Assessment of Knowledge and Practice of Blood Transfusion Among Nurses in a Tertiary Care Hospital in India.

The purpose of this study was to assess the knowledge and awareness of blood transfusion practices among nurses working in a tertiary care hospital. The objective was to make use of the results to decide the necessity of targeted teaching using lectures and simulated ward scenes. This was a cross sectional study in which a questionnaire comprising of 25 single best-response type multiple choice questions related to blood products and blood transfusion was distributed to nurses who were selected randomly. Questions were both knowledge and practice based. Five hundred and forty-six nurses consented and were assessed. The data was collected, entered and statistically assessed. The number of 'Correct', 'Incorrect' and 'Don't Know' answers were noted. Each correct answer was awarded 1 point, whereas a wrong answer and a 'Don't Know' answer received no points. The individual scores were noted and then multiplied by 4 to get a percentage value. Nurses with 1-5 years of experience scored statistically better than nurses with < 1 year and > 5 years of experience. Nurses working in the haematology-oncology ward scored the most number of correct responses, followed by nurses working in ICU. Only 9.9% of nurses answered > 80% questions correctly. Nurses who had 1-5 years of experience scored better. All nurses were trained in blood transfusion at induction. Though there were occasional non-compulsory lectures as ongoing programs, they had no specific impact on knowledge and awareness. The authors suggest that targeted and regular simulated training is essential at all levels of nursing experience.

Internalisation of hepatitis C virus core protein by human conjunctival fibroblasts.

Recent studies indicate that hepatitis C virus (HCV) proteins can mediate innate immune response and inflammation in conjunctival fibroblasts which contributes to the pathology of dry eye condition associated with chronic HCV infection. The present study investigates the phagocytic potential of human conjunctival fibroblasts (HCFj) for HCV core protein. HCFj cells were incubated with HCV core antigen for different periods of time, and fluorescent micrographs were taken to observe protein internalisation. HCFj cells were capable of internalising HCV core antigen within 1 h; this gives an insight into another molecular mechanism which may contribute towards HCV-associated conjunctival inflammation.

Serum-derived hepatitis C virus 1a infection of human astrocyte cell line SVG.

Neuroinvasion of hepatitis C virus (HCV) is evidenced by recent clinical studies. In this study, serum-derived HCV infection of astrocytes was analysed. Astrocytes were infected with HCV-positive serum, and viral replication was assessed on different days postinfection. RT-PCR was positive for HCV-negative strand on 5th and 7th day postinfection in the HCV-positive serum-infected astrocytes. Real-time RNA count in the cell culture supernatant was steadily increasing from day 3 to day 7. To reconfirm the viral replication, astrocytes were treated with an antiviral before the serum infection, and the antiviral treatment significantly reduced the viral RNA count. Further, the virus-infected cells stained positive for the presence of viral core protein. Electron microscopy revealed the presence of HCV-like particles in the astrocyte cell culture supernatant. In conclusion, serum-derived HCV replicates in human astrocyte cell line SVG.

Mebiolgel, a thermoreversible polymer as a scaffold for three dimensional culture of Huh7 cell line with improved hepatocyte differentiation marker expression and HCV replication.

PURPOSE: A novel three dimensional (3D) culture system purely synthesised from co-polymer which is free from biological contamination for Huh7 cell cultivation and hepatitis C virus (HCV) replication has been attempted. MATERIALS AND METHODS: Mebiolgel, a thermo-reversible gelation polymer was used as a 3D scaffold for culturing Huh7, a liver carcinoma cell line used in our study. The 3D culture of the cells were infected with cell culture derived HCV. RESULT: The scaffold supported the cell growth as 3D spheroids for up to 63 days. Moreover mebiolgel was found to be improving the hepatocyte differentiation of Huh7 cells at the transcript level. Three dimensional culture was susceptible for HCV infection, and this was confirmed by detecting the HCV replication intermediate viral core antigen. CONCLUSION: Mebiolgel based culture system was proven to be suited for 3D culture of Huh7 cells by improvising liver specific genotypic expression and was susceptible for HCV replication. Since mebiolgel based Huh 7 express better hepatocyte differentiation markers genotypically, this can be implemented as an alternate for primary hepatocytes in studies such as viral isolation from patient serum.

Optimisation and analysis of polymerase chain reaction based DNA sequencing for genotyping polyoma virus in renal transplant patients: a report from South India.

