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2012| March | Volume 135 | Issue 3
Online since
May 3, 2012
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REVIEW ARTICLES
Inflammation & apoptosis in spinal cord injury
Ning Zhang, Ying Yin, Sheng-Jie Xu, Yong-Ping Wu, Wei-Shan Chen
March 2012, 135(3):287-296
Spinal cord injury (SCI) consists of a two-steps process involving a primary mechanical injury followed by an inflammatory process and apoptosis. Secondary insult is characterized by further destruction of neuronal and glial cells, and leads to expansion of the damage, so that the paralysis can extend to higher segments. With the identification of mechanisms that either promote or prevent neuronal inflammation and apoptosis come new approaches for preventing and treating neurodegenerative disorders. From a clinical perspective, this article discusses novel targets for the development of therapeutic agents that have the potential to protect the spinal cord from irreversible damage and promote functional recovery.
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SYSTEMATIC REVIEW
Efficacy & safety of iodopovidone pleurodesis: a systematic review & meta-analysis
Ritesh Agarwal, Ajmal Khan, Ashutosh N Aggarwal, Dheeraj Gupta
March 2012, 135(3):297-304
Background & objectives:
Chemical pleurodesis is an accepted therapy for patients with recurrent pleural effusions and pneumothorax. Iodopovidone has been shown to be safe and effective for chemical pleurodesis in several studies. The aim of this systematic review was to update a previously reported meta-analysis on the efficacy and safety of iodopovidone pleurodesis.
Methods:
Two databases MEDLINE and EMBASE were searched for a period (1952-2010), and studies that have reported success rates with iodopovidone pleurodesis were selected. The proportions with 95 per cent confidence interval (CI) were calculated to assess the outcomes in the individual studies and the results were pooled using a random effects model.
Results:
Thirteen eligible studies with 499 patients were included in the mata-analysis. The success rates varied from 70 to 100 per cent in different studies with the pooled success rate being 88.7 per cent (95% CI, 84.1 to 92.1). The success rate was not affected by the method (tube thoracostomy vs. thoracoscopy, 89.6 vs. 94.2%) or the indication of pleurodesis (pleural effusion vs. pneumothorax, 89.2 vs. 94.9%). The only significant complication reported was chest pain of varying degree. Systemic hypotension was reported in six patients across the studies. There were no deaths associated with iodopovidone pleurodesis. Statistical heterogeneity and publication bias were found.
Interpretation & conclusions:
Iodopovidone may be considered a safe and effective agent for chemical pleurodesis in patients with pleural effusions and recurrent pneumothoraces.
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ORIGINAL ARTICLES
Isolation, identification & characterization of
Proteus penneri
- a missed rare pathogen
Janak Kishore
March 2012, 135(3):341-345
Background & objectives:
Indole negative
Proteus
species are invariably incorrectly identified as
P. mirabilis,
missing isolates of
Proteus penneri
.
P. penneri
is an invasive pathogen capable of causing major infectious diseases still seldom reported in individual cases. We report here the isolation, differentiation, characterization and typing of
P. penneri
from patients with different clinical infections.
Methods:
Urine, pus and body fluids collected from patients in intensive care units, wards and out patients departments of a tertiary health care institute from north India were cultured. A total of 61 indole negative
Proteus
isolates were subjected to extended biochemical tests to differentiate and identify
P. penneri
from
P. mirabilis
including failure to produce ornithine decarboxylase (by 0% strains of
P. penneri
and 100% strains of
P. mirabilis
) besides
P. penneri
being uniformly salicin negative, non-utilizer of citrate but ferments sucrose and maltose. Antibiograms and Dienes phenomenon were performed to characterize and type
P. penneri
isolates besides screening for β-lactamase production.
Results:
Eight isolates of
P. penneri
were identified; four from urine, three from abdominal drain-fluid and one from diabetic foot ulcer.
P. penneri
was isolated as the sole pathogen in all patients having underlying disease; post-operatively. Swarming was not seen in the first strain on primary isolation and was poor in strain-4. All eight isolates were biochemically homologous but multi-drug resistant (MDR) with resistance to 6-8 drugs (up to 12). β-lactamase production was seen in three of five isolates while Dienes phenomenon found four distinct types and discriminated strains differing in resistance even with a single drug.
Interpretation & Conclusions:
A few additional biochemical tests identified
P. penneri
isolates; it infected patients with underlying disease and strains were MDR and heterogenous.
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Morphological abnormalities in baseline ECGs in healthy normal volunteers participating in phase I studies
Pooja Hingorani, Mili Natekar, Sheetal Deshmukh, Dilip R Karnad, Snehal Kothari, Dhiraj Narula, Yash Lokhandwala
March 2012, 135(3):322-330
Background & objectives:
Morphological abnormalities in 12-lead electrocardiograms (ECGs) are seen in subgroups of healthy individuals like athletes and air-force personnel. As these populations may not truly represent healthy individuals, we assessed morphological abnormalities in ECG in healthy volunteers participating in phase I studies, who are screened to exclude associated conditions.
Methods:
ECGs from 62 phase I studies analyzed in a central ECG laboratory were pooled. A single drug-free baseline ECG from each subject was reviewed by experienced cardiologists. ECG intervals were measured on five consecutive beats and morphological abnormalities identified using standard guidelines.
