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Background: Chronic nonhealing wounds are difficult to manage. Various substances are being used to heal these wounds. We sought to observe the effects of autologous epidermal cell suspension dressings on chronic nonhealing ulcers. Methods: We enrolled patients of the wound clinic at University Hospital, Varanasi, India, with nonhealing wounds of more than 6 weeks' duration. We treated the wound beds with sterile dressings and antibiotics until the swab cultures became sterile. We prepared autologous epidermal cell suspensions from skin grafts and used them on the ulcer beds along with Vaseline gauze dressings. Follow-up visits with patients occurred weekly for assessment of wound healing and other changes. Results: Fifteen patients enrolled in our study. Of these, 6 patients had completely healed at 12 weeks, 1 patient at 16 weeks and 2 patients at 20 weeks after treatment. We observed a slow healing response in 6 patients, of whom 1 patient had healed completely at 32 weeks and another at 48 weeks. One patient needed skin grafting, and 3 patients were lost to follow-up. Conclusion: Autologous noncultured epidermal cell suspension transplantation seems to be an effective, simple and time-saving method to treat chronic nonhealing wounds. Contexte : La prise en charge des plaies chroniques qui ne guerissent pas pose un defi. Diverses substances existent pour favoriser la cicatrisation de ces plaies. Nous avons voulu observer les effets de pansements a base de suspension de cellules epidermiques autologues sur des ulceres chroniques. Methodes : Pour cette etude, nous avons recrute les participants parmi les patients de la clinique de traitement des plaies de l'Hopital universitaire de Varanasi, en Inde; les patients choisis avaient des plaies qui persistaient depuis plus de 6 semaines. Nous avons traite les plaies au moyen de pansements steriles et d'antibiotiques jusqu'a ce que les cultures de specimens soient negatives. Nous avons prepare des suspensions de cellules epidermiques autologues a partir de greffons cutanes et les avons appliquees sur les ulceres avec des pansements de gaze impregnes de Vaseline. Nous avons ensuite procede a un suivi hebdomadaire des patients pour verifier la cicatrisation de leurs plaies et autres changements. Resultats : Nous avons inscrit 15 patients a notre etude. Parmi eux, 6 ont presente une cicatrisation complete a 12 semaines, 1, a 16 semaines, et 2, a 20 semaines suivant le traitement. Nous avons observe une cicatrisation lente chez 6 patients, dont un a connu une guerison complete a 32 semaines et un autre, a 48 semaines. Un patient a du subir une greffe de peau et 3 n'ont pu etre retraces au suivi. Conclusion : L'inoculation d'une suspension de cellules epidermiques autologues non cultivees semble une methode efficace, simple et rapide de traitement des plaies chroniques qui ne guerissent pas., Wound healing is a dynamic biologic process to restore normal anatomy and physiology of the integumentary system and includes overlapping phases of inflammation, proliferation and maturation. (1) A prolonged and [...]

Abstract Background Enteric protozoa and sporozoa have emerged as important opportunistic parasites and can cause fatal infections in AIDS patients. The line of treatment being different for them necessitates an accurate and prompt identification of these to avoid empirical treatment. In this study which is the first of its kind from India we did a comprehensive evaluation of different techniques, comparing them on the basis of the attributes like yield, cost, time taken, expertise and infrastructure. For the first time combination of Calcoflour White and DAPI, a nuclear stain, were used to identify Microsporidia spp. Thus, a diagnostic protocol was devised for rapid, sensitive and cost effective identification of the opportunistic enteric protozoa. Results The organisms isolated from the stool samples of the cases (450 HIV patients) were predominantly Cryptosporidium spp., Microsporidia spp. and Cyclospora spp. Interestingly, the control group (200 relatives of the patients who were HIV negative) showed a high incidence (21%) of Cryptosporidium spp. We found a significant increase in the sensitivity of microscopy in detecting Cryptosporidium spp. and Cyclospora spp. after formol ether concentration. Kinyoun's staining was better compared to Modified safranin staining for Cryptosporidium spp. identification. Although ELISA had a sensitivity of 93.25% and specificity of 97% for Cryptosporidium spp. detection, we ranked Kinyoun's staining better than ELISA because it is not affordable to most of our patients. For detecting Cyclospora cayetanensis, autoflourescence was the easiest and most cost effective method followed by Safranin technique. Combination of Calcoflour White stain and DAPI gave good results for the identification of Microsporidia spp. We assessed the above techniques and graded the attributes in the following descending order: cost effectiveness, sensitivity, ease of use and interpretation, time taken for the procedure and batch testing. Conclusion Thus, we conclude that a combination of minimum three procedures should be carried out for the screening of stool specimens of HIV positive patients. Kinyoun's staining should be made mandatory for every diarrheal stool sample from HIV patients. Also every laboratory should assign its own value to the attributes and apply Multiattribute utility theory or the Analytical hierarchy process to decide the most appropriate methodology.

