Effectiveness of 13-valent pneumococcal conjugate vaccine on radiological primary end-point pneumonia among cases of severe community acquired pneumonia in children: A prospective multi-site hospital-based test-negative study in Northern India.

INTRODUCTION: Community acquired pneumonia (CAP) is a leading cause of under-five mortality in India and Streptococcus pneumoniae is the main bacterial pathogen for it. Pneumococcal Conjugate Vaccine 13 (PCV13) has been introduced in a phased manner, in the national immunization program of India since 2017/2018. The primary objective of this study was to evaluate the effectiveness of PCV13 on chest radiograph (CXR)-confirmed pneumonia, in children hospitalized with WHO-defined severe CAP. METHODS: This prospective, multi-site test-negative study was conducted in a hospital-network situated in three districts of Northern India where PCV13 had been introduced. Children aged 2-23 months, hospitalized with severe CAP and with interpretable CXR were included after parental consent. Clinical data was extracted from hospital records. CXRs were interpreted by a panel of three independent blinded trained radiologists. Exposure to PCV13 was defined as ≥2 doses of PCV13 in children aged ≤ 12 months and ≥ 1 dose(s) in children > 12 months of age. Our outcome measures were CXR finding of primary endpoint pneumonia with or without other infiltrates (PEP±OI); vaccine effectiveness (VE) and hospital mortality. RESULTS: From 1st June 2017-30th April 2021, among 2711 children included, 678 (25.0%) were exposed to PCV1. CXR positive for PEP±OI on CXR was found in 579 (21.4%), of which 103 (17.8%) were exposed to PCV. Adjusted odds ratio (AOR) for PEP±OI among the exposed group was 0.69 (95% CI, 0.54-0.89, p = 0.004). Adjusted VE was 31.0% (95% CI: 11.0-44.0) for PEP±OI. AOR for hospital mortality with PEP±OI was 2.65 (95% CI: 1.27-5.53, p = 0.01). CONCLUSION: In severe CAP, children exposed to PCV13 had significantly reduced odds of having PEP±OI. Since PEP±OI had increased odds of hospital mortality due to CAP, countrywide coverage with PCV13 is an essential priority.

Diffusion Tensor and Dynamic Contrast-Enhanced Magnetic Resonance Imaging Correlate with Molecular Markers of Inflammation in the Synovium.

Objectives: It is difficult to capture the severity of synovial inflammation on imaging. Herein we hypothesize that diffusion tensor imaging (DTI) derived metrics may delineate the aggregation of the inflammatory cells and expression of inflammatory cytokines and dynamic contrast-enhanced (DCE) imaging may provide information regarding vascularity in the inflamed synovium. Patients and methods: Patients with knee arthritis (>3-months duration) underwent conventional (T2-weighted fast spin echo and spin echo T1-weighted images) as well as DTI and DCE MRI and thereafter arthroscopic guided synovial biopsy. DCE and DTI metrics were extracted from the masks of the segments of the inflamed synovium which enhanced on post-contrast T1-weighted MRI. These metrics were correlated with immunohistochemistry (IHC) parameters of inflammation on synovium. Statistical analysis: Pearson's correlation was performed to study the relationship between DTI- and DCE-derived metrics, IHC parameters, and post-contrast signal intensity. Linear regression model was used to predict the values of IHC parameters using various DTI and DCE derived metrics as predictors. Results: There were 80 patients (52 male) with mean age 39.78 years and mean disease duration 19.82 months. Nineteen patients had tuberculosis and the rest had chronic undifferentiated monoarthritis (n = 31), undifferentiated spondyloarthropathy (n = 14), rheumatoid arthritis (n = 6), osteoarthritis (n = 4), reactive arthritis (n = 3), ankylosing spondylitis (n = 2), and juvenile idiopathic arthritis (n = 1). Fractional anisotropy (FA), a metric of DTI, had significant correlation with number of immune cells (r = 0.87, p < 0.01) infiltrating into the synovium and cytokines (IL-1ß, r = 0.55, p < 0.01; TNF-α, r = 0.42, p < 0.01) in all patients and also in each group of patients and adhesion molecule expressed on these cells in all patients (CD54, r = 0.51, p < 0.01). DCE parameters significantly correlated with CD34 (blood flow, r = 0.78, p < 0.01; blood volume, r = 0.76, p < 0.01) in each group of patients, a marker of neo-angiogenesis. FA was the best predictor of infiltrating inflammatory cells, adhesion molecule and proinflammatory cytokines. Amongst the DCE parameters, blood volume, was best predictor of CD34. Conclusion: DTI and DCE metrics capture cellular and molecular markers of synovial inflammation in patients with chronic inflammatory arthritis.

