An integrated approach to the structural characterization, long-term toxicological and anti-inflammatory evaluation of Pterospermum rubiginosum bark extract.

ETHNOPHARMACOLOGICAL RELEVANCE OF STUDY: Pterospermum rubiginosum is an evergreen plant in Western Ghats, India, used by traditional tribal healers due to its excellent biological potential for treating inflammation and pain relief procedures. The bark extract is also consumed to relieve the inflammatory changes at the bone fractured site. The traditional medicinal plant in India have to be characterized for its diverse phytochemical moieties, its interactive multiple target sites, and to reveal the hidden molecular mechanism behind the biological potency. AIM OF THE STUDY: The study focussed on plant material characterization, computational analysis (prediction study), toxicological screening (In vivo), and anti-inflammatory evaluation of P. rubiginosum methanolic bark extracts (PRME) in LPS-induced RAW 264.7 cells. MATERIALS AND METHODS: The pure compound isolation of PRME and their biological interactions were used to predict the bioactive components, molecular targets, and molecular pathways of PRME in inhibiting inflammatory mediators. The anti-inflammatory effects of PRME extract were evaluated in the lipopolysaccharide (LPS)-induced RAW264.7 macrophage cell model. The toxicity evaluation of PRME was performed in healthy 30 Sprague-Dawley experimental rats, were randomly divided into five groups for toxicological evaluation for 90 days. The tissue levels of oxidative stress and organ toxicity markers were measured using the ELISA method. Nuclear magnetic resonance spectroscopy (NMR) was performed to characterize the bioactive molecules. RESULTS: Structural characterization revealed the presence of vanillic acid, 4-O-methyl gallic acid, E-resveratrol, gallocatechin, 4'-O-methyl gallocatechin, and catechin. Molecular docking of NF-kB exhibited significant interactions with vanillic acid and 4-O-methyl gallic acid with binding energy -351.159 Kcal/Mol and -326.5505 Kcal/Mol, respectively. The PRME-treated animals showed an increase in total GPx and antioxidant levels (SOD and catalase). Histopathological examination revealed no variation in the liver, renal and splenic tissue's cellular pattern. PRME inhibited the pro-inflammatory parameters (IL-1ß, IL-6, and TNF-α) in LPS-induced RAW 264.7 cells. The protein level of TNF-α and NF-kB protein expression study brought out a notable reduction and exhibited a good correlation with the gene expression study. CONCLUSION: The current study establishes the therapeutic potential of PRME as an effective inhibitory agent against LPS-activated RAW 264.7 cells induced inflammatory mediators. Long-term toxicity evaluation on SD rats confirmed the non-toxic nature of PRME up to 250mg/body weight for 3 months.

Characterization and distribution of microplastics in estuarine surface sediments, Kayamkulam estuary, southwest coast of India.

The present study aims to formulate the characterization and distribution of microplastic in the estuarine surface sediments of Kayamkulam estuary, southwest coast of India. The sediments were dominated by fibre and film shaped microplastic substances. The surface sediments were dominated by <1000 µm microplastics. The composition of microplastics in descending order was as follows: polyester > polypropylene > polyethylene. The distribution of microplastics was significantly higher than that observed in the other study regions, except for Pearl river estuary and Guanabara Bay. The distribution of microplastics was chiefly controlled by estuarine inundating water and the distance of the sampling site from the open sea.

Anti-inflammatory effect and mechanism of action of ellagic acid-3,3',4-trimethoxy-4'-O-α-L-rhamnopyranoside isolated from Hopea parviflora in lipopolysaccharide-stimulated RAW 264.7 macrophages.

Phytochemical investigation of the stem bark of Hopea parviflora resulted in the isolation of 9 compounds; which includes friedelin (1), friedelin-3ß-ol (2), (-)-ampelopsin A (3), (-)-ɛ-viniferin (4), (-)-hopeaphenol (5), vaticaphenol A (6), 2,4,8-trihydroxyphenanthrene-2-O-glucoside (7), ellagic acid-3,3',4-trimethoxy-4'-O-α-L-rhamnopyranoside (8) and ß-sitosterol-ß-D-glucoside (9). Among them, compounds 1, 2, 6, 7, 8 and 9 are isolated for the first time from this species. Further, we evaluated the anti-inflammatory activity of compounds 4, 5, 6, 7 and 8. In this study, compound 8 inhibited the activity of proinflammatory mediators like NO, TNF-α, IL-6, 5-LOX and COX-2, also promoted the action of anti-inflammatory mediator like IL-10 via inhibition of the NF-κB pathway in LPS-stimulated RAW 264.7 macrophages.