Results:
Morphological abnormalities were detected in 25.5 per cent of 3978 healthy volunteers (2495 males, 1483 females; aged 18-76 yr); the presence was higher in males (29.3% vs. 19.2% in females;
P
<0.001). Rhythm abnormalities were the commonest (11.5%) followed by conduction abnormalities (5.9%), axis deviation (4%), ST-T wave changes (3.1%) and chamber enlargement (1.4%). Incomplete right bundle branch block (RBBB), short PR interval and right ventricular hypertrophy were common in young subjects (<20 yr) while atrial fibrillation, first degree atrioventricular block, complete RBBB and left anterior fascicular block were more prevalent in elderly subjects (>65 yr). Prolonged PR interval, RBBB and intraventricular conduction defects were more common in males while sinus tachycardia, short PR interval and non-specific T wave changes were more frequent in females.
Interpretation & Conclusions:
Morphological abnormalities in ECG are commonly seen in healthy volunteers participating in phase I studies; and vary with age and gender. Further studies are required to determine whether these abnormalities persist or if some of these disappear on follow up.
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Antibiotic resistance genes & susceptibility patterns in staphylococci
Nizami Duran, Burcin Ozer, Gulay Gulbol Duran, Yusuf Onlen, Cemil Demir
March 2012, 135(3):389-396
Background & objectives:
This study was carried out to evaluate the association between the antibiotic susceptibility patterns and the antibiotic resistance genes in staphylococcal isolates obtained from various clinical samples of patients attending a teaching hospital in Hatay, Turkey.
Methods:
A total of 298 staphylococci clinical isolates were subjected to antimicrobial susceptibility testing. The genes implicated in resistance to oxacillin (
mecA
), gentamicin (
aac(6')/aph(2''), aph(3')-IIIa, ant(4')-Ia
), erythromycin (
ermA
,
ermB
,
ermC
, and
msrA
), tetracyclin (
tetK, tetM
)
,
and penicillin (
blaZ
) were amplified using multiplex PCR method.
Results:
Methicillin resistance rate among 139
Staphlococcus aureus
isolates was 16.5 and 25.9 per cent of
S. aureus
carried
mecA
gene. Of the 159 CoNS isolates, methicillin resistance rate was 18.9 and 29.6 per cent carried
mecA
gene. Ninety four isolates identified as gentamicin resistant phenotypically
, contained at least one of the gentamicin resistance genes [
aac(6')/aph(2''),
aph(3')-IIIa
,
ant(4')-Ia
], 17 gentamicin-susceptible isolates were found as positive in terms of one or more resistance genes [
aac(6')/aph(2''), aph(3')-IIIa, ant(4')-Ia
] by multiplex PCR. A total of 165 isolates were resistant to erythromycin, and contained at least one of the erythromycin resistance genes (
ermA
,
ermB
,
ermC
and
msrA
). Phenotypically, 106 staphylococcal isolates were resistant to tetracycline, 121 isolates carried either
tetK
or tetM or both resistance genes. The majority of staphylococci tested possessed the
blaZ
gene (89.9%).
Interpretation & Conclusions:
The present results showed that the phenotypic
antibiotic susceptibility patterns
were not similar to those obtained by genotyping done by multiplex PCR. Rapid and reliable methods for antibiotic susceptibility are important to determine the appropriate therapy decisions. Multiplex PCR can be used for confirmation of the results obtained by conventional phenotypic methods, when needed.
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Evaluation of HER-2/
neu
status in breast cancer specimens using immunohistochemistry (IHC) & fluorescence
in-situ
hybridization (FISH) assay
Kalal Iravathy Goud, Seetha Dayakar, Kolanupaka Vijayalaxmi, Saidam Jangu Babu, P Vijay Anand Reddy
March 2012, 135(3):312-317
Background & objectives:
Fluorescence
in situ
hybridization (FISH) is increasingly being recognized as the most accurate and predictive test for H er0 2/
neu
gene amplification and response to therapy in breast cancer. In the present study we investigated HER-2/
neu
gene amplification by FISH in breast carcinoma tissue specimens and compared the results with that of immunohistochemical (IHC) analysis.
Methods:
A total of 90 breast carcinoma tissue samples were used for immunohistochemical (IHC) and FISH analysis. IHC was performed by using mouse monoclonal antibody to the intracellular domain of HER-2/
neu
protein. Each slide was scored in a blinded fashion by two pathologists according to the manufacturer's recommended criteria. FISH analysis was performed on paraffin embedded breast tumour tissue sections. The polysomy for centromere 17 (Spec green signal) was read as green signals less than 4 as moderate polysomy, and more than 4 as highly polysomy.
Results:
Thirty of the 90 patients had negative results by IHC and FISH. Of the 28 patients with the score of 2+ by IHC, 20 were FISH positive for HER-2/
neu
gene amplification, three were FISH negative and five patients showed equivocal (1.8-2.2) results by FISH. These five cases were retested for IHC and FISH on different paraffin embedded tissue blocks, and all five were found positive for HER-2/
neu
gene amplification. Twenty five patients with the score of 3+ by IHC were FISH positive for HER-2/
neu
gene amplification (>2.2). Seven cases with the score of 3+ by IHC were FISH negative for HER-2/
neu
gene amplification (>2.2), and showed polysomy of chromosome number 17 high polysomy > 4.
Interpretation & conclusions:
Our results indicated that HER-2/
neu
status by FISH should be performed in all cases of breast tumour with a 2+ score by IHC. Cases demonstrating a 3+ score by IHC may be subjected to FISH to rule out polysomy of chromosome 17 which could be falsely interpreted as HER-2/
neu
overexpression by IHC analysis. There is also a need for establishing a clinically validated cut-off value for HER-2/
neu
FISH amplification against IHC which may be further compared and calibrated.
