Japanese encephalitis virus-induced neuropathology in mouse model infected through the conjunctival route

Background & objectives: Mouse is a preferred animal model for studying pathogenesis of Japanese encephalitis virus (JEV) infections, and different routes of inoculation have been tried. Some neurotropic viruses can reach the brain following infection through ocular route. This study was undertaken to establish JEV-induced clinical disease in mouse model through conjunctival route and document the neuropathological effects. Methods: Ten two-week old Swiss albino mice were inoculated with 5 ?l Vero cell cultured virus containing 104.7 TCID50 JEV through conjunctival route. Clinical signs of mice were observed twice daily. After necropsy examination, different organs including eyes and olfactory bulbs were collected for histopathological examination, quantification of viral copy number and antigen by real-time TaqMan assay and immunohistochemistry, respectively. Results: Infected mice showed characteristic clinical signs of JE by 4 days post-infection (dpi). Histopathological lesions in brain included perivascular cuffing by mononuclear cells, focal gliosis, necrosis of neurons and neuronophagia and astrocytosis in the cerebrum, cerebellum and the brainstem. JEV viral load was highest in the brain followed by intestine, heart, liver, spleen, lung and kidney. JEV antigen was detected in the bipolar and ganglion cells of the retina and in the mitral cells and periglomerular cells of olfactory bulb and other parts of the brain. Interpretation & conclusions: JEV infection in mice through conjunctival route produced characteristic clinical signs of the disease and neuropathological lesions. Demonstration of JEV antigen in association with neuropathological lesions in the central nervous system and neuronal cells of the eye showed that conjunctival route could be an effective alternate route for virus invasion into the brain. These findings have biosafety implications for researchers, veterinary practitioners and pig farmers.

Bony Impressions on Caput and Neck of Human Femora in Indian Population

Idiopathic osteoarthritis of the hip has been currently attributed by researchers to the repetitive microtrauma caused to the femoral neck by its impingement against the acetabular rim. This impingement occurs as a result of abnormal morphological traits like Allen’s fossa, Poirier’s Facet, and Posterior Cervical Imprint that appear on the proximal end of femur especially on the neck. Hence, this study was undertaken to investigate the incidence of these traits in the Indian population and to find a correlation between the occurrence of the trait and the side and sex of the bone. The study was conducted on 152 adult dried femora in the Department of Anatomy, Maulana Azad Medical College, New Delhi, India. The positive findings were photographed. The significance of the correlation was found out using Chi-square test. Allen’s Fossa was found to occur in 71.1% of the total bones assessed Poirier’s Facet in 31.6% and Posterior Cervical Imprint in 19.7%. Out of these, a significant side association was found for the Poirier’s Facet with a predominance of the left side. All the traits were found to be more in males with a significant sex variation for Poirier’s Facet. Each trait demonstrated a significant side and gender dimorphism.

Clinical Pulmonary Infection Score to Diagnose Ventilator-associated Pneumonia in Children.

Background: There is a need to validate and suggest easy clinical method for diagnosis of ventilator-associated pneumonia (VAP) in developing countries. Objectives: To validate the use of simplified Clinical Pulmonary Infection Score (CPIS) for the diagnosis of VAP. Design: Prospective study. Setting: Pediatric intensive care unit of a tertiary care teaching hospital. Subjects: 30 children receiving mechanical ventilation for more than 48 hours and with simplified CPIS6. Methods: All patients underwent flexible bronchoscopy to obtain bronchoalveolar lavage which was analyzed quantitatively. Colony count 104cfu/mL was considered reference standard for definite VAP. Results: Of the five variables used for simplified CPIS, only patient’s temperature (P=0.013) and PaO2/ FiO2 ratio were significant (P<0.001) to differentiate the presence of definite VAP. Patients with definite VAP (BAL colony count 104cfu/mL) had CPIS of 8.4 while in no definite VAP group it was 6.4 (P= 0.007). CPIS of 8 was found to have sensitivity of 80%, specificity 80%, PPV 86.9%, NPV 70.5% and accuracy 80%. The area under Receiver operating characteristic curve of CPIS against reference standard was 0.81± 0.069 (P=0.001). Conclusion: Simplified CPIS is useful in patients on mechanical ventilation to diagnose ventilator- associated pneumonia.

Treatment of enteric fever in children on the basis of current trends of antimicrobial susceptibility of Salmonella enterica serovar typhi and paratyphi A.

PURPOSE: Recent reports indicate decreased susceptibility of S. typhi to fluoroquinolones, especially ciprofloxacin. Chloramphenicol has been suggested as first line therapy of enteric fever in many studies. This is a prospective study that describes the trends of antimicrobial susceptibility of S. typhi and S. paratyphi A causing bacteraemia in children and reports therapeutic failure to ciprofloxacin and evaluates the possible use of chloramphenicol, ampicillin, ciprofloxacin and third generation cephalosporins as first line therapy in the treatment of enteric fever in children. METHODS: The present study was conducted from April 2004 to March 2005 in a superspeciality children hospital at New Delhi. A total of 56 S. typhi and five S. paratyphi A isolates were obtained among the 673 blood cultures performed. Antimicrobial testing was done using disk diffusion technique (NCCLS method) for 13 antimicrobials and MICs were calculated for ampicillin, ciprofloxacin, chloramphenicol and cefotaxime. Analysis of data was done using WHONET software. RESULTS: All 56 isolates of S. typhi were sensitive to amoxycillin+clavulanate, gentamicin, cefixime, cefotaxime and ceftazidime. Multidrug resistance (MDR, resistance to three drugs) was seen in 22 cases (39%) and resistance to five drugs was seen in 12 cases (21%). Only two isolates were resistant to chloramphenicol (3%). MIC 90 for ampicillin, chloramphenicol, ciprofloxacin and cefotaxime were 1.0 microg/ml, 4.0 microg/ml, 64 microg/ml and 0.125 microg/ml respectively. All S. paratyphi A isolates were sensitive to ampicillin and chloramphenicol and resistant to nalidixic acid. MIC distribution data for chloramphenicol revealed elevated MIC but still in susceptible range. CONCLUSIONS: There is an urgent need for further clinical studies to evaluate response to chloramphenicol in such cases. Antimicrobial susceptibility data and MIC distribution favour use of ampicillin as a drug of choice for the treatment of enteric fever. Third generation cephalosporins are also useful but their use should be restricted for complicated cases.

Post kala-azar dermal leishmaniasis (PKDL): a first case report from Nepal.

Post kala-azar dermal leishmaniasis (PKDL) is a non-ulcerative lesion of the skin caused by Leishmania donovani, which is usually seen after completion of treatment of the kala-azar. The condition is yet to be reported from Nepal. We document and report for the first time a case of PKDL in Nepal.

Post-kala-azar dermal leishmaniasis (PKDL): the first case report from Nepal.

Post-kala-azar dermal leishmaniasis (PKDL) is condition characterized by non-ulcerative lesions of the skin caused by Leishmania donovani that is usually seen after the completion of treatment of kala-azar (visceral leishmaniasis). We document the first case report of PKDL in Nepal.

Cutaneous leishmaniasis in Nepal.

We report an imported case of cutaneous leishmaniasis in a 30 year old adult male from Nepal caused by Leishmania tropica. This case from Dharan is the first such report of imported cutaneous leishmaniasis in Nepal.