Correlates of High-Risk and Low-Risk Alcohol Use among College Students in Kerala, India.

This study describes the prevalence and correlates of alcohol use among college students in Ernakulam, Kerala State, India. A total of 5784 students from 58 colleges selected by stratified random sampling completed a questionnaire incorporating standardized instruments. The prevalence of lifetime alcohol use was 21.4% with a male predominance. Among users, low-risk, hazardous, and dependent use were 80.2%, 18.3% and 0.9% respectively. Initiation was mostly with friends (45.3%). Both low-risk and high-risk alcohol users (hazardous and dependent users), in comparison to abstainers, had higher odds of being older, non-Muslim, having a part-time job, using other substances, and exposure to sexual abuse. Students who reported low-risk use also had an urban background, more severe psychological distress and suicidal thoughts, while high-risk users had attention deficit hyperactivity disorder (ADHD) symptoms. Students who reported high-risk use compared to low-risk users had higher odds of having a part-time job, tobacco use, and ADHD symptoms. Alcohol use among college students is common, with both low- and high-risk drinking associated with significant morbidity. This study highlights the need to promote public health policies to target and prevent all patterns of alcohol use among young people.

Effects of a new synthetic zerumbone pendant derivative (ZPD) on apoptosis induction and anti-migratory effects in human cervical cancer cells.

A newly synthesised zerumbone pendant derivative (ZPD) was studied in human cervical cancer cells (HeLa) for its anticancer properties. ZPD significantly inhibited the growth of human cervical cancer cells with a GI50 value of 6.35 ± 1.30 µM, which also induced morphological changes and apoptosis in a dose-dependent manner. Our data indicated that ZPD actively encouraged programmed cell death in HeLa cells which were confirmed by DNA fragmentation, phosphatidylserine translocation, increased activity of caspase 3, upregulation of the expression of the pro-apoptotic protein Bax, cleaved PARP, cleaved caspase 3 and downregulation of anti-apoptotic protein Bcl-2. ZPD also inhibited cell migration of HeLa cells, decreasing the production of MMP-2,-9 and downregulation of expression of MMPs and pro-angiogenic factor VEGF. Also it is nontoxic to normal rat cardiac myoblasts. Overall, ZPD is a promising candidate for inducing cytotoxicity, apoptosis and anti-migratory effects in cervical cancer cells.

Novel glycoconjugated squaraine dyes for selective optical imaging of cancer cells.

Novel glycoconjugated squaraine dyes were synthesized, which exhibited excellent internalization in tumor cells and are potent fluorogenic imaging probes for tumor selective optical imaging.

Prevalence and correlates of obsessive-compulsive disorder and subthreshold obsessive-compulsive disorder among college students in Kerala, India.

CONTEXT: There are scarce data on the prevalence of adult obsessive-compulsive disorder (OCD) in India. AIMS: The aim was to study the point prevalence of OCD and subthreshold OCD and its psychosocial correlates among college students in the district of Ernakulam, Kerala, India. SETTINGS AND DESIGN: A cross-sectional survey of 5784 students of the age range of 18-25 years from 58 colleges was conducted. MATERIALS AND METHODS: Students were self-administered the OCD subsection of the Clinical Interview Schedule-Revised, the Composite International Diagnostic Interview for obsessive-compulsive symptoms (OCSs), and other relevant instruments to identify OCD, subthreshold OCD, and related clinical measures. STATISTICAL ANALYSIS: The point prevalence of OCD and subthreshold OCD was determined. Categorical variables were compared using Chi-square/Fisher's exact tests as necessary. Differences between means were compared using the ANOVA. RESULTS: The point prevalence of OCD was 3.3% (males = 3.5%; females = 3.2%). 8.5% students (males = 9.9%; females = 7.7%) fulfilled criteria of subthreshold OCD. Taboo thoughts (67.1%) and mental rituals (57.4%) were the most common symptoms in OCD subjects. Compared to those without obsessive-compulsive symptoms (OCSs), those with OCD and subthreshold OCD were more likely to have lifetime tobacco and alcohol use, psychological distress, suicidality, sexual abuse, and higher attention-deficit/hyperactivity disorder symptom scores. Subjects with subthreshold OCD were comparable to those with OCD except that OCD subjects had higher psychological distress scores and academic failures. CONCLUSIONS: OCD and subthreshold OCD are not uncommon in the community, both being associated with significant comorbidity. Hence, it is imperative that both are identified and treated in the community because of associated morbidity.