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REVIEW ARTICLES
Bone disease in thyrotoxicosis
P Amaresh Reddy, CV Harinarayan, Alok Sachan, V Suresh, G Rajagopal
March 2012, 135(3):277-286
Thyrotoxicosis, a clinical syndrome characterized by manifestations of excess thyroid hormone, is one of the commonly-recognised conditions of the thyroid gland. Thyrotoxicosis causes acceleration of bone remodelling and though it is one of the known risk factors for osteoporosis, the metabolic effects of thyroxine on bone are not well discussed. Studies show that thyroid hormones have effects on bone, both
in vitro
and
in vivo
. Treatment of thyrotoxicosis leads to reversal of bone loss and metabolic alterations, and decreases the fracture risk. There are limited studies in India as to whether these changes are fully reversible. In this review we discuss about the effects of thyrotoxicosis (endogenous and exogenous) on bone and mineral metabolism, effects of subclinical thyrotoxicosis on bone and mineral metabolism and effects of various forms of treatment in improving the bone mineral density in thyrotoxicosis.
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CORRESPONDENCES
Updation of the scale to measure socio-economic status in urban & rural communities in India
SC Tiwari, Ambrish Kumar
March 2012, 135(3):432-434
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ORIGINAL ARTICLES
Phenotypic & molecular characterization of AmpC β-lactamases among
Escherichia
coli,
Klebsiella
spp. &
Enterobacter
spp. from five Indian Medical Centers
Anand Manoharan, Madhan Sugumar, Anil Kumar, Hepzibah Jose, Dilip Mathai, ICMR-ESBL study group
March 2012, 135(3):359-364
PMID
:22561623
Background & objectives:
AmpC β-lactamases which are often plasmid mediated hydrolyze all β-lactam antibiotics except cefepime and carbapenems. We evaluated the presence of AmpC β-lactamases among
Enterobacteriaceae
strains recovered prospectively from patients at five Indian tertiary care centres.
Methods:
The study included 909 consecutive Gram-negative isolates recovered from clinically significant specimens during June 2007 - May 2008 as part of an ICMR-ESBL study. Among the study isolates, 312 were found to be cefoxitin resistant by disc diffusion test (DDT). Minimum inhibitory concentration (MIC) determination by E test was done against amikacin, levofloxacin, impinem, meropenem, ertapenem, tigecycline and piperacillin-tazobactam. Combined DDT using phenyl boronic acid as inhibitor with cefoxitin was used for phenotypic confirmation of AmpC phenotype. The common Amp C genotypes
ACC, FOX, MOX, DHA, CIT
and
EBC
were detected by multiplex PCR.
Results:
Plasmid mediated Amp C phenotype was confirmed in 114 of the 312 (36.5%) cefoxitin resistant isolates with 255 (81.7%) showing multidrug resistance. Susceptibility to tigecycline was highest (99%) followed by imipenem, meropenem (97%), ertapenem (89%), amikacin (85%), and piperacillin-tazobactam (74.6%). Levofloxacin resistance was 82 per cent. ESBL co carriage was observed among 92 per cent of Amp C producers. Among 114 Amp C producers, 48 could be assigned a genotype, this included CIT- FOX (n=25), EBC (n=10), FOX (n = 4), CIT (n=3), EBC-ACC (n=2) and one each of DHA, EBC-DHA, FOX -DHA and FOX-EBC-DHA.
Interpretation & Conclusions:
Overall, AmpC phenotypes were found in 12.5 per cent isolates, multidrug resistance and ESBL co-carriage among them was high suggesting plasmid mediated spread. The study results have implications in rational antimicrobial therapy and continued surveillance of mechanisms of resistance among nosocomial pathogens.
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Retrospective study of chikungunya outbreak in urban areas of India
BN Nagpal, Rekha Saxena, Aruna Srivastava, Neeru Singh, SK Ghosh, SK Sharma, Ashwani Kumar, Hemant Kumar, Alok Suman Sharma, SK Chand, VP Ojha, SS Mohanty, AK Mohanty, RK Dasgupta, G.P.S. Dhillon, AP Dash
March 2012, 135(3):351-358
Background & objectives:
A retrospective study on chikungunya outbreak in India in five States
viz
. Delhi, Madhya Pradesh, Orissa, Maharashtra and Kerala was conducted in 2007-2008 to know the distribution and determinants of chikungunya fever outbreak in India.
Methods:
On the basis of high and low incidence of chikungunya fever, two districts from each State and two wards from the selected district were taken for random selection of 1000 households from 10 districts and 5 States. Semi-structured questionnaires were administered to individuals, patients, qualified health professionals and to stakeholders for collecting information.
Results:
The educational background and occupation of the respondents showed variations across the study States. Only in high incidence ward of Maharashtra, water storage period for 3-6 days and emptying, drying of water containers on weekly basis was noted. The study through knowledge, attitude, belief, practice (KABP) obtained individual's perception of chikungunya fever, its prevention and control. Patients' expenditure on treatment was mainly recorded less than Rs 500 across study States. Health facility survey obtained an overview of the capacity of local health facilities. Stakeholders' perception regarding chikungunya fever was also noted.