PURPOSE: To optimise a polymerase chain reaction (PCR) based DNA sequencing technique for genotyping polyoma virus in clinical specimens obtained from renal transplant patients. MATERIALS AND METHODS: A hundred and thirty (106 peripheral blood and 24 urine) clinical specimens collected from renal transplant patients were included in the study for detecting the presence of  DNA of BK virus (BKV), JC virus (JCV) by PCR targeting the viral protein 1 (VP1) gene. PCR based DNA sequencing was performed to determine the genotypes of polyoma virus and subjected to bioinformatics analysis to determine the amino acid sequences and screen for mutations in the VP1 gene. RESULTS: Polyoma virus was detected in 23 (17.69%) specimens of which 19 (82.60%) were positive for BK virus, 3 (13.04%) for JC virus and 1 for both BK and JC virus. PCR based DNA sequencing detected BK virus genotype I in 12 (50%), genotype IV in 8 (33.3%) and JC virus in 4 (16.6%) clinical specimens. BKV genotype I was the predominant genotype (64.2% in peripheral blood and 33.33% in urine) prevalent in south India. Six novel mutations were found--at position 29, 30 to 47 of BKV genotype I; at position 11 and 15 of BKV genotype IV and at position 2 and 30 of JCV. CONCLUSION: BKV genotype I is the prominent genotype in India and novel mutations detected in the VP1 gene of BKV and JCV are being reported for the first time in literature.

Indigenously produced polyhydroxyalkanoate based co-polymer as cellular supportive biomaterial.

The focus of this study was to evaluate the growth of the cells on a scaffold based on novel polyhydroxyalkanoate (PHA) (Polyhydroxy propionate copoly hydroxy ocatadecanoate copolymer), derived from a mutant strain of Pseudomonas sp. Naive PHA was also blended with several biodegradable polymeric materials (PEG, PLA, and MMT) to improve the scaffold properties. Protein adsorption study was done to evaluate the capability of scaffolds for cellular interaction. PHA:PEG blended scaffold showed better adsorption than others. 3T3 fibroblast cultures on various polymers were equally viable when compared with control culture except for the blend PHA:MMT by CCK 8 kit. MTT assay, performed with the continuous cultures HeLa, HEp-2, Vero, and McCoy on the polymer blends, supported the above finding. Among the blends PHA:PEG showed increased viability and was selected for further studies. Cell proliferation assay with colorimetric BrdU ELISA kit showed increase in cell proliferation over the matrix PHA:PEG than that of control. There were no observable morphological changes of continuous cells grown over matrix PHA:PEG when observed by phase contrast microscopy. HEp-2 cells were enclosed within the matrix when analyzed by SEM. The current study states that the scaffold prepared by using the indigenous PHA in combination with PEG supports cell growth better than the conventional plastic surface. PHA:PEG would be a promising material for tissue engineering.

Clonal diversity of New Delhi metallobetalactamase-1 producing Enterobacteriaceae in a tertiary care centre.

PURPOSE: New Delhi metallobetalactamase-1 (NDM-1) production is a major mechanism of resistance to carbapenems among the Enterobacteriaceae and is a cause for concern in the field of microbial drug resistance. This study was performed to detect NDM-1 in Enterobacteriaceae and to determine the clonal relatedness of NDM-1 producing Escherichia coli and Klebsiella pneumoniae isolated from patients admitted in a tertiary care centre. MATERIALS AND METHODS: A total of 111 clinically significant Enterobacteriaceae isolates, resistant to cephalosporin subclass III were screened for carbapenemase production by the modified Hodge test. Minimum inhibitory concentration to imipenem and meropenem was determined and interpreted according to Clinical Laboratory Standards Institute 2011 criteria. Presence of bla NDM-1 was detected by polymerase chain reaction. To ascertain clonal relatedness, random amplification of polymorphic deoxyribonucleic acid (RAPD) was carried out for representative NDM-1 producers. RESULTS: bla NDM-1 was detected in 64 study isolates, of which 27 were susceptible to carbapenems. RAPD revealed a high degree of clonal diversity among NDM-1 producers except for a small clustering of isolates in the neonatal intensive care unit. CONCLUSION: There is extensive clonal diversity among the NDM-1 producing E. coli and K. pneumoniae. Hence, antibiotic selection pressure rather than horizontal transfer is probably an important operating factor for the emergence of NDM-1. This calls for increased vigilance, continuous surveillance and strict enforcement of antibiotic policy with restricted use of inducer drugs.

Endophthalmitis caused by Citrobacter species: a case series.

