Incidence of Japanese Encephalitis and Acute Encephalitis Syndrome Hospitalizations in the Medium-Endemic Region in Central India.

BACKGROUND: We estimated the incidence of Japanese encephalitis (JE) and acute encephalitis syndrome (AES) following routine immunization with the live-attenuated SA 14-14-2 JE vaccine. METHODS: We implemented enhanced surveillance of AES and JE hospitalizations in endemic districts in Maharashtra and Telangana States during 2015-2016 and 2018-2020. We estimated incidence and compared differences in the incidence of JE and AES between two states, and vaccinated and unvaccinated districts during two study periods. We also considered secondary data from public health services to understand long-term trends from 2007 to 2020. RESULTS: The annual AES incidence rate of 2.25 cases per 100,000 children in Maharashtra during 2018-2020 was significantly lower than 3.36 cases per 100,000 children during 2015-2016. The six JE-vaccinated districts in Maharashtra had significantly lower incidence rates during 2018-2020 (2.03, 95% CI 1.73-2.37) than in 2015-16 (3.26, 2.86-3.70). In addition, the incidence of both JE and AES in two unvaccinated districts was higher than in the vaccinated districts in Maharashtra. Telangana had a lower incidence of both JE and AES than Maharashtra. The AES incidence rate of 0.95 (0.77-1.17) during 2018-2020 in Telangana was significantly lower than 1.67 (1.41-1.97) during 2015-2016. CONCLUSIONS: The annual incidence rate of Japanese encephalitis was < 1 case per 100,000 children. It indicated accelerated control of Japanese encephalitis after routine immunization. However, the annual incidence of acute encephalitis syndrome was still > 1 case per 100,000 children. It highlights the need for improving surveillance and evaluating the impacts of vaccination.

Effectiveness of Japanese encephalitis vaccination among children in central India.

Japanese encephalitis (JE) disease among children continues in central India despite vaccination implemented in the routine immunization program. Therefore, we planned to estimate the JE vaccination effectiveness among children by undertaking a 1:2 individually-matched population-based case-control study from August 2018 to October 2020. The laboratory-confirmed JE cases aged 1-15 years were enrolled along with neighborhood controls without fever and encephalitis matched on the residence area, age and sex. The JE vaccination history was enquired from parents and verified independently from the vaccination cards available at home and records at health facilities. We enrolled 35 JE cases and 70 matched controls. The vaccination effectiveness of 86.7% (95% confidence interval [CI]: 30.8-94.7) was estimated on the per-protocol analysis of 31 case-control sets. The screening method provided an effectiveness of 89.5% (CI: 78.9-94.7) on using the population vaccination coverage of 90% reported earlier in the same area. In conclusion, JE vaccination offered a moderate level of protection among children in JE medium-endemic central India, similar to reports from high-endemic areas in India. The operational aspects of vaccination program implementation need to be evaluated to assess the impact of vaccination on the disease burden of JE in medium-endemic regions of India.

Evaluation of USP22 and Ki-67 expression in oral squamous cell carcinoma: An immunohistochemical study.

Background and Aim: USP22 is a positive regulator in tumor growth, its depletion leads to cell cycle arrest at G1 phase. USP22 over expression was positively correlated with proteins involved in proliferation and negatively correlated with tumor suppressor protein tumor supprn. Ki-67 expression is associated with USP22 over expression in oral squamous cell carcinoma (OSCC) and also in cervical and prostate cancers. The aim of this study is to evaluate the expression of USP22 and Ki-67 in OSCC by using an immunohistochemical staining procedure. Materials and Methods: Immunohistochemistry was used to determine the expression of USP22 protein in 50 archival tissue blocks of histopathologically diagnosed OSCC and 15 normal oral mucosa tissue blocks. The histopathological correlation of USP22 with Ki-67 was done. Results: Expression of USP22 and Ki-67 was seen in the nuclei of epithelial cells. Statistical analysis of the mean expression of USP22 in OSCC and normal tissue showed a significant difference (P = 0.000000119). A significant difference was also observed in Ki-67 between OSCC and normal tissue (P = 0.00000086). Correlation test showed a weak correlation (R = 0.19) between USP22 and Ki-67 expression of group 1. Similarly, a weak correlation (R = 0.51) was observed in group 2. Conclusion: A statistically significant difference in the expression of USP22 and Ki-67 was observed between normal mucosa and OSCC. It can be used in early diagnosis of OSCC but its use as a prognostic indicator is questionable and should be exemplified with a larger study sample.

