Entanglement of UPRER in Aging Driven Neurodegenerative Diseases.

The endoplasmic reticulum (ER) is an indispensable cellular organelle that remains highly active in neuronal cells. The ER bears the load of maintaining protein homeostasis in the cellular network by managing the folding of incoming nascent peptides; however, the stress imposed by physiological/environmental factors can cause ER dysfunctions that lead to the activation of ER unfolded protein response (UPRER). Aging leads to deterioration of several cellular pathways and therefore weakening of the UPRER. The decline in functioning of the UPRER during aging results in accumulation of misfolded proteins that becomes intracellular inclusions in neuronal cells, resulting in toxicity manifested as neurodegenerative diseases. With ascension in cases of neurodegenerative diseases, understanding the enigma behind aging driven UPRER dysfunction may lead to possible treatments.

Implications of aging and the endoplasmic reticulum unfolded protein response on the molecular modality of breast cancer

The endoplasmic reticulum (ER) is an important subcellular organelle that is involved in numerous activities required to achieve and maintain functional proteins in addition to its role in the biosynthesis of lipids and as a repository of intracellular Ca²⁺. The inability of the ER to cope with protein folding beyond its capacity causes disturbances that evoke ER stress. Cells possess molecular mechanisms aimed at clearing unwanted cargo from the ER lumen as an adaptive response, but failing to do so navigates the system towards cell death. This systemic approach is called the unfolded protein response. Aging insults cells through various perturbations in homeostasis that involve curtailing ER function by mitigating the expression of its resident chaperones and enzymes. Here the unfolded protein response (UPR) cannot protect the cell due to the weakening of its protective arm, which exacerbates imbalanced homeostasis. Aging predisposed breast malignancy activates the UPR, but tumor cells maneuver the mechanistic details of the UPR, favoring tumorigenesis and thereby eliciting a treacherous condition. Tumor cells exploit UPR pathways via crosstalk involving various signaling cascades that usher tumor cells to immortality. This review aims to present a collection of data that can delineate the missing links of molecular signatures between aging and breast cancer.

Extended spectrum beta-lactamases in Escherichia coli & Klebsiella pneumoniae & associated risk factors.

BACKGROUND & OBJECTIVE: Extended spectrum beta-lactamases (ESBLs) have emerged as a major threat worldwide with limited treatment options. The genotypes of ESBL producing strains largely remain unknown in India; hence the present study was aimed to determine the occurrence of ESBLs in Escherichia coli and Klebsiella pneumoniae, their molecular types and associated risk factors in a tertiary care hospital. METHODS: Total 200 consecutive clinical isolates of E. coli (n=143) and K. pneumoniae (n=57) collected between February and July 2006 at Sanjay Gandhi Postgraduate Institute of Medical Sciences, a tertiary care hospital in north India, were examined phenotypically for ESBL production. ESBL strains were further typed for the bla(TEM/SHV/CTX-M) genes by PCR using specific primers. The bla(CTX-M) cluster was identified by restriction analysis and genotype by sequencing of PCR product. Resistance to other antimicrobial agents was also studied. Various risk factors associated with ESBL infections were analyzed by logistic regressions. RESULTS: ESBLs were found in 63.6 per cent E. coli and 66.7 per cent K. pneumoniae isolates. Majority of the typeable isolates harboured two or more ESBL genes (57.3%). Overall bla(CTX-M) was the commonest genotype (85.4%) followed by bla(TEM) (54.9%) and bla(SHV) (32.9%) either alone or in combination. All CTX-M enzymes in E. coli and 87.5 per cent in K. pneumoniae belonged to the CTX-M-1 cluster. Sequencing was done for randomly selected 20 bla(CTX-M) PCR products and all were identified as CTXM- 3. Resistance of ESBL isolates to other antibiotics was amikacin 14.7 per cent, gentamicin 66.7 per cent, trimethoprim/sulphamethoxazole 79.1 per cent and ciprofloxacin 93.8 per cent. Prior antibiotic exposure, use of intravenous device and urinary catheter, renal insufficiency and ICU admission were associated with ESBL infection on univariate analysis. On multivariate, antibiotic exposure (P=0.001) and use of urinary catheter (P<0.001) were identifified as risk for ESBL infection. INTERPRETATION & CONCLUSION: Our study showed high ESBL occurrence with CTX-M as the emerging type in our hospital and CTX-M-3 being reported for the fi rst time in India. High co-resistance to other non-beta-lactam antibiotics is a major challenge for management of ESBL infections.

