <i>Salmonella</i> Typhimurium protects mice against lethal infection with H1N1 influenza virus

Kamble, Nitin Machindra,Hajam, Irshad Ahmed,Lee, John Hwa Elsevier Scientific Pub. Co 2017 Veterinary microbiology Vol.201 No.-

<P><B>Abstract</B></P> <P>Pre-stimulation of toll-like receptors (TLRs) by agonists has been shown to increase protection against influenza virus infection. In this study, we evaluated the protective response generated against influenza A/Puerto Rico/8/1934 (PR8; H1N1) virus by oral and nasal administration of live attenuated <I>Salmonella enterica</I> serovar Typhimurium, JOL911 strain, in mice. Oral and nasal inoculation of JOL911 significantly increased the mRNA copy number of TLR-2, TLR4 and TLR5, and downstream type I interferon (IFN) molecules, IFN-α and IFN-β, both in peripheral blood mononuclear cells (PBMCs) and in lung tissue. Similarly, the mRNA copy number of interferon-inducible genes (ISGs), Mx and ISG15, were significantly increased in both the orally and the nasally inoculated mice. Post PR8 virus lethal challenge, the nasal JOL911 and the PBS control group mice showed significant loss of body weight with 70% and 100% mortality, respectively, compared to only 30% mortality in the oral JOL911 group mice. Post sub-lethal challenge, the significant reduction in PR8 virus copy number in lung tissue was observed in oral [on day 4 and 6 post-challenge (dpc)] and nasal (on 4dpc) than the PBS control group mice. The lethal and sub-lethal challenge showed that the generated stimulated innate resistance (StIR) in JOL911 inoculated mice conferred resistance to acute and initial influenza infection but might not be sufficient to prevent the PR8 virus invasion and replication in the lung. Overall, the present study indicates that oral administration of attenuated <I>S.</I> Typhimurium can pre-stimulate multiple TLR pathways in mice to provide immediate early StIR against a lethal H1N1 virus challenge.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Pre-stimulation of TLR pathways provides stimulated innate resistance (StIR) against influenza virus. </LI> <LI> <I>S.</I> Typhimurium activates multiple TLR pathways, downstream interferons and effector interferon stimulated genes in PBMCs and in lung tissues. </LI> <LI> Oral administration of attenuated <I>S.</I> Typhimurium elicit StIR against PR8 influenza virus. </LI> <LI> The StIR generated in response to <I>S.</I> Typhimurium can protect mice against lethal challenge of PR8 influenza virus. </LI> </UL> </P>

Interaction of a live attenuated<i>Salmonella</i>Gallinarum vaccine candidate with chicken bone marrow-derived dendritic cells

Kamble, Nitin Machindra,Jawale, Chetan Vilas,Lee, John Hwa Informa UK (TaylorFrancis) 2016 Avian pathology Vol.45 No.2

Carpometacarpal Joint Fracture Dislocation of Second to Fifth Finger

Gopal Tukaram Pundkare,Aniket Machindra Patil 대한정형외과학회 2015 Clinics in Orthopedic Surgery Vol.7 No.4

Background: Carpometacarpal joint fracture dislocation of the second to fifth finger is a rare hand injury associated with high energy trauma. Due to severe swelling and overlapping of bones on the radiograph of wrist-hand, dislocations are missed. We reported a series of six patients with rare carpometacarpal joint fracture dislocation treated with open reduction. Methods: We retrospectively studied six cases of carpometacarpal joint fracture dislocation. All patients were treated with open reduction and internal fixation with Kirschner wire. Functional assessment was done with Quick Disabilities of the Arm, Shoulder and Hand score (Quick DASH score) at regular intervals. Results: Average Quick DASH score was improved from 75.76 to 1.9 from 6 weeks to 18 months of duration. Of the six patients, three patients had a Quick DASH score of 0 at the end of 18 months. Conclusions: Careful hand examination and radiographic assessment is necessary to avoid missed diagnosis of carpometacarpal joint fracture dislocation. Early open reduction and internal fixation lead to excellent recovery of hand function.

On nonlinear fractional integro-differential equations with two boundary conditions

SABRI T. M. THABET,MACHINDRA B. DHAKNE 장전수학회 2016 Advanced Studies in Contemporary Mathematics Vol.26 No.3

This paper concerns the existence of solutions of fractional integrodifferential equations with two boundary conditions. Our main analysis based on fractional calculus, Holder inequality and Monch fixed point theorem via Hausdorff’s measure of noncompactness. Example is presented to illustrate the applications of the obtained abstract results.