Interpretation & Conclusions:
The study revealed differences in awareness of chikungunya, cause of the disease, vector responsible, mode of transmission, biting time and elimination of breeding of mosquitoes statistically significant among high and low incidence wards of all the States. Expenditure on treatment was independent of economically active status and loss of man-days across all the States. Education and occupation did not have any relation with emptying/drying of water containers in high incidence wards. Strengthening of surveillance, information, education and communication (IEC) activities along with case management facilities may be provided by the State health department for prevention of chikungunya outbreaks in future. Stakeholders should be more involved in outbreak management and future planning.
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Antimicrobial resistance in
Shigella
- rapid increase & widening of spectrum in Andaman Islands, India
Debdutta Bhattacharya, AP Sugunan, Haimanti Bhattacharjee, R Thamizhmani, DS Sayi, K Thanasekaran, Sathya Prakash Manimunda, AR Ghosh, AP Bharadwaj, M Singhania, Subarna Roy
March 2012, 135(3):365-370
Background & objectives:
Shigellosis is known to be a major cause of acute childhood diarrhoea in Andaman & Nicobar Islands, India. Rapid emergence of antibiotic resistance warrants continuous monitoring of sensitivity pattern of bacterial isolates. We report here the salient findings of an ongoing study on shigellosis in Andaman Islands, India, with regards to change in drug resistance pattern during the past one decade.
Method:
During 2006-2009, stools samples from 412 paediatric diarrhoea patients were collected and processed for isolation and identification of
Shigella
spp. Susceptibility to 22 antimicrobial drugs was tested and MICs were determined for 3
rd
generation cephalosporins, quinolones, amoxicillin-clavulanic acid combination and gentamicin. Drug susceptibility pattern of these isolates were compared with that of 33 isolates obtained during 2000-2002.
Results:
Shigella
isolates were recovered from 50 of 412 stool samples processed. Resistance to ampicillin, nalidixic acid, tetracycline and ciprofloxacin was observed in 100, 96, 94 and 82 per cent of the isolates, respectively. The frequency of resistance to these drugs was significantly (
P
<0.001) higher than that observed during 2000-2002. Resistance to seven drugs was observed in 2000-2002, whereas resistance to 21 drugs was seen during 2006-2009. The number of drug resistance pattern increased from 13 in 2000-2002 to 43 in 2006-2009. Resistance to newer generation fluoroquinolones, 3
rd
generation cephalosporins and augmentin, which was not observed during 2000-2002, appeared during 2006-2009.
Interpretation & Conclusions:
The frequency of resistance among
Shigella
isolates has increased substantially between 2000-2002 and 2006-2009 and the spectrum of resistance has widened. At present, the option for antimicrobial therapy in shigellosis in Andaman is limited to a small number of drugs. Continuous local monitoring of resistance patterns is necessary for the appropriate selection of empirical antimicrobial therapy.
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Serotyping & molecular characterization for study of genetic diversity among seafood associated nontyphoidal
Salmonella
serovars
Patit Paban Bhowmick, Shabarinath Srikumar, Devananda Devegowda, Malathi Shekar, HA Darshanee Ruwandeepika, Indrani Karunasagar
March 2012, 135(3):371-381
Background & objectives:
Infections due to seafood associated
Salmonella
serovars are great risk to public health. Different phenotypic characteristics have been used previously for epidemiological investigation of
Salmonella
. Beyond the phenotypic characterization, a reliable genetic level discriminatory method is required. Therefore, this study was attempted to use different phenotypic and molecular fingerprinting methods for investigation of genetic diversity among seafood associated nontyphoidal
Salmonella
serovars.
Methods:
Fifty eight seafood associated
Salmonella
isolates were included in this study. All isolates were serotyped and epidemiological investigation was carried out using molecular fingerprinting methods, random amplified polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus sequence based-PCR (ERIC-PCR) along with whole cell protein profiling using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in our study.
Results:
Among the 58
Salmonella
isolates,
S
. Weltevreden was observed to be the most predominant serovar. Typing of
Salmonella
serovars using RAPD and ERIC-PCR suggested the existence of a genetic diversity. Though both PCR based techniques were found to have a good discriminatory index, a better discriminatory ability was observed when the results obtained by the two techniques were combined and taken for composite analysis. Protein profiling of whole cells using SDS-PAGE demonstrated the presence of several bands with two bands of sizes 38 kDa and 46 kDa common among all 58 isolates.
Interpretation & Conclusions:
Our study shows that use of protein profiling in combination with established typing methods such as RAPD and ERIC-PCR may provide useful information in typing of non-typhoidal
Salmonella
isolates associated with seafood and to develop strategies to protect public from
Salmonella
infections.
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CORRESPONDENCES
Dermacentor auratus
Supino, 1897 (Acarina, Ixodidae) reported from Wayanad, Kerala
KG Ajithkumar, Reghu Ravindran, S Ghosh
March 2012, 135(3):435-436
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ORIGINAL ARTICLES
Alleviation of metabolic abnormalities induced by excessive fructose administration in Wistar rats by
Spirulina maxima
Urmila Jarouliya, J Anish Zacharia, Pravin Kumar, PS Bisen, G.B.K.S. Prasad
March 2012, 135(3):422-428
Background & objectives:
Diabetes mellitus is a metabolic disorder characterized by hyperglycaemia. Several natural products have been isolated and identified to restore the complications of diabetes.
Spirulina maxima
is naturally occurring fresh water cyanobacterium, enriched with proteins and essential nutrients. The aim of the study was to determine whether
S. maxima
could serve as a therapeutic agent to correct metabolic abnormalities induced by excessive fructose administration in Wistar rats.