OBJECTIVE: To report the clinical profile and drug sensitivity patterns in Citrobacter endophthalmitis. DESIGN: Retrospective interventional case series. METHODS: Retrospective analysis of all the cases of Citrobacter endophthalmitis presenting to a tertiary referral ophthalmic centre in eastern India from January 2009 to December 2011 was done. RESULTS: Five cases were included in the study. Of these 5, 1 was posttraumatic, 1 was of endogenous origin, and the rest were postoperative. Vitreous surgery was done for 3 cases, whereas 1 patient declined surgery. The endogenous case received only intravitreal antibiotics. All isolates were sensitive to ciprofloxacin. Only 1 of the 5 isolates was sensitive to ceftazidime. Only 1 eye achieved a final visual acuity of 20/60 and the rest became phthisical at final follow-up. CONCLUSIONS: Citrobacter is a rare cause of endophthalmitis with poor outcome. Ciprofloxacin should be considered as the first line of treatment.

Endophthalmitis caused by Acinetobacter baumanni: a case series.

PURPOSE: To profile the etiology, clinical outcomes and drug sensitivity patterns in endophthalmitis caused by Acinetobacter baumanni. METHODS: Retrospective analysis of all the cases of Acinetobacter baumanni endophthalmitis presenting to tertiary referral care ophthalmic hospital in Eastern India from January 2009 to December 2011 were done. RESULTS: A total of four cases were included in the study. Out of the four cases one was post traumatic and the rest were post cataract surgery. All the cases underwent vitreoretinal surgical intervention followed by intravitreal antibiotics. A. Baumanni was isolated from vitreous in all the cases. Among all the drugs tested bacteria were found sensitive to ciprofloxacin (100 %) whereas all tested resistant to ceftazidime. Out of the four cases one had to be eviscerated, another developed retinal detachment post vitrectomy, one was phthisical at final followup, and only one patient achieved a vision of 20/200 with clear media and attached retina at final visit. CONCLUSION: A. Baumanni is a very rare cause of endophthalmitis with poor visual and anatomical outcomes. Ciprofloxacin should be considered as first the line intravitreal antibiotic.

Isolation of a variant human adenovirus identified based on phylogenetic analysis during an outbreak of acute keratoconjunctivitis in Chennai.

BACKGROUND & OBJECTIVES: Though several viruses are responsible for conjunctivitis, but human adenovirus (HAdV) is by far the most common cause. Epidemic conjunctivitis causes morbidity and early detection of aetiological agent is essential in preventing spread of disease as some of serotypes of adenoviruses cause a severe form of conjunctivitis. This study was undertaken to identify the causative agent of conjunctivitis outbreak in Chennai in 2010. METHODS: Conjunctival samples collected from 17 patients with conjunctivitis were subjected to virological investigations. Culture and PCR for detection of adenovirus and enterovirus were carried out. PCR positive products were further subjected for DNA sequencing. The nucleotide sequences of the hexons of isolates were analyzed by comparison with all 51 human adenovirus strains. Phylogenetic tree was constructed using DAMBE software. RESULTS: Among 17 patients, seven were positive for adenovirus by PCR on the direct specimen, none was positive for enterovirus. Eleven of 30 conjunctival swabs showed cytopathic effect in HEp-2 cell line and were confirmed as HAdV by PCR. The DNA sequence data of the 11 isolates had equal percentage of homology with HAdV 6 and 2 on blast analysis. On phylogenetic analysis with GeneBank data of 51 adenovirus strains, 11 isolates from patients during the outbreak of conjunctivitis formed a separate clade indicating a new variant strain. INTERPRETATION & CONCLUSIONS: Based on phylogenetic analysis it was concluded that the recent conjunctivitis outbreak that occurred in Chennai was caused by a variant adenovirus strain.

Complication of salmonella bacteremia in a case of treated fungal endophthalmitis.

This is to report a case of bacteremia caused by Salmonella typhi in a treated unilateral fungal endogenous endophthalmitis in an 18-year-old male from one of the South Asian countries. Microbiological and molecular investigations were carried out on the eviscerated material and routine blood culture was carried out. Direct examination of eviscerated material revealed the presence of fungal filaments. However, Salmonella typhi was isolated from both specimens, which was confirmed by Polymerase chain reaction targeting the 16SrRNA gene, sequencing, and random amplification of polymorphic DNA showed that they belonged to the same clone. The presence of Salmonella bacteremia in a treated unilateral fungal endophthalmitis, among young adult patients is rare and systemic symptoms should be investigated.

Determination of human cytomegalovirus pp65 antigenemia among renal transplant patients.