Study of stability and biophysical characterization of ranibizumab and aflibercept.

The anti-vascular endothelial growth factor (VEGF) agents such as ranibizumab (Lucentis®) and aflibercept (EyLea®) are currently used as monthly or bimonthly intravitreal injections to treat potentially retinal diseases such as wet age-related macular degeneration (AMD) or diabetic macular edema (DME). Because of the complications associated with repeated intra-vitreal injections, there is considerable interest in developing a sustained delivery system. The purpose of this study was to examine the stability of both therapeutic proteins under physiological conditions as well as when incorporated into drug delivery systems (DDS). First, thermotropic properties in physiological conditions and at different pH values were evaluated by differential scanning calorimetry (DSC) to determine the protein denaturation temperature. Second, the effects of pH and incubation time on conformational changes and aggregation were evaluated by circular dichroism (CD), steady-state tryptophan fluorescence spectroscopy, and size-exclusion chromatography (SEC). Also, the ability of both proteins to bind to VEGF was tested in the aforementioned experimental conditions for up to 30 days. Finally, we investigated the stability of both proteins after a rapid screening method that simulates the first homogenizing step during the protein microencapsulation process. This method allowed the development of stable ranibizumab and aflibercept formulations that may be useful to entrap these proteins into microparticles selecting the most convenient organic solvent and protein stabilizers.

Reduced toxicity, myeloablative conditioning with BU, fludarabine, alemtuzumab and SCT from sibling donors in children with sickle cell disease.

BU and CY (BU/CY; 200 mg/kg) before HLA-matched sibling allo-SCT in children with sickle cell disease (SCD) is associated with ~85% EFS but is limited by the acute and late effects of BU/CY myeloablative conditioning. Alternatives include reduced toxicity but more immunosuppressive conditioning. We investigated in a prospective single institutional study, the safety and efficacy of a reduced-toxicity conditioning (RTC) regimen of BU 12.8-16 mg/kg, fludarabine 180 mg/m(2), alemtuzumab 54 mg/m(2) (BFA) before HLA-matched sibling donor transplantation in pediatric recipients with symptomatic SCD. Eighteen patients, median age 8.9 years (2.3-20.2), M/F 15/3, 15 sibling BM and 3 sibling cord blood (CB) were transplanted. Mean whole blood and erythroid donor chimerism was 91% and 88%, at days +100 and +365, respectively. Probability of grade II-IV acute GVHD was 17%. Two-year EFS and OS were both 100%. Neurological, pulmonary and cardiovascular function were stable or improved at 2 years. BFA RTC and HLA-matched sibling BM and CB allo-SCT in pediatric recipients result in excellent EFS, long-term donor chimerism, low incidence of GVHD and stable/improved organ function.

National epilepsy surgery support activity.

While there are over one million people with drug-resistant epilepsy in India, today, there are only a handful of centers equipped to undertake presurgical evaluation and epilepsy surgery. The only solution to overcome this large surgical treatment gap is to establish comprehensive epilepsy care centers across the country that are capable of evaluating and selecting the patients for epilepsy surgery with the locally available technology and in a cost-effective manner. The National Epilepsy Surgery Support Activity (NESSA) aims to provide proper guidance and support in establishing epilepsy surgery programs across India and in neighboring resource-poor countries, and in sustaining them.

Regional brain blood flow in mouse: quantitative measurement using a single-pass radio-tracer method and a mathematical algorithm.