Methods:
Oral administration of 10 per cent fructose solution to Wistar rats (n=5 in each group) for 30 days resulted in hyperglycaemia and hyperlipidaemia. Aqueous suspension of
S. maxima
(5 or 10%) was also administered orally once daily for 30 days. The therapeutic potential of the preparation with reference to metformin (500 mg/kg) was assessed by monitoring various biochemical parameters at 10 day intervals during the course of therapy and at the end of 30 days
S. maxima
administration.
Results:
Significant (
P
<0.001) reductions in blood glucose, lipid profile (triglycerides, cholesterol and LDL, VLDL) and liver function markers (SGPT and SGOT) were recorded along with elevated level of HDL-C at the end of 30 days therapy of 5 or 10 per cent
S. maxima
aquous extract. Co-administration of
S. maxima
extract (5 or 10% aqueous) with 10 per cent fructose solution offered a significant protection against fructose induced metabolic abnormalities in Wistar rats.
Interpretation & Conclusions:
The present findings showed that
S. maxima
exhibited anti-hyperglycaemic, anti-hyperlipidaemic and hepatoprotective activity in rats fed with fructose. Further studies are needed to understand the mechanisms.
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Protective effect of hydroalcoholic extract of
Andrographis paniculata
on ischaemia-reperfusion induced myocardial injury in rats
Shreesh Kumar Ojha, Saurabh Bharti, Sujata Joshi, Santosh Kumari, Dharamvir Singh Arya
March 2012, 135(3):414-421
Background & objectives:
Protecting myocardium from ischaemia-reperfusion (I-R) injury is important to reduce the complication of myocardial infarction (MI) and interventional revascularization procedures. In the present study, the cardioprotective potential of hydroalcoholic extract of
Andrographis paniculata
was evaluated against left anterior descending coronary artery (LADCA) ligation-induced I-R injury of myocardium in rats.
Methods:
MI was induced in rats by LADCA ligation for 45 min followed by reperfusion for 60 min. The rats were divided into five experimental groups
viz.
, sham (saline treated, but LADCA was not ligated), I-R control (saline treated + I-R), benazepril (30 mg/kg + I-R),
A. paniculata
(200 mg/kg
per se
) and
A. paniculata
(200 mg/kg + I-R).
A. paniculata
was administered orally for 31 days. On day 31, rats were subjected to the I-R and cardiac function parameters were recorded. Further, rats were sacrificed and heart was excised for biochemical and histopathological studies.
Results:
In I-R control group, LADCA ligation resulted in significant cardiac dysfunction evidenced by reduced haemodynamic parameters; mean arterial pressure (MAP) and heart rate (HR). The left ventricular contractile function was also altered. In I-R control group, I-R caused decline in superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and reduced glutathione (GSH) as well as leakage of myocytes injury marker enzymes, creatine phosphokinase-MB (CK-MB) isoenzyme and lactate dehydrogenase (LDH), and enhanced lipid peroxidation product, malonaldialdehyde (MDA). However, rats pretreated with
A. paniculata
200 mg/kg showed favourable modulation of haemodynamic and left ventricular contractile function parameters, restoration of the myocardial antioxidants and prevention of depletion of myocytes injury marker enzymes along with inhibition of lipid peroxidation. Histopathological observations confirmed the protective effects of
A. paniculata
. The cardioprotective effects of
A. paniculata
were found comparable to that of benazepril treatment.
Interpretation & Conclusions:
Our results showed the cardioprotective effects of
A. paniculata
against I-R injury likely result from the suppression of oxidative stress and preserved histoarchitecture of myofibrils along with improved haemodynamic and ventricular functions.
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Differential response to sustained stimulation by hCG & LH on goat ovarian granulosa cells
Chanchal Gupta, Trimbak Chapekar, Yash Chhabra, Prashant Singh, Subrata Sinha, Kalpana Luthra
March 2012, 135(3):331-340
PMID
:22561619
Background & objectives:
Chapekar established a model of ovarian tumourigenesis in mice by splenic transplantation of ovaries, resulting in sustained luteinizing hormone (LH) levels because of absence of feedback inhibition. There is increasing evidence of the differential response to LH or hCG under various experimental conditions. The effect of sustained hormonal stimulation in long term cultures is sparsely investigated. The study is aimed to determine the role of hCG and LH stress on caprine ovarian granulosa cells and their downstream signaling in short and long term cultures.
Methods:
To study the response of hCG and LH stress and downstream signaling, short term cultures were set up by exposing goat ovarian granulosa cells in primary cultures to hCG and LH stress (levels beyond their physiological doses) for 5 days (P0). Cells were sub-cultured at sixth day and subjected to prolonged LH/ hCG stress for two weeks in passage 1(P1) (long term cultures). Downstream cell signaling molecules were assessed. Intracellular cAMP was estimated by ELISA. For PKA and PKC, activity assays were performed. pERK protein expressions in short term cultures were assessed by Western blot and fowcytometry; in long term cultures, pERK expression was analyzed by fowcytometry.
Results:
Differential effects on cell proliferation were observed in long term cultures, where the untreated and hCG exposed cells showed markedly reduced cell proliferation after second week of exposure while LH treated cells continued to proliferate. Different levels of cAMP, PKA, PKC and phosphorylated ERK1/2 were observed on short term and long term LH stimulation. On sustained hormonal stimulation, cAMP levels were signifcantly (
P
<0.05) higher in hCG treated cultures as compared to controls and LH treated cultures. LH led to maximal elevation of ERK in long term cultures.