Epidemiology of Hypoxic Community-Acquired Pneumonia in Children Under 5 Years of Age: An Observational Study in Northern India.

BACKGROUND: Community-acquired pneumonia (CAP) is the leading cause of under-five mortality in India. An increased risk of mortality has been reported in cases of hypoxic pneumonia. METHODS: The primary objective of this study was to assess the proportion of children aged 2-59 months, hospitalized with hypoxic CAP, as well as socio-demographic, clinical, and radiological features associated with it. The secondary objective was to determine the risk of mortality among hospitalized cases of hypoxic CAP. This prospective, observational study was conducted in four districts of Northern India, between January 2015 and April 2021. A hospital-based surveillance network was established. Inclusion criteria were as follows: (a) child between 2 and 59 months, (b) hospitalization with symptoms of WHO-defined CAP, (c) resident of project district, (d) illness of <14 days, and (e) child had neither been hospitalized for this illness nor recruited previously. Children whose chest x-rays (CXRs) were either unavailable/un-interpretable and those that received any dose of pneumococcal conjugate vaccine-13 were excluded. Hypoxic pneumonia was defined as oxygen saturation <90% on pulse oximetry or requiring oxygen supplementation during hospital stay. RESULTS: During the study period, 71.9% (7,196/10,006) children of severe pneumonia were eligible for inclusion, of whom 35.9% (2,580/7,196) were having hypoxic pneumonia. Female gender and use of biomass fuel for cooking increased the odds of hypoxic CAP. Clinical factors like wheezing, pallor, tachypnea, low pulse volume, presence of comorbidity, general danger signs, severe malnutrition, and radiological finding of primary end-point pneumonia ± other infiltrates (PEP±OI) also increased the odds of hypoxic CAP in a conditional logistic regression model. Adjusted odds ratio for mortality with hypoxia was 2.36 (95% CI: 1.42-3.92). CONCLUSION: Almost one-third of cases hospitalized with severe CAP had hypoxia, which increased chances of mortality. Besides known danger signs, certain newer clinical signs such as pallor and wheezing as well as PEP+OI were associated with hypoxic CAP. Therefore, objective assessment of oxygen saturation must be done by pulse oximetry in all cases of CAP at the time of diagnosis.

Chest radiograph findings in children aged 2-59 months hospitalised with community-acquired pneumonia, prior to the introduction of pneumococcal conjugate vaccine in India: a prospective multisite observational study.

OBJECTIVES: The current study was a hospital-based surveillance of cases hospitalised with WHO-defined community-acquired pneumonia in children aged 2-59 months, to assess the radiological abnormalities in chest X-rays and to identify the demographic and clinical correlates of specific radiological abnormalities, in residents of prespecified districts of Uttar Pradesh and Bihar, India. DESIGN: Prospective, active, hospital-based surveillance. SETTING: Multisite study conducted in a network of 117 secondary/tertiary care hospitals in four districts of Uttar Pradesh and Bihar, India. PARTICIPANTS: Included were children aged 2-59 months, hospitalised with community-acquired pneumonia, residing in the project district, with duration of illness <14 days and who had not been hospitalised elsewhere for this episode nor had been recruited previously. MAIN OUTCOME MEASURE: Concordant radiological abnormalities in the chest X-rays. RESULTS: From January 2015 to April 2017, 3214 cases were recruited and in 99.40% (3195/3214) chest X-rays were available, among which 88.54% (2829/3195) were interpretable. Relevant radiological abnormalities were found in 34.53% (977/2829, 95% CI 32.78 to 36.28). These were primary end point pneumonia alone or with other infiltrates in 22.44% (635/2829, 95% CI 20.90% to 23.98%) and other infiltrates in 12.09% (342/2829; 95% CI 10.88% to 13.29%). There was a statistically significant interdistrict variation in radiological abnormalities. Statistically significantly higher proportion of abnormal chest X-rays were found in girls, those with weight-for-age z-score ≤-3SD, longer duration of fever, pallor and with exposure to biomass fuel. CONCLUSIONS: Among hospitalised cases of community-acquired pneumonia, almost one-third children had abnormal chest radiographs, which were higher in females, malnourished children and those with longer illnesses; and an intra-district variation was observed.