We have developed a reliable experimental method for measuring local regional cerebral blood flows in anesthetized mice. This method is an extension of the well-established single-pass dual-label indicator method for simultaneously measuring blood flow and glucose influx in rat brains. C57BL6J mice (n = 10) were anesthetized and regional blood flows (ml/min/g) were measured using the radio-tracer method. To test the sensitivity of this method we used a mathematical algorithm to predict the blood flows and compared the two sets of results.Measured regional blood flows between 0.7 and 1.7 ml/min/g were similar to those we have previously reported in the rat. The predicted blood flows using an assumed linearly increasing arterial tracer concentration-versus-time profile (that is, a ramp) were similar to the values measured in the physiological experiments (R(2) 0.99; slope 0.91). Thus,measurements of local regional cerebral blood flow in anesthetized mice using a single-pass radio-tracer method appear to be reliable.

Rasmussen's encephalitis: experience from a developing country based on a group of medically and surgically treated patients.

PURPOSE: To describe the attributes of patients with Rasmussen's encephalitis (RE) seen in a tertiary epilepsy referral center in southern India and to enquire factors helpful in predicting responsiveness to immunotherapy. METHODS: We diagnosed RE based on the European consensus criteria. To assess the factors that could potentially predict the natural course and therapeutic outcome, we subcategorized our patients according to age at onset ( 6 years), duration from onset to presentation ( 2 years), immunotherapy versus surgery, and early (

Candida spp. other than Candida albicans: a major cause of fungaemia in a tertiary care centre.

This study was conducted to determine the frequency of different Candida spp. isolated from different parts of the hospital, associated risk factors and mortality rate. A total of 59 cases were selected for prospective analysis over a period of one and half years. Blood samples collected were processed by BACTEC (9240) method. Candidaemia was diagnosed by positive blood culture at least from two blood culture samples or from a clinically significant single blood culture sample. Candida spp. were identified by standard techniques. Most frequent isolates were C. tropicalis (35.6%), C. parapsilosis (28.8%), C. glabrata (11.9%) and C. pelliculosa (11.9%). Candida albicans was isolated only in 3.4% cases. Neonatology department accounted for highest number of isolates (27.1%), followed by gastrointestinal surgery (15.3%) and cardiac surgery (13.6%). Mortality was noted in 16.9%. Probable risk factors determined were intensive care unit stay (74.6%), antibiotic therapy (50.8%), central line (42.4%), urinary catheter (32.2%), ventilator (23.7%), malignancy (20.3%) and abdominal surgery (15.3%).

Risk factors for acute ischaemic stroke in young adults in South India.

BACKGROUND: Stroke is a leading cause of death and disability in developing countries, afflicting individuals at a young age. The contribution of established vascular risk factors to ischaemic stroke in young adults has not been evaluated systematically in Indians. METHODS: We conducted a case control study in 214 South Indian patients with first acute ischaemic stroke that occurred between the ages of 15 and 45 years, 99 age and sex matched hospital controls and 96 community controls. We compared the prevalence of the following risk factors: smoking, elevated blood pressure, high fasting blood glucose and abnormal lipids. RESULTS: Compared with community controls, stroke patients had a higher prevalence of smoking (multivariable adjusted odds ratio (OR) 7.77, 95% CI 1.93 to 31.27), higher systolic blood pressure (OR per SD increment of 1.88, 95% CI 1.01 to 3.49) and fasting blood glucose (OR per SD increment of 4.55, 95% CI 1.63 to 12.67), but lower high density lipoprotein (HDL) cholesterol (OR per SD increment of 0.17, 95% CI 0.09 to 0.30). Compared with hospital controls, stroke patients had a higher prevalence of smoking (OR 3.95, 95% CI 1.61 to 9.71) and lower HDL cholesterol (OR per SD increment 0.27, 95% CI 0.17 to 0.44). The presence of > or = 3 metabolic syndrome components was associated strongly with stroke (OR 4.76, 95% CI 1.93 to 11.76; OR 2.09, 95% CI 1.06 to 4.13) compared with community and hospital controls. CONCLUSIONS: Key components of the metabolic syndrome and smoking are associated with ischaemic stroke in young South Indian adults. Our observations underscore the importance of targeting adolescents and young adults for screening and prevention to reduce the burden of ischaemic stroke in young adults.