Interpretation & conclusions:
As pERK1/2 promotes cellular proliferation, activation of ERK1/2 in LH treated cultures may be responsible for sustained growth. Prolonged LH treatment promoted growth and proliferation in caprine ovarian granulosa cells whereas prolonged exposure to hCG led to elevated levels of cAMP and decreased the rate of proliferation. Defning the signals and second messengers that act as survival or apoptotic mediators may help in elucidation of the mechanisms controlling proliferation or programmed cell death in granulosa cells.
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Impact of changing over of insecticide from synthetic pyrethroids to DDT for indoor residual spray in a malaria endemic area of Orissa, India
Surya K Sharma, Ashok K Upadhyay, Mohammed A Haque, Prajesh K Tyagi, Bikrant K Kindo
March 2012, 135(3):382-388
Background & objectives:
Development of insecticide resistance in malaria vectors has been a major problem for achieving effective vector control. Due to limited availability of insecticides, the only option is management of resistance by judiciously using the insecticides and rotating them to maintain their effectiveness. This study was carried out in a malaria endemic area of Sundergarh district in Orissa where synthetic pyrethroids (SP) were in use for the last couple of years. The change-over from SP to DDT was done in one arm of study, and the other two arms remained on SP and insecticide-treated nets (ITN). Entomological and parasitological monitoring was done to assess the impact.
Methods:
The study design comprised of three arms
(i)
two rounds of indoor residual spraying (IRS) with DDT 1g/m
[2]
as a change-over insecticide in areas previously under synthetic pyrethroids;
(ii)
two rounds of IRS with synthetic pyrethroid (alphacypermethrin, ACM) @ 25 mg/m
[2]
; and
(iii)
an unsprayed area under ITN/long lasting insecticide nets (LNs). Indoor residual spraying was undertaken under strict supervision to maintain quality and coverage. Contact bioassays were conducted to know the persistence of insecticide on sprayed surfaces and adult vector density was monitored in fixed and randomly selected houses. Malaria incidence was measured through fortnightly domiciliary surveillance under primary health care system in all the study villages.
Results:
The insecticide susceptibility tests showed that
An.culicifacies
was resistant to DDT but susceptible to malathion and ACM. However,
An. fluviatilis
was susceptible to all the three insecticides. ACM was effective in killing
An. culicifacies
on mud and wooden sprayed surfaces and maintained effective bioefficacy ranging from 92 to 100 per cent up to five months, whereas DDT failed to achieve effective mortality in
An.culicifacies
. However, there was significant decline in the density of
An.culicifacies
in ACM and DDT areas in comparison to ITNs/LNs. There was 61 per cent reduction in the slide positivity rate in ACM area in comparison to 48 and 51 per cent in DDT and ITN/LNs areas, respectively. The adjusted incidence rate of malaria cases per 1000 population in three study areas also showed significant declines within each group.
Interpretation & Conclusions:
The present findings show that the change-over of insecticide from synthetic pyrethroids to DDT brings about the same epidemiological impact as envisaged from continuing SP spray or distributing insecticide treated nets/long-lasting insecticidal nets provided there is a good quality spray and house coverage.
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Proteomic profiling of nephrotic syndrome in serum using magnetic bead based sample fractionation & MALDI-TOF MS
Weiguo Sui, Yong Dai, Yue Zhang, Jiejing Chen, Huaying Liu, He Huang
March 2012, 135(3):305-311
Background & objectives:
At present, the diagnosis of nephrotic syndrome (NS) requires a renal biopsy which is an invasive procedure. We undertook this pilot study to develop an alternative method and potential new biomarkers for diagnosis, and validated a set of well-integrated tools called ClinProt to investigate serum petidome in NS patients.
Methods:
The fasting blood samples from 49 patients diagnosed with NS by renal biopsy, including 17 mesangial proliferative glomerulonephritis (MsPGN), 12 minimal change nephrotic syndrome (MCNS), 10 focal segmental glomerulosclerosis (FSGS) and 10 membranous nephropathy (MN), were collected and screened to describe their variability of the serum peptidome. The results in NS group were compared with those in 10 control healthy individuals. Specimens were purified with magnetic beads-based weak cation exchange chromatography and analyzed in a MALDI-TOF MS.
Results:
The results showed 43, 61, 45 and 19 differential peptide peaks in MsPGN, MCNS, MN and FSGS groups, respectively. A Genetic Algorithm was used to set up the classification models. Cross validation of healthy controls from MsPGN, MCNS, MN and FSGS was 96.18, 100, 98.53 and 94.12 per cent, respectively. The recognition capabilities were 100 per cent.
Interpretation & conclusions:
Our results showed that proteomic analysis of serum with MALDI-TOF MS is a fast and reproducible approach, which may give an early idea of the pathology of nephrotic syndrome.
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1,190
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Antihypercholesterolaemic influence of dietary tender cluster beans (
Cyamopsis tetragonoloba
) in cholesterol fed rats
S Pande, K Platel, K Srinivasan
March 2012, 135(3):401-406
Background & objectives:
Cluster beans (
Cyamopsis tetragonoloba
) are rich source of soluble fibre content and are known for their cholesterol lowering effect. The beneficial anti-hypercholesterolaemic effect of whole dietary cluster beans as a source of dietary fibre was evaluated in high cholesterol diet induced hypercholesterolaemia in experimental rats.