Effectiveness of various communication strategies for improving childhood pneumonia case management: a community based behavioral open labeled trial in rural Lucknow, Uttar Pradesh, India.

BACKGROUND: Community acquired pneumonia is responsible for 16% of under 5 mortality in India, probably due to delayed recognition and qualified care seeking. Therefore these deaths could possibly be averted by creating community awareness and promoting care seeking from qualified physicians in the government system. The objective of study was to assess the effectiveness of facility-based and village-based behavior change communication interventions delivered to community using validated information, education and communication materials, along with infrastructural strengthening of health facilities, for change in care seeking from government system for community acquired pneumonia in rural Lucknow, India. METHOD: Community based open labeled behavioral trial in 2 by 2 factorial design was conducted in eight rural blocks of Lucknow, northern India. Trained community health workers conducted Pneumonia Awareness Sessions once a month for the care givers of children using validated information, education and communication materials either at the villages or at government health facilities. Prior infrastructural strengthening of public health facilities was done to provide optimal care to cases. Pre packed pneumonia drug kits were provided which had amoxicillin, paracetamol and an instruction card on their use as well as pictorial representation of danger signs of pneumonia. RESULTS: Study lasted from October 2015 to September 2018. Adherence to conduct of facility-based intervention was 93.0% (279/300) and to village-based intervention was 73.4% (7638/10410). In village-based intervention there was 79.3% (p < 0.0001) increase from a baseline of 3.3% (14/420) and facility-based intervention 68.9% (p = 0.02) increase from a baseline of 5.35% (21/392) in cases of possible pneumonia treated at government health facilities. CONCLUSION: Conduct of structured pneumonia awareness session using validated information, education and communication material at village level with infrastructural strengthening resulted in improved qualified care seeking from government facilities for community acquired pneumonia. TRIAL REGISTRATION: AEARCTR-0003137, retrospectively registered on 10/July/2018.

Selection of appropriate statistical methods for data analysis.

In biostatistics, for each of the specific situation, statistical methods are available for analysis and interpretation of the data. To select the appropriate statistical method, one need to know the assumption and conditions of the statistical methods, so that proper statistical method can be selected for data analysis. Two main statistical methods are used in data analysis: descriptive statistics, which summarizes data using indexes such as mean and median and another is inferential statistics, which draw conclusions from data using statistical tests such as student's t-test. Selection of appropriate statistical method depends on the following three things: Aim and objective of the study, Type and distribution of the data used, and Nature of the observations (paired/unpaired). All type of statistical methods that are used to compare the means are called parametric while statistical methods used to compare other than means (ex-median/mean ranks/proportions) are called nonparametric methods. In the present article, we have discussed the parametric and non-parametric methods, their assumptions, and how to select appropriate statistical methods for analysis and interpretation of the biomedical data.

Incidence of community acquired pneumonia in children aged 2-59 months of age in Uttar Pradesh and Bihar, India, in 2016: An indirect estimation.

INTRODUCTION: Community Acquired Pneumonia (CAP) is the leading cause of mortality in children younger than five years of age in developing countries, including India. Hence, this prospective study was performed to estimate the incidence of CAP in children (2-59 months)in four districts of Northern India. METHODS: A cross-sectional survey in rural Lucknow was conducted using cluster sampling technique to assess the proportion of CAP cases that were hospitalized in last 12 months (hospitalization fraction). Another prospective study was done to assess number of hospitalized CAP cases in same districts in 2016. For this, a surveillance network of hospitals that admitted children was established. Cases with WHO-defined CAP with less than 14 days of illness were eligible for inclusion. Informed written parental consent was obtained. A mathematical model was developed to estimate the incidence of CAP in each district, taking into account number of cases hospitalized in one year, assuming it to be equal to hospitalization fraction and using Lucknow district as reference, correcting for child-population per hospital for each district. Population census data of 2011 was taken as denominator. RESULTS: In cross-sectional survey (February to May 2016), 3351 children (2-59 months) from 240 villages were included. Of these 24.58% (824/3351) children suffered from CAP in last 12 months and out of these 4% (33/824) children were hospitalized. Computed incidence of CAP per 1000 child-year for Lucknow was 86.50 (95%CI: 85.72-87.29); Etawah 177.01(95%CI: 175.44-178.58); Patna 207.78 (95%CI: 207.20-208.37) and Darbhanga 221.18 (95%CI: 220.40-221.97). Infants (2-11 months)had almost five to ten times higher incidence of CAP than those in 12-59 months age category. CONCLUSIONS: Incidence of CAP in Uttar Pradesh and Bihar is high, being much higher in infants. Hence there is an urgent need for introduction of preventive strategies, improving health seeking behavior and quality of care for CAP.