Non-polio enteroviruses in acute flaccid paralysis children of India: vital assessment before polio eradication.

AIM: This study is an overview of non-polio enterovirus (NPEV) circulating in North India studied from the perspective of poliomyelitis eradication. Wild polio cases declined because of intensive oral polio vaccine immunization. As we approach global eradication of poliovirus (PV), NPEV causing acute flaccid paralysis (AFP) are equal cause of concern. METHODS: A total of 46 653 AFP samples (World Health Organization) and apparently 1000 healthy contacts living in the same geographical area were studied (2004-2007). Serological identification of NPEV was done using RIVM-specific pools (The Netherlands). Untyped (UT)-NPEVs were sequenced directly from reverse transcription-polymerase chain reaction using pan-enterovirus (Pan-EV) primer (CDC, Atlanta, GA) targeting highly conserved 5'un-translated regions of the enterovirus. RESULTS: In this study, 12 513 NPEVs were isolated from the collected stool samples. Seroneutralization had identified 67% of NPEV isolates, whereas 32.6% remained as UT- NPEV. Of the typed NPEVs, Coxsackie-B accounted for 32.3%; followed by echoviruses-11, 12, 13, 7 between 8 and 28%. In sequencing few UT-NPEVs, some were identified also as echovirus-30, 11 and 18 which were probably present in mixtures as they remained UT-NPEV in ENT. Newly classified human enterovirus virus-86 (HEV) (EU079026), HEV-97(EU071767) and HEV-B isolate (EU071768) were isolated in AFP samples. CONCLUSIONS: This study provided definitive information about circulation, prevalence and new emerging NPEV in the polio-endemic region of India, hence they should be considered in AFP surveillance. This would help in adopting and planning new strategies in post-PV eradication era in the country. This is the right time to prepare for the future tasks while we head towards a polio-free region.

emb nucleotide polymorphisms and the role of embB306 mutations in Mycobacterium tuberculosis resistance to ethambutol.

The emb locus has been considered a target for ethambutol (EMB). Substitutions of codon 306 in Mycobacterium tuberculosis embB have been shown to be the most frequent and predictive mutations for EMB resistance; however, recent reports question the biological role of this mutation. We sequenced embB, embC and embR of 44 EMB-resistant M. tuberculosis strains and found that 30/44 (68.1%) strains had a resistance-associated mutation in one of the three genes sequenced. The majority of these mutations resulted in amino acid replacements at codon 306, 368, 378, and 406 of EmbB. The most common mutation reported in EmbC was at codon 270, followed by mutation at codon 297. Novel mutations were also reported in EmbR. Mutations in embC and embR were usually present together with mutations in embB. We found 41/44 EMB-resistant isolates to be resistant to other antituberculosis drugs as well. Our data confirm that mutation at emb306 does not confer resistance to EMB but is a rather common polymorphism in clinical strains of M. tuberculosis predisposing them to the development of any type of drug resistance.

Frequency of Helicobacter pylori and CagA antibody in patients with gastric neoplasms and controls: the Indian enigma.