Methods:
Male Wistar rats (90-95 g) divided in six groups of 10 rats each were used. Freeze dried tender cluster beans were included at 12.5 and 25 per cent levels in the diet of animals maintained for 8 wk either on high (0.5%) cholesterol diet or basal control diet.
Results:
Significant anti-hypercholesterolaemic effect was seen in cluster bean fed animals, the decrease in serum cholesterol being particularly in the LDL associated fraction. There was also a beneficial increase in HDL associated cholesterol fraction. Hepatic lipid profile showed a significant decrease in both cholesterol and triglycerides as a result of feeding tender cluster beans along with high cholesterol diet.
Interpretation & Conclusions:
The present experimental results showed the beneficial hypocholesterolaemic and hypolipidimic influences dietary tender cluster beans in atherogenic situation. Studies in human need to be done to confirm the results.
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CORRESPONDENCES
Current status of leprosy as seen in a tertiary care hospital in north India
Amita Jain
March 2012, 135(3):439-440
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1,117
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ORIGINAL ARTICLES
Ameliorative effects of
Panax quinquefolium
on experimentally induced reflux oesophagitis in rats
Pratibha Singh, Neetu Singh, Shibani Sengupta, Gautam Palit
March 2012, 135(3):407-413
Background & objectives:
Reflux oesophagitis (RE), is one of the most prevalent chronic gastrointestinal disorders commonly referred to as gastroesophageal reflux disease (GERD) and requires long term therapy. The present study was designed to investigate the protective effects of
Panax quinquefolium (PQ)
, administered with variable doses, on experimentally induced reflux oesophagitis (RE) in rats.
Methods:
Forty two female Sprague-Dawley (180-220 g) rats were randomly divided to receive standardized root powder of PQ (50-200mg/kg, po), standard anti-reflux (omeprazole, 5 mg/kg, ip) and anti-oxidant (α-tocopherol, 16 mg/kg, po). After 45 min drug pretreatment, RE was produced in rats by simultaneous ligation of the pyloric end and forestomach. Several parameters, including macroscopic lesion index, glutathione system, lipid peroxidation (LPO) and tissue myeloperoxidase (MPO) activity were measured. Alterations in ICAM-1, CINC-2 and MCP-1 gene expression were examined through reverse transcriptase polymerase chain reaction (RT-PCR).
Results:
PQ significantly attenuated the severity of the macroscopic signs of RE-induced tissue damage, replenished the depleted GSH level and reduced the RE-associated LPO levels dose dependently. In contrast, omeprazole though effectively improved the mucosal damage, it failed to bring significant attenuation of RE-associated changes in LPO, GSH level and MPO activity. α-Tocopherol significantly ameliorated RE-induced tissue injury and improved LPO level and GSH/GSSG ratio but failed to counteract RE-induced MPO activity. PQ at dose of 100 mg/kg significantly downregulated ICAM-1 and CINC-2 expression whereas it showed no effect over MCP-1 expression.
Interpretation & Conclusions:
The present data indicate that PQ protects against RE-induced oesophageal damage via a mechanism that inhibits the influx of inflammatory cell to oesophagus and a consequence excessive oxidative load, opening the avenue to its promising protective role in patients with gastroesophageal reflux disease (GERD).
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224
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SXT constin among
Vibrio cholerae
isolates from a tertiary care hospital
Kavita Pande, Deepak Kumar Mendiratta, Deotale Vijayashri, Dipak C Thamke, Pratibha Narang
March 2012, 135(3):346-350
Background & objectives:
The SXT element, also known as 'constin' (conjugable, self transmissible, integrating element) is an integrating conjugative element (ICE) in
Vibrio cholerae
discovered in the chromosome of epidemic
V. cholerae
O139 strain MO10 (SXT
MO10
) which arose in late 1992 in Chennai, India. SXT related ICEs have become widespread and currently, most if not all Asian
V. cholerae
clinical isolates contain SXT related ICEs. The present study attempts to determine the presence of SXT
Int
gene in
V. cholerae
recovered between 2005 to 2007 in a tertiary care hospital, demonstrate its conjugal nature and also detect co-presence and co-transfer of plasmids in representative isolates.
Methods:
This prospective study was done on 116
V. cholerae
isolates [114- O1 (107 ogawa and 7 inaba) and 2 - Non O1 Non O139
V. cholerae
] from watery stools between 2005 to 2007 recovered from equal number of patients. PCR was carried out using SXT
Int
specific primers that produced a 592 bp internal fragment of SXT element, and rifampicin resistant strain of
E.coli
K-12 was used as recipient in conjugation experiments to study transfer of SXT, as also co-transfer of resistance to tetracycline, erythromycin, and nalidixic acid. Antibiotic susceptibility was performed against various antibiotics.
Results:
Of the 116 isolates, 110 (94.8%) were positive for SXT element by PCR. It was demonstrated in 94.7 per cent of the O1, and 100 per cent of non O1 non O139
V. cholerae.
All 2005 isolates, 25 per cent of 2006 isolates and 96.6 per cent of 2007 isolates were positive for SXT. Thirty two drug resistance patterns were observed and the 2007 isolates showed resistance to as many as eight antibiotics. The resistance of SXT positive isolates was higher than those of SXT negative and the typical drug resistance pattern corresponding to SXT
ET
and SXT
MO10
was shown by only one
V. cholerae
O1 isolate. Successful conjugal transfer of SXT was seen in 31 (88.6%) of the 35 isolates studied without any co-transfer while, presence of plasmids was observed in two of the 31 donor
V. cholerae
studied.