To assess the effectiveness of various communication strategies for improving childhood pneumonia case management: study protocol of a community based behavioral open labeled trial in rural Lucknow, Uttar Pradesh, India.

BACKGROUND: Community Acquired Pneumonia (CAP) is the leading cause of childhood morbidity and mortality worldwide including India. Many of these deaths can be averted by creating awareness in community about early symptoms of CAP and by ensuring availability of round the clock, quality health care. The objective was to assess the effectiveness of an innovative package of orienting doctors and community health workers about community perceptions on CAP barriers to qualified health care seeking, plus infrastructural strengthening by (i) providing "Pneumonia Drug Kit" (PDK) (ii) establishing "Pneumonia Management Corner" (PMC) at additional primary health center (PHCs) and (iii) "Pneumonia Management Unit" (PMU) at Community health center (CHCs) along with one of 4 different behavior change communication interventions: 1. Organizing Childhood Pneumonia Awareness Sessions (PAS) for caregivers of children < 5 years of age during a routine immunization day at PHCs and CHCs by Auxillary Nurse Midwives (ANM) 2. Organizing PAS on Village Health and Nutrition Day only once a month in villages by Accredited Social Health Activist (ASHA) 3. Combination of both Interventions 1 & 2 4. Usual Care as measured by number of clinical pneumonia cases-treated by ANM/doctors with PDK or treated at either PMC or PMU. METHODS: Prospective community based open labeled behavioral trial (2 by 2 factorial design) conducted in 8 rural blocks of Lucknow district. Community survey will be done by multistage cluster sampling to collect information on changes in types of health care providers' service utilization for ARI/CAP pre and post intervention. DISCUSSION: CAP is one of the leading killers of childhood deaths worldwide. Studies have reported that recognition of pneumonia and its danger signs is poor among caregivers. The proposed study will assess effectiveness of various communication strategies for improving childhood pneumonia case management interventions at mother/community level, health worker and health center level. The project will generate demand and improve supply of quality of care of CAP and thus result in reduced mortality in Lucknow district. Since the work will be done in partnership with government, it can be scaled up. TRIAL REGISTRATION: This study has been registered retrospectively in the AEARCT Registry and the registration number is: AEARCTR-0003137 .

Dynamic contrast-enhanced (DCE) MRI derived kinetic perfusion indices may help predicting seizure control in single calcified neurocysticercosis.

BACKGROUND: The factors responsible for seizure recurrence in patients with Solitary calcified neurocysticercosis (NCC) are not well understood. Blood brain barrier (BBB) breach may be associated with seizure recurrence. Dynamic contrast enhanced (DCE) MRI derived indices kep, ktrans and ve are useful in quantifying BBB permeability. In this study, we assessed the possible role of DCE-MRI and matrix metalloproteinases (MMP)-9 levels in predicting seizure recurrence. METHODS: In this prospective-observational study, patients with new-onset seizures and a solitary calcified NCC were included. DCE-MRI was done to quantify BBB integrity. DCE-MRI parameters were measured as kep, ktrans and ve. MMP-9 levels were estimated. Patients were followed for 1 year, when DCE-MRI and MMP-9 levels were repeated. Patients were classified into two groups on the basis of seizure recurrence, which was defined as the recurrence of an episode of seizure at least 1 week after the initiation of the anti-epileptic drugs. Logistic regression analysis was done. RESULTS: At 1-year of follow up, 8 out of 32 patients had seizure recurrence. Baseline DCE-MRI derived kep (p = 0.015) and MMP-9 levels (p = 0.019) were significantly higher in the seizure "recurrence" group compared with the "no recurrence" group. On within-group analysis, a significant increase in kep (p = 0.012), ve (p = 0.012), and MMP-9 levels (p = 0.017) was observed in the seizure "recurrence" group while a decrease was seen in ve and MMP-9 levels in the "no recurrence" group. CONCLUSION: Higher values of DCE-MRI indices and MMP-9 levels, with a corresponding trend in the follow-up, can be useful in predicting lesions with a higher propensity for seizure recurrence.