Hemimegalencephalic appearance of normal hemisphere in unilateral heterotopia and absent corpus callosum.

We report two patients with medically refractory epilepsy who had MRI evidence of unilateral subcortical nodular heterotopia and agenesis of corpus callosum. The abnormal hemisphere was small, whereas the contralateral normal hemisphere appeared large and crossed the midline. Although the normal hemisphere was initially mistaken for hemimegalencephaly, there were no typical radiological features. Moreover, the electroencephalographic abnormalities lateralized to the hemisphere showing heterotopia. Because contralateral hemispheric abnormalities like heterotopia, hemimicrencephaly, and hemimegalencephaly can occur in patients with hemispheric heterotopias, we emphasize the importance of careful scrutiny of the contralateral hemisphere in patients with unilateral heterotopia. Absence of typical radiological features and appropriately lateralized electroencephalographic abnormalities will help differentiate the two. This is crucial when planning epilepsy surgery.

Seizure outcome after anterior temporal lobectomy and its predictors in patients with apparent temporal lobe epilepsy and normal MRI.

PURPOSE: Very little reliable information is available regarding the role of anterior temporal lobectomy (ATL), optimal presurgical evaluation strategy, post-ATL seizure outcome, and the factors that predict the outcome in patients with medically refractory temporal lobe epilepsy (TLE) and normal high-resolution magnetic resonance imaging (MRI). To be cost-effective, epilepsy surgery centers in developing countries will have to select candidates for epilepsy surgery by using the locally available technology and expertise. METHODS: We reviewed the electroclinical and pathological characteristics and seizure outcome of 17 patients who underwent ATL for medically refractory TLE after being selected for ATL based on a noninvasive selection protocol without the aid of positron emission tomography (PET) or single-photon emission computed tomography (SPECT), despite a normal preoperative high-resolution MRI. RESULTS: Seven (41%) patients achieved an excellent seizure outcome; five of them were totally seizure free. An additional five (29%) patients had >75% reduction in seizure frequency. The following pre-ATL factors predicted an excellent outcome: antecedent history of febrile seizures, strictly unilateral anterior temporal interictal epileptiform discharges (IEDs), and concordant type 1 ictal EEG pattern. All the five patients with pathologically verified hippocampal formation neuronal loss were seizure free. The presence of posterior temporal, bilateral temporal, and generalized IEDs portended unfavorable post-ATL seizure outcome. CONCLUSIONS: A subgroup of patients destined to have an excellent post-ATL outcome can be selected from MRI-negative TLE patients by using history and scalp-recorded interictal and ictal EEG data. The attributes of these patients are antecedent history of febrile seizures, strictly unilateral anterior IEDs, and concordant type 1 ictal EEG pattern.

Antiulcer activity of Utleria salicifolia rhizome extract.

The effect of 50% ethanolic extract of Utleria salicifolia (USE) was assessed in different acute and chronic gastric ulcer models in rats. USE, 50-200 mg/kg administered orally, twice daily for 5 days showed dose-dependent ulcer protective effect in pylorus ligation (14.48-51.03% protection, P < 0.5 to P < 0.01), aspirin (28.80-56.52% protection, P < 0.5 to P < 0.001), ethanol (13.22-60.74% protection, P < 0.5 to P < 0.001), cold-restraint stress (21.22-77.14% protection, P < 0.05 to P < 0.001), and acetic acid (20.0-84.37% protection, P < 0.5 to P < 0.001)-induced acute and chronic ulcers. USE also significantly (P < 0.001) reduced the ulcer incidence (50 and 10%) and severity (67.83 and 91.34% protection) of duodenal ulcer, induced by cysteamine. Besides USE offered protection (53.52 and 60.58%) against ethanol-induced depletion of gastric wall mucus. However, USE reduced the ulcer index with significant decrease in plasma corticosterone (25.53 and 39.52% protection, P < 0.1 and P < 0.05), lipid peroxidation (18.75 and 47.92% protection, P < 0.01 and P < 0.001), superoxide dismutase (15.80 and 26.61% protection, P < 0.05 and P < 0.001) and increased in catalase (28.42 and 71.0% protection, P < 0.05 and P < 0.001) activity, respectively. Preliminary phytochemical screening of the USE gave the positive test for steroids, alkaloids, terpenoids, saponins and tannins. The HPTLC studies in the toluene: ethyl acetate: formic acid and the densitometric scanning at 254 nm gave three major spots with area corresponding to 28.16, 17.17, and 13.79% at 0.69, 0.78, and 0.88 R(f) values, respectively. The results indicate that USE possesses antiulcer activity.