Abstract Background Protozoan infections are the most serious among all the superimposed infections in HIV patients and claim a number of lives every year. The line of treatment being different for diverse parasites necessitates a definitive diagnosis of the etiological agents to avoid empirical treatment. Thus, the present study has been aimed to elucidate the associations between diarrhoea and CD4 counts and to study the effect of HAART along with management of diarrhoea in HIV positive patients. This study is the first of its kind in this area where an attempt was made to correlate seasonal variation and intestinal protozoan infestations. Methods The study period was from January 2006 to October 2007 wherein stool samples were collected from 366 HIV positive patients with diarrhea attending the ART centre, inpatient department and ICTC of S.S. hospital, I.M.S., B.H.U., Varanasi. Simultaneously, CD4 counts were recorded to assess the status of HIV infection vis-à-vis parasitic infection. The identification of pathogens was done on the basis of direct microscopy and different staining techniques. Results Of the 366 patients, 112 had acute and 254 had chronic diarrhea. The percentages of intestinal protozoa detected were 78.5% in acute and 50.7% in chronic cases respectively. Immune restoration was observed in 36.6% patients after treatment on the basis of clinical observation and CD4 counts. In 39.8% of HIV positive cases Cryptosporidium spp. was detected followed by Microsporidia spp. (26.7%). The highest incidence of intestinal infection was in the rainy season. However, infection with Cyclospora spp. was at its peak in the summer. Patients with chronic diarrhea had lower CD4 cell counts. The maximum parasitic isolation was in the patients whose CD4 cell counts were below 200 cells/μl. Conclusion There was an inverse relation between the CD4 counts and duration of diarrhea. Cryptosporidium spp. was isolated maximum among all the parasites in the HIV patients. The highest incidence of infection was seen in the rainy season.

A letter to the editor is presented in response to an article about medical study on superficial fungal infection tinea nigra.

The present study was designed to determine antibiotic resistance rates and patterns and its correlation with enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) whole genome analysis of Pseudomonas aeruginosa obtained from clinical and environmental sources. In order to determine the possibility of clonality in the bacteria and to track the resistance markers for a better understanding of the epidemiology of drug resistance. A total of 500 strains, 100 each from clinical, water, animal, plant and soil sources were subjected to antibiogram analysis by disc diffusion method. Seventy-five randomly selected strains, 15 each of the five sources were subjected to ERIC-PCR analysis. Clinical isolates were more resistant to combinations of very high numbers of drugs as compared to isolates from other sources. Weak clonality was observed in P. aeruginosa by ERIC-PCR method with 80% of the clinical strains belonging to only 3 clones. It could be concluded that it is the drug selection pressure in clinical environment that is causing the accumulation of drug resistance against many antimicrobials. Furthermore, P. aeruginosa does have clonal expansion. Further studies are warranted to confirm the results.

Salmonella enterica serotype Typhi (S. Typhi) is a causative organism of typhoid fever. A number of Salmonella serovars express a capsular polysaccharide antigen known as Vi, the biosynthetic and export proteins of which are encoded within the viaB locus of Salmonella Pathogenicity Island -7 (SPI-7). SPI-7 is inserted between two partially duplicated copies of tRNA -pheU gene. We have investigated the frequency of viaB operon deletion and loss of SPI-7 due to storage of strains collected during the period 1987-2006 by PCR amplification of fliC (for confirmation of serotype Typhi), tviB (for status of viaB operon) and tRNA -pheU (for absence of SPI-7). All 111 isolates were observed with positive amplification of 495 bp amplicon for fliC. A total of 36 isolates were negative for Vi by agglutination while 39 were negative for viaB operon. Interestingly, 106 isolates were found to have SPI-7. The 5 SPI-7 negative isolates were isolated during recent years. Long-term storage and repeated culture had little or no effect on SPI-7, as none of the 18 isolates recovered from blood before 1997 lacked SPI-7. On the other hand, loss of viaB operon was directly proportional to duration of storage. Thus, it is proposed that stability of Vi gene is dependent on the presence of selection pressure.

Indiscriminate use of antibiotics for infected chronic wounds is a global problem that may contribute to delayed healing and the development of drug-resistant micro-organisms. A prospective, descriptive cohort study of 50 male and female outpatients (mean age 52.50 [+/- 14.84] years, range 18-90) with 52 chronic wounds was conducted to investigate the microbiological profile and prevalence of drug-resistant strains in chronic nonhealing wounds to develop an evidence-based approach to antibiotic therapy until drug sensitivity reports are available. Mean wound duration was 8.23 (+/- 12.35) months (range 1.5-72), average wound size was 29.70 (+/- 37.83) cm, and most patients had a lower extremity wound and diabetes mellitus (n = 20). Pus and tissue samples were cultured and tested. Most (45) wounds contained a single organism and nine different genera were isolated. Of those, 39 were Gram-negative and 11 were Gram-positive (z = 5.50, P = <0.001). The most common organisms were Pseudomonas (21 wounds) and Escherichia coli (eight wounds). Pseudomonas aeruginosa was more common in patients with diabetes mellitus, in lower extremity ulcers, and in ulcers >20 cm2 (z-test, P <0.05). The presence of two organisms was more commonly observed in postsurgical/traumatic wounds. Ten (10) out of 55 pathogens (18.18%) isolated were drug-resistant, including Pseudomonas (seven), methicillin-resistant Staphylococcus aureus (one), and extended-spectrum beta lactamase (two--E. coli and Citrobacter). Most (70%) drug-resistant pathogens were obtained from persons with diabetes mellitus. Overall sensitivity to piperacillin and tazobactum combination was high. Because the prevalence of monomicrobial flora in chronic wounds is high, if a wound infection is suspected, empiric therapy should target the most prevalent flora. The high rate of drug-resistant Pseudomonas and MRSA strains should discourage antibiotic use in chronic ulcers before obtaining culture results.