Minimal hepatic encephalopathy in children with chronic liver disease: Prevalence, pathogenesis and magnetic resonance-based diagnosis.

BACKGROUND & AIMS: Data on minimal hepatic encephalopathy (MHE) in children is scarce. We aimed to study MHE in children with chronic liver disease (CLD) and to validate non-invasive objective tests which can assist in its diagnosis. METHODS: We evaluated 67 children with CLD (38 boys; age 13 [7-18] years) and 37 healthy children to determine the prevalence of MHE. We also assessed the correlation of MHE with changes in brain metabolites by magnetic resonance spectroscopy (1HMRS), diffusion tensor imaging (DTI) derived metrics, blood ammonia and inflammatory cytokines (interleukin-6 [IL6], tumor necrosis factor alpha [TNF-α]). In addition, the accuracy of MR-based investigations for diagnosis of MHE in comparison to neuropsychological tests was analysed. RESULTS: Thirty-four (50.7%) children with CLD had MHE on neuropsychological tests. MHE patients had higher BA (30.5 [6-74] vs. 14 [6-66]µmol/L; p=0.02), IL-6 (8.3 [4.7-28.7] vs. 7.6 [4.7-20.7]pg/ml; p=0.4) and TNF-α (17.8 [7.8-65.5] vs. 12.8 [7.5-35]pg/ml; p=0.06) than No-MHE. 1HMRS showed higher glutamine (2.6 [2.1-3.3] vs. 2.4 [2.0-3.1]; p=0.02), and lower choline (0.20 [0.14-0.25] vs. 0.22 [0.17-0.28]; p=0.1) and myo-inositol (0.25 [0.14-0.41] vs. 0.29 [0.21-0.66]; p=0.2) in MHE patients than those without MHE. Mean diffusivity (MD) on DTI was significantly higher in 6/11 brain areas in patients with MHE vs. no MHE. Brain glutamine had a significant positive correlation with blood ammonia, IL-6, TNF-α and MD of various brain regions. Neuropsychological tests showed a negative correlation with blood ammonia, IL6, TNF-α, glutamine and MD. Frontal white matter MD had a sensitivity and specificity of 73.5% and 100% for diagnosing MHE. CONCLUSIONS: In children with CLD, 50% have MHE. There is a significant positive correlation between markers of hyperammonemia, inflammation and brain edema and these correlate negatively with neuropsychological tests. MD on DTI is a reliable tool for diagnosing MHE. LAY SUMMARY: Fifty percent of children with chronic liver disease develop minimal hepatic encephalopathy (MHE) and perform poorly on neuropsychological testing. These children have raised blood ammonia, inflammatory cytokines and mild cerebral edema on diffusion tensor imaging as compared to children without MHE. The higher the ammonia, inflammatory cytokines and cerebral edema levels the poorer the performance on neuropsychological assessment. The estimation of mean diffusivity on diffusion tensor imaging is an objective and reliable method for diagnosing MHE.

Intermediate outcomes of rheumatic mitral stenosis post-balloon mitral valvotomy.

BACKGROUND: Balloon mitral valvotomy is a standard therapeutic modality for managing rheumatic mitral stenosis. Data on intermediate outcomes of this procedure are limited. Thus we investigated the intermediate outcome after balloon mitral valvotomy performed at a large tertiary center in India. METHODS: Case records and follow-up data of 2330 patients who underwent valvotomy from June 1999 to December 2005 were retrieved from the hospital information system and analyzed. RESULTS: The median age of the patients was 32 ± 11 years, 1363 were female including 36 who were pregnant, and 379 were in atrial fibrillation. Follow-up ranged from 1 to 14 years (mean 4.5 years, median 4.0 years). The procedural success rate was 93%. Atrial fibrillation, higher functional class, and worse valve morphology were independent predictors of a poor procedural outcome. Patients with sinus rhythm had better event-free survival (10.43 years, 95% confidence interval: 10.1-10.7) compared to those with atrial fibrillation (8.17 years, 95% confidence interval: 7.5-8.8). Patients who achieved a valve area >1.75 cm(2) had a better event-free survival (11.7 years, 95% confidence interval: 11.4-12.0) than those with a valve area of 1.5-1.74 cm(2) (9.3 years, 95% confidence interval: 9.0-9.7). On multivariate analysis, higher functional class, worse valve morphology, and new significant mitral regurgitation were predictors of a poor outcome. Achieved mitral valve area >1.75 cm(2) was an independent predictor of a good outcome. CONCLUSION: Patients with sinus rhythm, less gross valve deformity, and a post-balloon mitral valvotomy area >1.75 cm(2) had better intermediate outcomes.