BACKGROUND: Despite association between H. pylori and gastric neoplasm (GN) from the developed world, studies from India, where infection is more common and acquired early, are scant and contradictory. METHODS: Two hundred and seventy-nine patients with GN from two northern and one eastern Indian centers during the period 1997-2005, 101 non-ulcer dyspepsia (NUD), and 355 healthy volunteers (HV) were evaluated for H. pylori [rapid urease test (RUT), histology and anti-H. pylori, and CagA IgG serology]. RESULTS: Patients with GN [263 gastric carcinoma and 16 (6%) primary gastric lymphoma, 208 male] were older than HV (n = 355, 188 male) and NUD (n = 101, 54 male) patients (53 +/- 12 versus 44 +/- 17 and 43 +/- 13 years, respectively; P < 0.001). Eastern Indian patients with GN (n = 145) were younger than those from northern India (n = 134; 52 +/- 12 versus 55 +/- 12 years; P < 0.007, t-test). In GN and NUD patients H. pylori positivity by RUT [86/225 (38%) versus 46/101 (46%)], anti-H. pylori IgG [154/198 (78%) versus 85/101 (84%)], and histology [136/213 (64%) versus 55/101 (55%)] were comparable (chi(2)-test). Serum IgG anti-H. pylori antibody was more common among HV than among GN patients [300/355 (85%) versus 154/198 (78%); P = 0.04, chi(2)-test]. Intestinal metaplasia was more common in GN than in NUD patients [101/252 (40%) versus 2/98 (2%), P < 0.000, chi(2)-test]. CagAIgG was more common in GN than in NUD patients [124/163 (76%) versus 64/101 (63%)] but comparable to that in HV patients [87/98 (89%), P = NS]. CONCLUSION: Frequency of H. pylori as detected using endoscopy and serology-based tests is not higher among patients with GN as compared with controls in India.

Emerging vancomycin resistance in enterococci in India.

Infection caused by vancomycin resistant enterococci (VRE) leads to adverse outcome and is a real challenge. Despite increasing reports of VRE in different countries, there is scanty data on this issue from India. A total of 685 enterococci were isolated from various clinical samples from January to December 2004. Antimicrobial susceptibility was performed as prescribed by National Committee for Clinical Laboratory Standards (NCCLS). Vancomycin resistance was confirmed by minimum inhibitory concentration (MIC). Resistant phenotype was determined by Polymerase chain reaction (PCR). Of 685, 456 (67%) were E. faecalis and 229 (33%) were E. faecium. Resistance to various antibiotics in E. faecalis and E. faecium was as follows: ampicillin 33% and 54%, erythromycin 91% and 86%, ciprofloxacin 69% and 81%, tetracycline 50% and 54% and high level gentamicin resistance in 62% and 77% respectively. Vancomycin resistance was confirmed in 10 (1.4%) cases by MIC and all had Van A phenotype by PCR. Emergence of vancomycin resistant enterococci is of great concern because of its epidemic potential and scanty therapeutic options. Prompt diagnosis and efficient infection control measures can restrict its spread.

1H NMR spectroscopy in the diagnosis of Klebsiella pneumoniae-induced urinary tract infection.

The (1)H NMR spectroscopic method is suggested and its utility is demonstrated for the diagnosis of Klebsiella pneumoniae (K. pneumoniae) in urinary tract infection (UTI). K. pneumoniae have the specific property of metabolizing glycerol to 1,3-propanediol (1,3-PD), acetate, ethanol and succinate. The quantity of 1,3-PD produced correlates well with the viable bacterial count. Other common bacteria causing UTI (except for Citrobacter frundii), such as Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Enterobacter aerogenes, Acinetobacter baumanii, Proteus mirabilis, Enterococcus faecalis, Streptococcus gp B and Staphylococcus aureus do not metabolize glycerol under similar conditions. Citrobacter frundii (C. frundii) also gives the same NMR results but is easily differentiated as being motile on direct microscopic examination of urine and it is not common nosocomial infectious agent in urinary tract infection. The method provides a single-step documentation of K. pneumoniae (and C. frundii) qualitatively as well as quantitatively. Out of the total 614 subjects considered, clinical diagnosis of UTI was obtained in 516 cases (84%). The NMR-based screening had a sensitivity of 90%, a specificity of 100% and a false negativity of 10% relative to the conventional quantitative culture method. In the present authors' experience, the results of NMR spectroscopy based screening show a very good correlation with the diagnosis of urinary tract infected patients.

Helicobacter pylori-induced apoptosis in pathogenesis of gastric carcinoma.