Infectious causes of acute encephalitis syndrome hospitalizations in Central India, 2018-20.

BACKGROUND: We enhanced surveillance of hospitalizations of all ages for acute encephalitis syndrome (AES) along with infectious aetiologies, including the Japanese encephalitis virus (JEV). METHODS: From October 2018 to September 2020, we screened neurological patients for AES in all age groups in Maharashtra and Telangana States. AES cases were enrolled at study hospitals along with other referrals and sampled with cerebrospinal fluid, acute and convalescent sera. We tested specimens for non-viral aetiologies viz. leptospirosis, typhoid, scrub typhus, malaria and acute bacterial meningitis, along with viruses - JEV, Dengue virus (DENV), Chikungunya virus (CHIKV), Chandipura virus (CHPV) and Herpes simplex virus (HSV). RESULTS: Among 4977 neurological hospitalizations at three study site hospitals over two years period, 857 (17.2%) were AES. However, only 287 (33.5%) AES cases were eligible. Among 278 (96.9%) enrolled AES cases, infectious aetiologies were identified in 115 (41.4%) cases, including non-viral in 17 (6.1%) cases - leptospirosis (8), scrub-typhus (3) and typhoid (6); and viral in 98 (35.3%) cases - JEV (58, 20.9%), HSV (22, 7.9%), DENV (15, 5.4%) and CHPV (3, 1.1%). JEV confirmation was significantly higher in enrolled cases than referred cases (10.2%) (p < 0.05). However, the contribution of JEV in AES cases was similar in both children and adults. JE was reported year-round and from adjacent non-endemic districts. CONCLUSIONS: The Japanese encephalitis virus continues to be the leading cause of acute encephalitis syndrome in central India despite vaccination among children. Surveillance needs to be strengthened along with advanced diagnostic testing for assessing the impact of vaccination.

Neonatal Joubert Syndrome With Renal Involvement and Respiratory Distress.

Joubert syndrome (JS) is a rare autosomal recessive neurodevelopmental disorder with characteristic clinical presentation of hyperpnea-apnea spells, hypotonia, dysmorphic facies, and nystagmus and imaging features of molar tooth sign and cerebellar vermian hypoplasia-dysplasia. Early diagnosis is needed for timely management and favorable outcome. We present a case of neonatal JS with renal involvement presenting with respiratory distress and highlight the characteristic clinical and imaging findings. On examination, the baby had low set ears, a large protruding tongue, hypertelorism, and a depressed nasal bridge. Ultrasonography (USG) abdomen showed echogenic kidneys with cortical and medullary cysts. Magnetic Resonance Imaging (MRI) brain showed classical molar tooth sign, vermian hypoplasia-dysplasia, and thinning of the corpus callosum.

Childhood encephalitis hospitalizations associated with virus agents in medium-endemic states in India.

BACKGROUND: Japanese encephalitis (JE) is the leading cause of childhood acute encephalitis syndrome (AES) in India. We enhanced the AES surveillance in sentinel hospitals to determine trends and virus etiologies in central India. METHODS: The neurological hospitalizations among children ≤15 years were tracked by using the AES case definition implemented by the national program. Acute and convalescent sera along with cerebrospinal fluid (CSF) specimens were collected and tested at the strengthened site hospital laboratories for anti-JE, anti-Dengue and anti-Chikungunya virus by IgM ELISA; along with Chandipura virus RT-PCR. Herpes simplex and enterovirus testing was undertaken at the reference laboratory. RESULTS: Among 1619 pediatric neurological hospitalizations reported during 2015-16, AES case definition was fulfilled in 332 (20.5%) cases. After excluding 52 non-AES cases, 280 AES cases resident from study districts were considered eligible for study. The treating physicians diagnosed non-viral causes in 90 cases, therefore 190 (67.9%) of 280 AES cases were suspected with viral etiologies. We enrolled 140 (73.7%) of 190 eligible AES cases. Viral etiologies were confirmed in 31 (22.1%) of 140 enrolled AES cases. JE (n = 22) was the leading cause. Additional non-JE viral agents included Chikungunya (5), Dengue (2) and Chandipura (2). However, only 21 (9.4%) of 222 additional AES cases referred from peripheral hospitals were confirmed as JE. CONCLUSIONS: Japanese encephalitis virus continues to be the leading cause of childhood acute encephalitis syndrome in central India despite vaccination program. Surveillance needs to be intensified for assessing the true disease burden of Japanese encephalitis following vaccination program implementation.