Cytokines, MMP-2, and MMP-9 levels in patients with a solitary cysticercus granuloma.

OBJECTIVES: In patients with neurocysticercosis, perilesional inflammatory activity is thought to be responsible for seizures. This study was designed to evaluate the serum and cerebrospinal fluid (CSF) concentrations of cytokines as well as matrix metalloproteinase (MMP)-2 and MMP-9 in patients with a solitary cysticercus granuloma. MATERIALS AND METHODS: The study included 47 patients suffering from seizures in whom a solitary cysticercus granuloma was detected on a computed tomography (CT) scan. The study also included 47 control subjects. Their serum and cerebrospinal fluid (CSF) was analysed for cytokines and MMP levels. A follow-up CT was performed after 6 months. RESULTS: The median levels of cytokines, interleukin (IL)-1ß, IL-5, IL-6, IL-10 and tissue necrosis factor (TNF)-α, MMP-2 and MMP-9 were significantly elevated, both in the serum and CSF of patients having an intracerebral solitary cysticercus granuloma, in comparison to that of controls. The follow-up CT revealed that in 27 patients, the lesions were calcified and in 5 patients, there was complete resolution of the lesions. In 15 patients, the lesions remained unchanged. Higher baseline CSF MMP-2 and TNF-α level were seen in patients with persisting lesions. Higher serum baseline MMP-2, IL-6 and a low CSF IL-10 level were seen in patients with complete resolution of the granuloma. A high baseline IL-1ß level was associated with a calcified lesion. Fourteen patients had recurrence of seizures. A high baseline serum TNF-α level was independently associated with seizure recurrence (P = 0.021, OR = 1.041, CI = 1.006 to 1.078). CONCLUSION: In patients with a solitary cysticercus granuloma, cytokines and matrix metalloproteinases in the CSF and serum are elevated. Different patterns of immunological changes were observed in patients following resolution or calcification of the lesion.

Diffusion tensor tractography and neuropsychological assessment in patients with vitamin B12 deficiency.

INTRODUCTION: Structural imaging of the brain does not demonstrate any changes in a vast majority of patients with vitamin B12 deficiency, even in advanced stages. In this study, we aimed to assess and correlate the functional integrity of the brain fiber tracts using diffusion tensor tractography with neuropsychological examination in patients with vitamin B12 deficiency. METHODS: The study was conducted at two tertiary care centers. Thirty-two patients with vitamin B12 deficiency were enrolled and subjected to diffusion tensor tractography, as an extension of diffusion tensor imaging, and neuropsychological assessment. Tests of significance were done to detect changes, pre- and post-vitamin B12 supplementation in the diffusivity parameters (fractional anisotropy and mean diffusivity) and the neuropsychological test scores. RESULTS: Statistically significant changes were observed in the diffusivity parameters and the neuropsychological test scores between the controls and the patients with vitamin B12deficiency in the pre- and post-treatment phases. CONCLUSIONS: This is the first study to evaluate the diffusion tensor tractography (DTT) parameters in the light of clinical neuropsychological assessment in patients with vitamin B12 deficiency. Utilization of DTT parameters may antedate structural changes and may quantify the neurocognitive deficits.

DTI correlates of cognition in term children with spastic diplegic cerebral palsy.

BACKGROUND AND AIMS: Presently, there is no published study that shows association between cognition and white matter injury in spastic cerebral palsy. We aimed to correlate cognitive functions with diffusion tensor imaging derived metrics in spastic diplegic children. METHODS: Twenty-two term children with spastic diplegia and 22 healthy controls were recruited. All patients were graded on the basis of gross motor function. The Indian children intelligence Test was used to quantify cognition and diffusion tensor imaging was used to quantify microstructural changes in various white matter regions. Diffusion tensor imaging metrics were quantified by placing regions of interests in different white matter regions like corona radiata, anterior limb of internal capsule, posterior limb of internal capsule, mid brain, pons, medulla, genu, splenium, temporal white matter, parietal white matter, frontal white matter and occipital white matter. RESULTS: Spastic diplegic children showed significantly lower neuropsychological test scores as compared to controls. A significantly decreased fractional anisotropy values were observed in corona radiata, anterior limb of internal capsule, posterior limb of internal capsule, mid brain, pons, medulla, genu, splenium and occipital white matter; however significantly increased mean diffusivity values were observed in corona radiata, anterior limb of internal capsule, posterior limb of internal capsule, mid brain, pons and genu in spastic diplegic as compared to controls. A significant positive correlation in fractional anisotropy and negative correlation in mean diffusivity was observed with neuropsychological test scores. CONCLUSION: These results suggest that these imaging metrics may be used as a biomarker of cognitive functions in term children with spastic diplegia.