BACKGROUND: Despite a possible role of Helicobacter pylori in gastric carcinoma (GC), its pathogenesis is not clear. There is scanty data on apoptosis in GC in relation to H. pylori and CagA antibody. Therefore, we studied gastric epithelial apoptosis in GC and non-ulcer dyspepsia (NUD) with or without H. pylori infection, and the degree of apoptosis in relation to CagA antibody status. METHODS: 20 patients each with GC and NUD were investigated for H. pylori using rapid urease test (RUT), IgG anti-H. pylori and anti-CagA antibodies, histology of endoscopically normal-looking mucosa for H. pylori, intestinal metaplasia (IM), and apoptosis using TUNEL assay. Positivity to one tissue-based (RUT or histology) and one serology based (anti-H. pylori or CagA IgG) test was taken as diagnostic of active H. pylori infection, and negative result in both tissue-based tests suggested its absence. RESULTS: Patients with GC more often had anti-H. pylori IgG (16 of 20 vs. 8 of 20; p=0.02) and a trend towards higher apoptotic index (AI) (48.6 [19.2 to 71.7] vs. 41.4 [11.7 to 63.6]; p=0.06) than NUD. AI was higher in GC (66.7 [57.5 to 71.7] vs. 32.6 [19.2 to 39.8]; p<0.0001) and NUD (58.6 [50.7 to 63.6] vs. 24.4 [11.7 to 32.2]; p<0.0001) infected with H. pylori than in those without infection. AI was also higher in GC than in NUD with H. pylori infection (66.7 [57.5 to 71.7] vs. 58.6 [50.7 to 63.6]; p=0.01). Four of the 20 patients with GC and none with NUD had IM (p=ns). There was no difference in AI in relation to CagA antibody. AI positively correlated with patients' age in presence of H. pylori infection (correlation coefficient=0.5, p=0.03) but not in its absence. CONCLUSION: Exaggerated apoptosis may play a role in H. pylori-mediated gastric diseases including carcinogenesis. AI increases with aging in patients infected with H. pylori.

Nocardial brain abscesses in a HIV positive patient misinterpreted as tubercular brain abscesses.

We present a case of nocardial (Nocarda transvalensis) brain abscesses in a HIV infected person with CD4 count of 53 cells/ml, who received antitubercular therapy for one year. A magnetic resonance imaging study showed multiple ring-enhancing lesions in right parieto-occipital parenchymal region along with perilesional edema and mass effect. Right posterior temporal burr hole aspiration of the abscesses and postoperative cotrimoxazole and ampicillin-sulbactum therapy cured the patient. It is a case of HIV infection with rare and sole manifestation of multiple cerebral abscesses due to N. transvalensis.

(1)H NMR spectroscopy in the diagnosis of Pseudomonas aeruginosa-induced urinary tract infection.

The utility of (1)H NMR spectroscopy is suggested and demonstrated for the diagnosis of Pseudomonas aeruginosa in urinary tract infection (UTI). The specific property of P. aeruginosa of metabolizing nicotinic acid to 6-hydroxynicotinic acid (6-OHNA) is exploited. The quantity of 6-OHNA produced correlates well with the viable bacterial count. Other common bacteria causing UTI such as Escherichia coli, Klebsiella pneumonia, Enterobacter aerogenes, Acinetobacter baumanii, Proteus mirabilis, Citrobacter frundii, Enterococcus faecalis, Streptococcus gp B and Staphylococcus aureus do not metabolize nicotinic acid under similar conditions. The method provides a single-step documentation of P. aeruginosa qualitatively as well as quantitatively. The NMR method is demonstrated on urine samples from 30 patients with UTI caused by P. aeruginosa.

First documented cure of a suggestive exogenous reinfection in polymyositis with same but multidrug resistant M. tuberculosis.

BACKGROUND: MDR Mycobacterium tuberculosis is the major cause of treatment failure in tuberculosis patients, especially in immunosuppressed. We described a young polymyositis patient on immunosuppressive therapy who was started with antituberculosis therapy as a susceptible strain of M. tuberculosis was isolated from a single cutaneous abscess in his neck and from regional lymph nodes. CASE PRESENTATION: He had non-reactive miliary tuberculosis and multiple cutaneous abscesses 6 months later with the same strain, which was resistant this time to 9 antituberculosis drugs. We described clinical presentation, radiological and laboratory work-up, treatment and follow-up as the patient was cured after 1.5 years with 6 antituberculosis drugs. CONCLUSION: To our knowledge, this is the first reported case where an immunosuppressed patient with suggestive exogenous reinfection within 6 months with the same but MDR strain of M. tuberculosis was cured. Intense management and regular follow up were important since the patient was a potent source of MDR M. tuberculosis infection and there was limited choice for therapy.