Collagen during odontogenesis and in ameloblastoma: A polarizing microscopic study.

BACKGROUND: Odontogenesis is a highly coordinated and complex process which depends on cell-cell interactions that result in initiation and generation of tooth. Tissue remnants of developing tooth can form odontogenic tumors possibly, reflecting different developmental stages in tooth formation. In both odontogenesis and odontogenic tumors, stroma plays a prominent role in maintaining epithelial tissues with continuous molecular interactions. As the collagen forms an integral part of connective tissue stroma, in the present study, polarization colors and thickness of the collagen fibers were assessed in both tooth germ papillae and ameloblastoma using picrosirius red (PSR) stain. MATERIALS AND METHODS: Collagen fibers in 20 cases of ameloblastoma and 10 tooth germs from the human fetus were evaluated with PSR stain and examined under polarizing microscopy. RESULTS: Polarization colors of red-colored collagen fibers with greater diameter were more in ameloblastoma when compared to tooth germ papillae in which green-colored collagen fibers with smaller diameter being more. CONCLUSION: The absence of hard tissue formation in ameloblastoma might be due to the presence of significantly more number and greater thickness of red-colored collagen fibers. Thus, the nature of collagen fibers can predict the nature in terms of biologic behavior and prognosis.

Indian Academy of Pediatrics (IAP) Advisory Committee on Vaccines and Immunization Practices (ACVIP) Recommended Immunization Schedule (2018-19) and Update on Immunization for Children Aged 0 Through 18 Years.

JUSTIFICATION: There is a need to revise/review recommendations regarding existing vaccines in view of current developments in vaccinology. PROCESS: Advisory Committee on Vaccines and Immunization Practices (ACVIP) of Indian Academy of Pediatrics (IAP) reviewed the new evidence, had two meetings, and representatives of few vaccine manufacturers also presented their data. The recommendations were finalized unanimously. OBJECTIVES: To revise and review the IAP recommendations for 2018-19 and issue recommendations on existing and certain new vaccines. RECOMMENDATIONS: The major changes in the IAP 2018-19 Immunization Timetable include administration of hepatitis B vaccine within 24 hours of age, acceptance of four doses of hepatitis B vaccine if a combination pentavalent or hexavalent vaccine is used, administration of DTwP or DTaP in the primary series, and complete replacement of oral polio vaccine (OPV) by injectable polio vaccine (IPV) as early as possible. In case IPV is not available or feasible, the child should be offered three doses of bivalent OPV. In such cases, the child should be advised to receive two fractional doses of IPV at a Government facility at 6 and 14 weeks or at least one dose of intramuscular IPV, either standalone or as a combination, at 14 weeks. The first dose of monovalent Rotavirus vaccine (RV1) can be administered at 6 weeks and the second at 10 weeks of age in a two-dose schedule. Any of the available rotavirus vaccine may be administered. Inactivated influenza vaccine (either trivalent or quadrivalent) is recommended annually to all children between 6 months to 5 years of age. Measles-containing vaccine (MMR/MR) should be administered after 9 months of age. Additional dose of MR vaccine may be administered during MR campaign for children 9 months to 15 years, irrespective of previous vaccination status. Single dose of Typhoid conjugate vaccine (TCV) is recommended from the age of 6 months and beyond, and can be administered with MMR vaccine if administered at 9 months. Four-dose schedule of anti-rabies vaccine for Post Exposure Prophylaxis as recommended by World Health Organization in 2018, is endorsed, and monoclonal rabies antibody can be administered as an alternative to Rabies immunoglobulin for post-exposure prophylaxis.