Interpretation & Conclusions:
The demonstration of SXT element and its successful horizontal transfer in
V. cholerae
isolates studied emphasizes the need for its detection to monitor antibiotic resistance and dissemination in
V. cholerae.
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1,042
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COMMENTARY
Serum proteomics for the diagnosis of nephrotic syndrome: Is there a ray of hope?
Dipankar Bhowmik, Sanjay K Agarwal
March 2012, 135(3):273-275
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1,038
232
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EDITORIALS
India's contribution to global TB control: Innovative & integrated implementation research
Peter M Small, VM Katoch
March 2012, 135(3):267-269
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952
302
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ORIGINAL ARTICLES
Apo E genotyping from blood stored on filter paper
Rizwana Quraishi, Ramakrishnan Lakshmy, Kalpana Luthra, Ashok K Mukhopadhyay, Bansi L Jailkhani
March 2012, 135(3):318-321
Background & objectives:
Dried blood spotted on to filter paper has been found suitable for a large number of studies. In tropical countries with varying temperature conditions the use of dried blood needs to be validated. We carried out this study to assess the use of blood spotted filter paper as a transport system to study genotyping of Apo E gene.
Methods:
Fifty five patients visiting Cardiothoracic Neuroscience Centre (CNC) OPD at the All India Institute of Medical Sciences (AIIMS), New Delhi, and referred for lipid investigations to Cardiac Biochemistry Laboratory were selected at random. Blood was spotted on to Whatman 3 MM filter paper, dried and stored at room temperature. Genomic DNA was extracted and genotyping was carried out at the end of 0, 3 and 12 months. The study was further validated using samples collected on to filter paper from four centres and stored for eight years at room temperature. The temperature and humidity conditions of the centre varied widely.
Results:
Fifty five samples collected on to filter paper showed exact match of the genotyping when compared to fresh blood. In dried blood samples collected and stored for 1 yr at room temperature DNA extraction and apo E genotyping was done successfully.
Interpretation & conclusions:
The present results showed the feasibility of using dried blood samples on filter paper for apo E genotyping in tropical temperature. The findings need to be validated on a large sample before being recommended for use.
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CORRESPONDENCES
JAK2
mutation in patients with splanchnic venous thrombosis: A pilot study from India
S Sazawal, S Rathi, S Chikkara, R Chaubey, T Seth, A Saraya, J Das, M Mahapatra, R Saxena
March 2012, 135(3):429-431
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ORIGINAL ARTICLES
Evaluation of toxicity of plant extracts against vector of lymphatic filariasis,
Culex quinquefasciatus
M Sakthivadivel, Alex Eapen, AP Dash
March 2012, 135(3):397-400
Background & objectives:
Conventional insecticides are generally used as larvicides to control
Culex quinquefasciatus
, vector of lymphatic filariasis. This study was undertaken to evaluate the larvicidal activity of some potential larvicidal plants leaf extracts against
Cx. quinquefasciatus
larvae.
Methods:
The toxic effects of petroleum ether leaf extracts of plants
viz
.,
Argemone mexicana
(Mexican prickly poppy)
, Clausena dentata
(Dentate)
, Cipadessa baccifera
(Rana bili)
, Dodonaea angustifolia
(Hop bush) and
Melia dubia
(Pride of India) were evaluated under laboratory conditions in individual and in combination against 3
rd
- 4
th
instar larvae of
Cx. quinquefasciatus
.
Results:
The results indicated that among the selected plants,
A. mexicana
showed maximum larvicidal activity with an LC
50
value of 48.89 ppm. Its toxicity was enhanced when the extract was mixed (1:1) with that of
C. dentata
as the LC
50
value became 28.60 ppm indicating synergistic action of
A. mexicana.
Interpretation & Conclusions:
Our results showed high larvicidal potential in
A. mexicana
leaf extract, and it also showed additive effect when mixed with
C. dentata
extract.
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EDITORIALS
Defining & counting malaria deaths
Naman K Shah
March 2012, 135(3):270-272
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919
206
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CORRESPONDENCES
Vancomycin resistance among MRSA
V Anil Kumar
March 2012, 135(3):441-441
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779
250
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Cases of pandemic influenza A (H1N1) 2009 in western Rajasthan, India
Vinod Joshi, Bennet Angel, Anil Purohit, Himmat Singh, Neetu Bohra, Annette Angel, Rashmi Chauhan, Manju Singhi, Arvind Mathur
March 2012, 135(3):437-438
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844
168
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BOOK REVIEWS
Evaluation of certain food additives and contaminants, Seventy-third report of the Joint FAO/WHO Expert Committee on Food Additives
AK Sharma
March 2012, 135(3):446-447
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818
181
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Recombinant antibodies for immunotherapy
NK Mehra, Prashant Sood
March 2012, 135(3):444-445
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779
184
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CORRESPONDENCES
Authors' response
V Thati, CT Shivannavar, SM Gaddad
March 2012, 135(3):441-442
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574
145
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BOOK REVIEWS
The elite young athlete
Jaspal Singh Sandhu
March 2012, 135(3):443-443
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532
157
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BOOKS RECEIVED
Book Received
March 2012, 135(3):447-447
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Online since 25 February, 2011