Infection with cytomegalovirus in patients with inflammatory bowel disease: prevalence, clinical significance and outcome.

Despite frequent use of immunosuppressive drugs in patients with inflammatory bowel disease (IBD) and reports of cytomegalovirus (CMV) infection following post-transplant immunosuppression, data on the frequency and clinical significance of CMV in patients with IBD are scant. Sixty-three patients with IBD (61 ulcerative colitis and two Crohn's disease) were evaluated for CMV using serology (IgM antibody, mu-capture ELISA), PCR for CMV DNA in colonic biopsy and histological assessment of haematoxylin and eosin-stained colonic biopsy. Positive result in any test was considered as CMV infection. Various parameters associated with CMV infection were analysed using univariate and multivariate analysis. Ten of 63 (15.8 %) patients (age 36.0 +/- 11.2 years, 31 female) were infected with CMV (DNA alone in four, IgM antibody alone in two and both in four, inclusion body in one). Patients with CMV infection were more often female (8/10 vs 23/53, P < 0.05), had pancolitis (10/10 vs 33/53, P < 0.05), histological activity (9/10 vs 17/53, P < 0.005) and used azathioprine (5/10 vs 7/53, P = 0.04; Fisher exact test for all). On multivariate analysis, female gender, pancolitis and histological activity were the independent factors associated with infection. Patients with CMV infection more often required surgical treatment for IBD (4/10 vs 4/53, P = 0.01) and had fatal outcome (3/10 vs 0/53, P = 0.003). CMV infection in patients with IBD may be common and is associated with poor outcome. PCR of rectal biopsy was the most sensitive method of detection followed by IgM antibody for diagnosis.

Multisystem involvement of microfilaria in a HIV positive patient.

A 35-year-old HIV positive male presented with dyspnoea and chest pain was diagnosed having acute pericardial and pleural effusion. Microfilaria was detected from blood as well as from the pericardial and pleural fluid and from urine. CD4 count was 123 cells microl. The patient was receiving treatment with antiretroviral therapy and Cotrimoxazole for last 4 months. The patient had no opportunistic infection and no symptoms suggestive of filarial infection in the past. This is for the first time we are reporting high microfilarial load (1000/ml) from blood in HIV positive patient, where microfilaria was also demonstrated from the pericardial and pleural fluid and from urine.

Two novel clinical presentations of Burkholderia cepacia infection.

We report two cases of multidrug-resistant Burkholderia cepacia (B. cepacia genomovar I) and Burkholderia multivorans causing multiple liver abscesses in a patient with bronchial asthma (case 1) and peritonitis in a patient with cirrhosis and hepatitis C virus disease (case 2), respectively. Both patients were treated successfully.

A comparative evaluation of phenotypic and molecular methods for the detection of oxacillin resistance in coagulase-negative staphylococci.

Detection of oxacillin resistance in coagulase-negative staphylococci (C-NS) by phenotypic methods is often difficult. The present study compared the National Committee for Clinical Laboratory Standards (NCCLS) revised guidelines of phenotypic methods with a mecA-based polymerase chain reaction (PCR) for C-NS. Ninety clinical C-NS isolates were tested for oxacillin resistance by disk diffusion (1-microg disk), minimum inhibitory concentration (MIC) breakpoint (0.5 microg/ml) after 24 h, and mecA-based PCR. The sensitivity and specificity of disk diffusion was 80% and 93%, and the sensitivity and specificity of the MIC breakpoint after 24 h was 84% and 91%, respectively, against PCR as gold standard. Eleven strains (7 mecA-positive and 4 mecA-negative) showed discordant results between MIC breakpoint after 24 h and PCR. Six of the 7 mecA-positive and all 4 mecA-negative discordant strains had inducible oxacillin resistance and beta-lactamase hyperproduction, respectively. The present study concludes that inducible oxacillin resistance and beta-lactamase hyperproduction are the major causes of discordant results between phenotypic methods and mecA-based PCR, and need special attention.