Evaluation of cytomorphometric changes in tobacco users and diagnosed oral squamous cell carcinoma individuals.

AIMS: To determine the cellular and nuclear area of keratinocytes in smears obtained from the oral mucosa of tobacco users, those with oral squamous cell carcinoma (OSCC), and from normal healthy persons and resolve if any significant difference exists in these three groups. MATERIALS AND METHODS: The study group comprised 100 subjects 20 controls, (40 OSCC patients-20 from lesional sites and 20 from nonlesional sites, 20 tobacco smokers and 20 tobacco chewers) in the age group of 25-75 years. Oral mucosal smears obtained by using a cytobrush were stained with Papanicolaou (PAP) stain and using 20X objective in trinocular Olympus model BX53 with Jenoptik scientific grade-dedicated microphotographic camera images were taken. With ProgRes version 8.0 image analysis software, 20 cells with defined borders were evaluated from each slide. Finally, one-way analysis of variance (ANOVA) was used to compare the above parameters in the studied groups. STATISTICAL ANALYSIS USED: Minitab and Excel software were used to analyze the data. One-way ANOVA was used to compare the above parameters in the studied groups. RESULTS: The mean value of the cell area for groups I, II, III, IV, and V were 2838 ± 275.2, 2762.1 ± 511.4, 2861.9 ± 512.9, 2643.8 ± 333.3, and 3064.3 ± 362.7, respectively, the nuclear area (NA) was 83.88 ± 9.86, 106.19 ± 13.45, 95.11 ± 14.24, 85.55 ± 21.11, and 80.83 ± 13.45, respectively, and nuclear-to-cellular (N:C) ratio was 0.0297, 0.03924, 0.0337, 0.03257, and 0.02678, respectively. CONCLUSIONS: Thus, our study elucidates that cytomorphology gauges the effect of tobacco on the oral mucosa and possibly establishes a link between premalignant and malignant transformations even before a lesion is visibly noted.

Cytological grading: An alternative to histological grading in oral squamous cell carcinoma.

BACKGROUND: Micronuclei (MN) in oral exfoliative cells have been shown to indicate a disparaging change in genetic information of the cell. Recent studies showed correlation between the frequency of MN and severity of this damage. Grading of lesions can be used to determine the austerity of this damage. Aims: The aim of this study is to compare the MN frequency in oral exfoliated cells of normal and oral squamous cell carcinoma (OSCC) individuals and to cytologically grade the frequency of MN in cytological smears and to correlate it with histological grading. The objective is to ascertain whether MN frequency in oral exfoliated cells can be a parameter for grading of OSCC. SETTINGS AND DESIGN: The study group comprises of 40 subjects (20 controls and 20 OSCC patients) in the age group of 45-85 years. MATERIALS AND METHODS: The cytosmear was obtained from each group and stained with Papanicolaou (PAP) stain. Twenty cells from each slide were counted for MN and cytological grade of OSCC was assigned based on the average frequency of MN. Cytological grade was correlated with histological grading and the data were recorded. Student's t-test and Spearman's correlation were used for the analysis of the data. RESULTS: Average frequency of MN was 2.5 times higher in OSCC patients when compared to that in controls and the difference was found to be highly significant. Sixty percent correlation was found between cytological grade and histological grade of OSCC and the difference between them was not significant. CONCLUSIONS: Cytological grading can be used in grading OSCC, and MN insinuates genotoxic damage occurring in the epithelial cells.

Certainty of S100 from Physiology to Pathology.