Differential diagnosis of patients with intracranial sinus venous thrombosis related isolated intracranial hypertension from those with idiopathic intracranial hypertension.

In patients presenting with intracranial hypertension without hydrocephalus, mass lesions, and with normal cerebrospinal fluid (CSF) composition (pseudotumor cerebri syndrome), the diagnosis of intracranial sinus venous thrombosis (ISVT) has crucial etiological, therapeutic and prognostic implications. Utilizing two well-defined groups of pseudotumor cerebri patients, one with magnetic resonance imaging (MRI) or angiography confirmed ISVT (17 patients) and the other in whom ISVT has been excluded (idiopathic intracranial hypertension [IIH], 27 patients), we investigated the characteristics that might be helpful in distinguishing them. No clinical or auxiliary findings differed between the ISVT and IIH groups except for female gender and lower CSF protein level, which were significantly associated with the latter. While the syndrome pseudotumor cerebri could be due to multiple causes including ISVT, the term IIH should be restricted for patients with isolated intracranial hypertension attributable to no other neurological or systemic disease. Since CT frequently misses ISVT, patients with pseudotumor cerebri syndrome should undergo MRI and MR venography before being labeled as IIH. We conclude that Modified Dandy's Diagnostic Criteria of pseudotumor cerebri, formulated prior to MRI era, can no longer be applied for the diagnosis of IIH.

Selection of ideal candidates for epilepsy surgery in developing countries.

In order to become cost-effective, epilepsy surgery centers in developing countries will have to achieve excellent results by selecting candidates destined to have a seizure-free outcome using locally available limited technology and expertise, without compromising on patient safety. Our experience illustrates that this goal can be accomplished by selecting patients, whose epileptogenic zone can be unquestionably established, based on history, magnetic resonance imaging, and interictal and ictal scalp electroencephalogram findings. Patients with mesial temporal lobe epilepsy, and those with circumscribed potentially epileptogenic lesions belong to this category. A stepwise approach by reserving more difficult to treat patients to later date as experience develops, or by referring them to a better-equipped center, will help each center to understand its capabilities and limitations and to move forward. It would be essential to work with and educate the local public and professionals, if the epilepsy surgery program in a developing region were to have a lasting impact, It is encouraging to note that, despite major challenges, in the last one-decade, several epilepsy centers in the developing world have not only successfully implemented epilepsy surgery programs, but also have produced results comparable to that from developed countries at a fractional cost.

Idiopathic hypertrophic cranial pachymeningitis.

Idiopathic hypertrophic cranial pachymeningitis is a rare form of fibrosing chronic inflammatory process of unknown etiology, which causes thickening of the intracranial dura mater. We present four patients with hypertrophic cranial pachymeningitis who presented with chronic headache and cranial nerve palsies. The diagnosis of idiopathic hypertrophic cranial pachymeningitis was based on neuroimaging findings of thickened enhancing dura, exclusion of known causes and histopathologic findings compatible with nonspecific inflammation in the meningeal biopsies. Corticosteroid therapy was effective in all cases in inducing a complete or partial remission of the neurologic symptoms and signs. We describe the clinical, radiological and pathological features of idiopathic hypertrophic cranial pachymeningitis and discuss the relationship of this entity with other inflammatory fibrosclerotic disorders to explain the pathogenesis. A high index of suspicion, prompt confirmation of the diagnosis by meningeal biopsy, and early institution and long-term maintenance of steroid therapy may help to prevent irreversible neurologic sequelae, especially blindness.