A live attenuated <i>Salmonella</i> Enteritidis secreting detoxified heat labile toxin enhances mucosal immunity and confers protection against wild-type challenge in chickens

Lalsiamthara, Jonathan,Kamble, Nitin Machindra,Lee, John Hwa BioMed Central 2016 Veterinary research Vol.47 No.-

<P>A live attenuated <I>Salmonella</I> Enteritidis (SE) capable of constitutively secreting detoxified double mutant <I>Escherichia coli</I> heat labile toxin (dmLT) was developed. The biologically adjuvanted strain was generated via transformation of a highly immunogenic SE JOL1087 with a plasmid encoding dmLT gene cassette; the resultant strain was designated JOL1641. A balanced-lethal host-vector system stably maintained the plasmid via auxotrophic host complementation with a plasmid encoded aspartate semialdehyde dehydrogenase (<I>asd</I>) gene. Characterization by western blot assay revealed the dmLT subunit proteins in culture supernatants of JOL1641. For the investigation of adjuvanticity and protective efficacy, chickens were immunized via oral or intramuscular routes with PBS, JOL1087 and JOL1641. Birds immunized with JOL1641 showed significant (<I>P</I> ≤ 0.05) increases in intestinal SIgA production at the 1<SUP>st</SUP> and 2<SUP>nd</SUP> weeks post-immunization via oral and intramuscular routes, respectively. Interestingly, while both strains showed significant splenic protection via intramuscular immunization, JOL1641 outperformed JOL1087 upon oral immunization. Oral immunization of birds with JOL1641 significantly reduced splenic bacterial counts. The reduction in bacterial counts may be correlated with an adjuvant effect of dmLT that increases SIgA secretion in the intestines of immunized birds. The inclusion of detoxified dmLT in the strain did not cause adverse reactions to birds, nor did it extend the period of bacterial fecal shedding. In conclusion, we report here that dmLT could be biologically incorporated in the secretion system of a live attenuated <I>Salmonella</I>-based vaccine, and that this construction is safe and could enhance mucosal immunity, and protect immunized birds against wild-type challenge.</P>

Effect of Preoperative Vitamin D Levels on Functional Performance after Total Knee Arthroplasty

Rajesh Navin Maniar,Aniket Machindra Patil,Adit Rajesh Maniar,Bharat Gangaraju,Jaivardhan Singh 대한정형외과학회 2016 Clinics in Orthopedic Surgery Vol.8 No.2

Background: Low vitamin D levels affects muscle function. Vitamin D and calcium deficiency cause osteomalacic myopathy and poor functional recovery after hip fractures. The relationship of vitamin D and functional performance after total knee arthroplasty (TKA) is not previously reported. Methods: Influence of vitamin D on functional performance before and after TKA was reviewed retrospectively in 120 patients. Of these, 64 had vitamin D deficiency (25-hydroxy vitamin D < 30 ng/mL) preoperatively. All 120 patients received vitamin D oral supplementation postoperatively. Functional parameters including Western Ontario and McMaster Osteoarthritis Index (WOMAC), Short-Form Health Survey questionnaire, and Knee Society Score were assessed pre- and postoperatively at 3 months. Results: Preoperative function was significantly lower in osteoarthritic patients with vitamin D deficiency (WOMAC score; p = 0.040), but at 3 months all functional scores were similar. Conclusions: We concluded that vitamin D deficiency has a negative effect on function in knee osteoarthritic patients. However, postoperative vitamin D supplementation can lead to functional recovery in these patients. Thus, TKA should not be delayed in vitamin D deficient patients; rather supplementation in the postoperative period is preferable to achieve comparable functional outcome at 3 months to patients with vitamin D sufficiency.

Involvement of opioid receptors in antinociceptive activity of semi purified fraction and β-amyrin isolated from Ricinus communis Linn. leaves in mice

Vaishali Murade,Keshav Deshmukh,Rupali Murade,Dipak Raut,Machindra Chavan,Dinesh Hase 경희대학교 융합한의과학연구소 2017 Oriental Pharmacy and Experimental Medicine Vol.17 No.4

The antinociceptive effect of chloroform extract (RCLC), semi-purified fraction (RC2) and isolated pentacyclic triterpenoid β-amyrin (BA) from the leaves of Ricinus communis in acetic acid-induced abdominal writhing, formalin-induced paw licking and hot-plate test was conducted in mice. The GC-MS study revealed that the RC2 showed the dominance of diterpenes, triterpenes, fatty acids, sterol and higher alkanes. No obvious behavioral changes, allergic reactions and mortality were observed in acute toxicity studies. Acute administration of the diazepam (1 mg/kg), RC2 (50 mg/kg) and BA (10 mg/kg) displayed mild sedation in open field test while RCLC (100 mg/kg) showed potent sedative properties. No motor incoordination and neurotoxicity was observed in rota-rod test. RCLC (100 mg/kg), RC2 (25 and 50 mg/kg) and BA (5 and 10 mg/kg) significantly inhibited the number of abdominal writhing induced by acetic acid and formalin-induced paw licking response. The central and peripheral antinociceptive effects for RCLC and RC2 in hot-plate were comparable to morphine. The administration of naloxone reversed the antinociceptive effect of morphine, RCLC and RC2 indicating the involvement of opioid receptors while BA principally showed peripheral antinociceptive effects which may be modulated via peripheral opioid receptors or protein kinase.