Frequency of leptospirosis in patients of acute febrile illness in Uttar Pradesh.

OBJECTIVE: This study was performed to asses the disease burden of leptospirosis in and around Lucknow among patients presenting with acute febrile illness and conforming to the case definition of leptospirosis. METHODS: A total of 346 serum samples (mostly paired) and an equal number of urine samples were collected from patients presenting with acute febrile illness and fulfilling the criteria of clinical diagnosis of leptospirosis from January 2001 to December 2001. Patients attending a tertiary care hospital as well as from various communities in and around Lucknow were included in this study. All sera and urine samples were tested for the presence of IgM antibody by Leptodipstick test and by dark-field microscopy (DFM) respectively. All positive and 10% negative sera were tested at national leptospirosis reference centre at Andaman and Nicobar Islands for microagglutination test (MAT). RESULTS: IgM antibody was detected in 25/346 (7%) patients ranging in age from 9-65 years. DFM was positive in only in one case. MAT was positive in 4/17 cases tested and the prevalent serogroups were L. grippotyphosa and L. pomona in two each. Common presenting features in these patients were fever (25/25) and jaundice (17/25). History of contact with animal or water contaminated with animal urine was present in 96% cases. CONCLUSION: Leptospirosis is not uncommon in Uttar Pradesh. However larger epidemiological studies are required to know the actual disease burden. Dark-field microscopy is an insensitive method for the diagnosis of leptospirosis and is not suitable for surveillance.

Neurocysticercosis in clinically suspected and MRI proven cases: evidence of sub-optimal antibody response.

Neurocysticercosis (NCC) has a worldwide distribution mainly in the developing countries like India. The study was done to find the seroprevalence of anti-cysticercus antibodies in clinically suspected and MRI proven cases and to corroborate the serological findings with radiological findings (MRI). A hospital based study among 204 suspected patients during January, 1996 to August, 2001 showed that 77 (32.2%, M:F = 2.2:1) had serological evidence of NCC. Of the total 189 sera, tested at 1:100 dilution 68 (35.9%) and of the total 50 CSF, tested at 1:5 dilution 9 (18%) were positive for anti-cysticercus IgG antibodies. In 35 cases where both were tested 13 sera (37.1%), 9 CSF (25.7%) and in 7 (20%) both sera and CSF were positive. In CSF from 62 patients with tubercular meningitis (disease control) 2 (3.2%) samples whereas in sera of 60 normal blood donors (normal control) 7 (11.7%) samples had anti-cysticercus IgG antibodies. In 33 MRI-positive cases, anti-cysticercus antibodies were seen in 15 (45.4%) patients. Antibodies were seen in 6 of 14 (42.8%) cases with single cortical cyst, 4 of 11 (36.3%) with 2-3 cysts and in 5 of 8 (62.5%) with multiple cysts. Alternatively, 18 of 33 (54.5%) MRI positive cases lacked anti-cysticercus antibodies. Six MRI negative cases were found to be seropositive and were treated successfully. Hence, immune response was sub-optimal even in MRI positive cases and conversely, few MRI negative cases were seropositive. Since positive response with MRI or serology depends on the stage of the disease, therefore both tests should be done together to confirm or to rule out NCC.

Sero-molecular evaluation of human cytomegalovirus disease in renal transplant rejection.

Cytomegalovirus (CMV) is the most common viral pathogen in renal transplant recipients resulting in graft rejection. The prevalence of CMV disease and renal graft rejection is not well studied in India. Sequential specimens from 32 renal allograft recipients were examined by using CMV IgM specific mu capture ELISA and DNA by PCR. Twelve of the 32 patients were CMV IgM positive and out of 12 patients, 9 had rejection and 4 experienced CMV disease. CMV IgM specific mu capture ELISA helped in diagnosis of CMV disease, though it is less sensitive in detection of rejection. PCR itself was proved not sensitive enough in detecting either CMV disease or rejection. At present, optimal laboratory detection of CMV infection in these patients can be achieved only by multiple and more sensitive parameters.