Novel corrective equations for complete estimation of human tissue lipids after their partial destruction by perchloric acid pre-treatment: high-resolution (1)H-NMR-based study.

Owing to the small quantity of tissue available in human biopsy specimens, aqueous and lipid components often have to be determined in the same tissue sample. Perchloric acid (PCA) used for the extraction of aqueous metabolites has a deleterious effect on lipid components; the severity of the damage is not known. In this study, human muscle tissue was first treated with PCA to extract aqueous metabolites, and the residue was then used for lipid extraction by conventional methods, i.e. the methods of Folch and Bligh & Dyer and a standardised one using methanol/chloroform (1:3, v/v) used in our laboratory. A (1)H-NMR spectrum was obtained for each lipid extract. Lipid was quantified by measuring the integral area of N(+)-(CH(3))(3) signals of phospholipids (PLs). Triacylglycerol (TG) and cholesterol (CHOL) were quantified using the -CH(2)- signals of glycerol and the C18 methyl signal, respectively. This study shows that prior use of PCA caused marked attenuation of TG, PL, and CHOL. This was confirmed by recovery experiments and observation of the direct effect of PCA on the standard lipid components. On the basis of the quantity of lipid lost in each case, three novel equations (with respect to TG, PL, and CHOL) were derived. Application of these equations to lipid quantities estimated in different pathological tissues after PCA pre-treatment produced values equivalent to those estimated without PCA use. This study conclusively shows that PCA pre-treatment damages all three lipid moieties, TG, PL, and CHOL. When PCA is used in a fixed ratio to the tissue, the lipid damage is also proportional and correctable by statistically derived equations. These equations will be useful in human biopsy specimens where aqueous and lipid components have to be studied using the same tissue sample because of the small quantity available.

Prednisolone in Duchenne muscular dystrophy with imminent loss of ambulation.

An open controlled trial of 0.75 mg/Kg/day prednisolone was conducted at a stage when the patients had started falling several times in a day and stopped on their attaining a chair bound stage, thus minimising the total period of steroid therapy. Out of the 67 DMD patients enrolled in this study, 44 were put on prednisolone therapy and 23 served as controls. All patients were followed-up at two-monthly intervals for two years and thereafter they continued to take their respective medications till their chair-bound stage; then the drug was gradually withdrawn. In the treatment group 24 patients could not continue the trial because of adverse effects - 14 due to excessive obesity, 3 due to measles, 4 due to pulmonary tuberculosis, 2 due to recurrent throat and chest infection and 1 due to an unexplained high leukocyte count. Of the remaining 20 patients in the treatment group, steroid therapy was stopped in 5 patients as there was no improvement in power in six months. Fifteen patients in the treatment group and 19 patients in the control group could be followed regularly for 2 years and then up to chair-bound stage. Outcome parameters included fall frequency, peak expiratory flow rate, limb muscle power, ability to lift weights, time taken in getting up from squatting position, walking 9 metres and climbing 13 stairs. Maximum improvement was noted between 2 and 4 months while mild improvement in some parameters continued up to six months. All parameters remained stabilised for 1 year or so, after which there was slight deterioration. Deterioration at 2 years was, however, less than the natural course of events noted in control patients. Prednisolone treated patients and controls became chair bound at the mean age of 169 +/- 9 and 132 +/- 8 months respectively. Till the ideal stage of the disease and the type or dosage of starting steroid therapy is defined by specially designed studies, 0.75 mg/Kg/day prednisolone therapy may be started in DMD patients at the stage of frequent falls ( > 10 / day) on walking or increased get-up time ( > 10 s) as observed while testing Gowers' sign; this improves muscle power and timing of motor performance within 2-4 months of onset of therapy in about 75% of those who tolerate this therapy, with a possible gain of approximately 3 years in terms of independent walking.