Human cytomegalovirus (HCMV) is an important cause of morbidity and mortality in immunosuppressed transplant recipients. Isolation of HCMV from peripheral blood leukocytes (PBLs) is considered a reliable marker of disseminated HCMV infection. HCMV pp65 antigenemia is widely used for monitoring CMV infection and guiding preemptive therapy. The aim of this study was to compare pp65 antigenemia with culture technique for detection of HCMV in PBLs among kidney transplant patients and also to determine the threshold value of significant pp65 antigenemiat. Fifty-one peripheral blood samples from post-renal transplant patients collected during August 2009 to March 2011 were processed for pp65 antigenemia assay. These were also tested for isolation of the virus by inoculation into human corneal fibroblast cells. The results of pp65 antigenemia and culture were compared to determine the clinical significance of pp65 antigenemia. HCMV was isolated in 21 cases. On comparing the pp65 antigenemia results with that of the viral isolation, a mean of 23 cells was determined to yield a positive isolation of HCMV. The values of pp65 antigenemia and isolation results were correlated (paired t-test, P = 0.0029). A pp65 count of 23 and above was considered significant in our clinical settings since we found that these clinical specimens yield positive culture result.

Chikungunya virus iridocyclitis in Fuchs' heterochromic iridocyclitis.

We are reporting a case of bilateral Fuchs' heterochromic iridocyclitis with chikungunya virus infection in the left eye. A 20-year-old female was presented with a past history of fever suggestive of chikungunya with bilateral Fuchs' heterochromic iridocyclitis and complicated cataract. She had a tripod dendritic pattern of keratic precipitates by confocal microscopy in the left eye with a stippled pattern of keratic precipitates in both eyes. The real-time polymerase chain reaction (RT-PCR) assay in the aqueous humor detected 98 copies/ml of chikungunya virus RNA. The patient underwent clear corneal phacoemulsification with in-the-bag intraocular lens implantation in the left eye with a good visual outcome. This is the first report where the presence of chikungunya virus RNA has been associated with a case of bilateral Fuchs' heterochromic iridocyclitis.

Application of PCR based-RFLP for species identification of ocular isolates of methicillin resistant staphylococci (MRS).

BACKGROUND & OBJECTIVE: Early detection of methicillin resistant staphylococci (MRS) from clinical specimens enables institution of appropriate antimicrobial therapy. Limited information is available on speciation of MRS. This study was undertaken to compare results of conventional and molecular methods in detection of methicillin resistance (MR) and application of PCR-restriction fragment length polymorphism (RFLP) and DNA sequencing for speciation of ocular isolates of MRS. METHODS: A total of 110 consecutive ocular staphylococcal isolates were screened for MR. MRS was speciated by PCR-RFLP of gap gene and results were confirmed by DNA sequencing. All isolates were processed within 48 h of isolation. A single colony of bacterium, stocked as stab cultures in Hyer's and Johnson agar, was stored at 4 degrees C and sub-cultured at every 15 days interval. RESULTS: Seventy (63.6%) of 110 isolates were identified as MRS and 40 (36.4%) were MSS by conventional and molecular method (100% correlation). Of the 70 MRS, 18 (25.7%) were Staphylococcus aureus, remaining 52 (74.3%) were CNS by conventional and molecular method (100% correlation). PCR-RFLP of gap gene identified 18 (25.71%) MRS as S. aureus, 11 (15.71%) S. epidermidis, 27 (38.57%) S. haemolyticus, 6 (8.57%) S. cohnii subsp. urealyticum, 6 (8.57%) S. equorum, 1 (1.42%) S. xylosus and 1 (1.42%) S. hominis. INTERPRETATION & CONCLUSION: Overall rate of isolation MRS was 63.6 per cent and were predominantly isolated from conjunctival swab (23.6%) and donor corneal scleral rim (23.6%) of non hospitalized patients indicating their community origin. Detection of MR by mecA gene was easier and less time consuming compared to conventional methods. Speciation of MRS was possible by gap gene PCR - RFLP and the predominant MRS in our study was S. haemolyticus.

Relative efficiency of polymerase chain reaction and enzyme-linked immunosorbant assay in determination of viral etiology in congenital cataract in infants.