S100 exists in wide variety of tissues and cell types, originally isolated from brain tissue and they are of low molecular weight proteins. S100 is evenly distributed in cytoplasm and also in nucleoplasm and is involved in both intercellular and extracellular functions. S100 protein is generally expressed in normal and also in pathological conditions. In current review, we discuss: a) update nomenclature of the various S100 proteins, b) expression of S100 in oral diseases (different soft tissue tumors, odontogenic cyst and tumor) for diagnostic value and also to know their histogenesis, c) role of S100 and RAGE receptor in oral squamous cell carcinoma.

Rare case report of Traumatic neuroma of anterior superior alveolar nerve associated with high frenal attachment.

We present an incredible case of traumatic neuroma (TN) in the anterior superior alveolar nerve leading to the swelling in the upper labial mucosa. This paper attempts to highlight the rarity of site of occurrence of this lesion and reports the first case of TN of anterior superior alveolar nerve.

A qualitative and quantitative analysis of AgNORs in keratocystic odontogenic tumor, unicystic ameloblastoma and multicystic ameloblastoma.

AIMS: A comparative evaluation of proliferation activity in unicystic ameloblastoma (UA), multicystic ameloblastoma (MA) and keratocystic odontogenic tumor (KCOT) using silver staining technique. SETTINGS AND DESIGN: In the present study 21 histopathologically confirmed paraffin blocks,7 each of UA, MA and KCOT were selected and stained with silver nitrate. MATERIALS AND METHODS: For quantitative analysis, 100 cells were counted at 1000x magnification for AgNORs and the mean value was calculated. Qualitative analysis of AgNORs included normal (oval shaped) and abnormal groups (bean shaped) in the lesion. STATISTICAL ANALYSIS: The statistical analysis of data was done by a specialist statistician using two way ANOVA and multiple comparisons with Tukey's test in advanced excel. RESULTS: The AgNOR count was more in KCOT when compared to MA and UA with the pattern of distribution of AgNORs more in basal than in the parabasal layer in KCOT. The qualitative analysis showed small to large oval AgNOR's in KCOT and few clusters in MA whereas in UA irregular clusters were seen. CONCLUSION: This concludes the expediency of AgNOR staining in reflecting the high proliferation rate and a more aggressive behavior of KCOT in comparison to MA and UA which signifies requirement of a more hostile surgical approach in KCOT to avoid recurrences following different treatment modalities.

Efficacy of 1.5% dish washing solution and 95% lemon water in substituting perilous xylene as a deparaffinizing agent for routine h and e staining procedure: a short study.

Aim. To assess the efficacy of dish washing solution and diluted lemon water in deparaffinizing sections during conventional hematoxylin and eosin staining technique. Objective. The objective is to utilize eco-friendly economical substitute for xylene. Materials and Methods. Using twenty paraffin embedded tissue blocks, three sections each were prepared. One section was stained with conventional H and E method (Group A) and the other two sections with xylene-free (XF) H and E (Groups B and C). Staining characteristics were compared with xylene and scoring was given. Total score of 3-5 was regarded as adequate for diagnosis and less than that inadequate for diagnosis. Statistical Analysis. Chi-square test, Kruskal Wallis ANOVA test, and Mann-Whitney U test were used. Results. Adequacy of nuclear staining, crispness, and staining for diagnosis were greater in both Groups A and C (100%) than Group B (95%). Adequacy of cytoplasmic staining was similar in all the three groups (100%). Group B showed comparatively superior uniform staining and less retention of wax. Conclusion. Dish washing solution or diluted lemon water can be replaced for xylene as deparaffinizing agent in hematoxylin and eosin procedure.

Mucormycosis: an atrocious mate of patients with diabetes.

Mucormycosis also called zygomycosis is a rare infection caused by saprophytic aerobic fungus that belongs to a group of fungi called Mucoromycotina in the order Mucorales. Earlier these fungi were called Zygomycota but this scientific name has recently been changed. Mucormycosis can result in an acute, rapidly advancing and occasionally fatal disease caused by different fungi typically found in the soil and in association with decaying organic matter such as leaves, compost piles or rotten wood. These fungal infections are relatively infrequent; however, they occur in individuals who are debilitated in some major way and occasionally in groups of people with multiple penetrating injuries that are contaminated with soil and water from the environment. Mucormycosis is not contagious and does not spread from person to person.