BACKGROUND: Perinatal viral infections of fetus are among the leading causes of congenital cataract and identifying the viral etiology is important. OBJECTIVES: To detect the presence of Rubella virus (RV), herpes simplex virus (HSV) and cytomegalovirus (CMV) in lens aspirate specimens obtained from patients with congenital cataract and relate the results with serology. SETTING AND DESIGN: Prospective study carried out in tertiary care hospital. MATERIALS AND METHODS: Fifty lens aspirates from 50 infants with congenital cataract were subjected to HSV, RV isolation and polymerase chain reaction (PCR) for detection of HSV and CMV. Reverse transcription polymerase chain reaction (RT-PCR) was applied for RV detection. Peripheral blood specimens were screened for anti-HSV, RV and CMV antibodies by enzyme-linked immunosorbant assay (ELISA). RESULTS: Rubella virus was detected in nine (18%) lens aspirates, by nRT-PCR which includes six positive by culture. HSV-2 DNA was detected in nine other lens aspirates, while CMV was not detected by PCR. Serological results did not correlate with the presence of viruses in the lens aspirates. This is the first report of detection of HSV-2 DNA in cases of congenital cataract. CONCLUSIONS: Cytomegalovirus may not be playing a significant role in causation of congenital cataract. The role of serology in identifying causative viral infection for congenital cataract needs to be re-evaluated.

Population dose from indoor gamma exposure in the dwellings around Kudankulam nuclear power plant.

To assess the population dose due to the natural background radiation around the upcoming Kudankulam nuclear power plant, a systematic investigation has been carried out by measuring the indoor gamma dose. In total, 159 dwellings have been selected around the Kudankulam nuclear power plant area i.e. in Radhapuram and Nanguneri taluk (sub-districts) for the measurement. The geometric mean value of indoor gamma dose rate is 305 +/- 48 nGy h(-1) and 273 +/- 50 nGy h(-1) in Radhapuram and Nanguneri taluks (sub-districts), respectively. The annual effective dose due to indoor gamma radiation to the population has been found to be 1.5 mSv and 1.36 mSv in Radhapuram and Nanguneri taluks, respectively.

Gamma dose measurement in dwellings of Agastheeswaram Taluk of Kanyakumari district, lying 30 km radius from Kudankulam nuclear power plant site.

Most of the building materials contain naturally occurring radioactive elements, the most important of which are potassium (40)K and the members of two natural radioactive series, which can be represented by the isotopes of thorium (232)Th and Uranium (238)U. The presence of these radioisotopes in the materials causes external exposure to the people who live in the building. In addition, the disintegration of Uranium (238)U increases the concentration of radon gas (222)Rn and of its daughters in the house. So a systematic indoor gamma dose measurement has been performed in the dwellings of Agastheeswaram Taluk of Kanyakumari district, which is lying within the 30 km radius from the upcoming Kudankulam nuclear power plant site. The geometric mean of annual absorbed dose from gamma radiation in dwellings has been found to be 278 nGyh(-1). The seasonal variation of indoor gamma dose measurements has also been studied. Significant differences have been observed in dwellings built of different materials such as concrete, tiled, etc.

Studies on the distribution of 210Po and 210Pb in the ecosystem of Point Calimere Coast (Palk Strait), India.

A systematic study on the natural radionuclides such as 210Po and 210Pb in the environmental matrices of Point Calimere ecosystem has been undertaken to establish a baseline data on the radiation profile of Point Calimere environment. The environmental samples such as water, sediment and biota (seaweeds, crustaceans, molluscs and fish) have been subjected to analyses. It has been observed that the concentration of 210Po and 210Pb in the water samples of Point Calimere to be 0.5 mBq/l and 1.3 mBq/l, respectively. The soft tissues of the organisms accumulated higher 210Po content while shells and bones contained more 210Pb. The bivalve molluscs Meretrix casta have been identified to accumulate higher concentration of 210Po suggesting that they could serve as bio-indicator of radionuclides like 210Po in the Point Calimere ecosystem. The concentration factor of 210Po for the biotic components ranged from approximately 10(3) to 10(6) while for 210Pb it ranged from approximately 10(3) to 10(5).

Aciclovir resistance among Indian strains of Herpes simplex virus as determined using a dye uptake assay.

Resistance to aciclovir (ACV) among Herpes simplex virus (HSV) isolates is increasingly being reported in the literature particularly in immunocompromised patients. However, there is only limited data available from India despite widespread use of ACV in our hospital. A cross-sectional study was hence conducted to determine the aciclovir (ACV) susceptibility of HSV 1 and 2 isolates using a dye uptake (DU) assay. This study showed a 3.0% prevalence of ACV resistance among HSV-1 strains (2/66, median IC 50 0.098 microg/mL) while in HSV-2 strains, it was 7.8% (5/64, median IC 50 0.195 microg/mL). The IC 50 for the HSV-1 and HSV-2 strains resistant to ACV was greater than or equal to 6.25 microg/mL.