Fluorescence of Candida in diagnosis of oral candidiasis.

Background: Many pathogenic fungi fluoresce in hematoxylin and eosin stained sections, and Papanicolaou (PAP)‑stained smears under ultraviolet illumination. In theory, this phenomenon could aid in the diagnosis of common fungal infections without the delay which is usually associated with special stains. Objective: To evaluate the role of fluorescence as a rapid screening technique for oral infections caused by Candida organisms in exfoliative smears of oral candidiasis. Materials and Methods: Two smears and one swab were collected from each of 62 clinically diagnosed cases of oral candidiasis. Smears were stained with (PAP) and periodic acid–Schiff stain (PAS). Both smears were evaluated under light microscopy (LM). Later, PAP smears were observed under fluorescent microscopy (PAP‑FM). The swab was inoculated on Sabouraud’s agar plate. Each technique was evaluated for sensitivity and specificity. Results: It was found that the PAS‑stained smears were more reliable for detection of Candida species than other methods (sensitivity = 100%; specificity = 66.7%). The PAP‑LM and PAP‑FM showed less sensitivity (67.9% and 85.7%) and specificity (66.7% and 33.3%), respectively. Combined results of both light and fluorescent microscopy of PAP (LM + FM) showed increased sensitivity (89.3%) but reduced specificity (16.7%). Conclusion: PAP autofluorescence is less sensitive than PAS, still it accentuates the distinct morphological features of Candida.

Isolation and purifecation of riboflavin binding protein from eagle egg-yolk (aquila hastate).

Riboflavin binding protein was purified from the egg yolk of Aquila hastate (Eagle). The protein was purified using DEAE Sepharose ion exchange chromatography followed by gel filtration on Sephadex G-100. The purity of the protein was judged by cylindrical and slab SDSPAGE techniques. Comparison of the mobility of RfBP with that of the standard molecular weight marker protein revealed that the RfBP had a molecular weight close to 29 Kd. Interestingly the RfBPs from hen egg yolk and eagle egg yolk had the same molecular weights as revealed by the SDS PAGE.

Isolation and evaluation of antifertility activity of total alkaloids from leaves of aegle marmelos in male albino rats (rattus norvegicus).

Aegle marmelos (Rutaceae) is being used in traditional medicine treatments, such as for intermittent fever, intestinal ailments, fertility control etc. It has been proved to be effective against several major diseases including cancer, diabetis and cardiovascular diseases. Although the plant is a well known male antifertility plant, till today only the crude extracts of the plant were screened for antifertility activity in male rats. The plant is rich in alkaloid content and Aegelenine, Marmeline and Skimmianine, are some of the alkaloids isolated so far, showed variety of pharmacological activities. In view of these facts, in the present study, total alkaloids have been isolated from leaves of A. marmelos and their effect on fertility of adult male albino rats (Rattus norvegicus) was investigated. Three different doses 20, 40, 80 mg/kg body weight of total alkaloids were orally administered to mature male albino rats (Wistar strain) of proven fertility (235-2450gr) for 60 days. On day 61, all the animals were sacrificed and the fertility and safety parameters were studied. Weights of all the major reproductive organs, accessory glands and sperm counts were significantly decreased in dose dependent manner suggesting the antifertility activity and serological parameters showed no significant changes in treated animals at the tested dose levels indicating the safety of long-term use of total alkaloidal fraction of A. marmelos.

Purification and characterization of riboflavin binding protein (RfBp) from hen (Gallus gallus) eggs using deae-sepharose column chromatography.

Riboflavin binding protein (RfBP) was isolated, purified and characterized from Hen (Gallus gallus) egg white and yolk using Sepharose column chromatography. The Rfbp was purified using DEAE-Sepharose ion exchange chromatography followed by gel filtration on Sephadex G-100. The protein content was estimated with Lowry method. The purity of the proteins was judged by SDS-PAGE technique. The protein migrated as a single band on SDS gel with a molecular weight of 29 kilodaltons. This is the first report on purification of this protein using DEAE-Sepharose column chromatography.

Comparison of potassium hydroxide mount and mycological culture with histopathologic examination using periodic acid-Schiff staining of the nail clippings in the diagnosis of onychomycosis.

BACKGROUND: Onychomycosis is a common problem noticed in clinical practice. Currently available standard laboratory methods show inconsistent sensitivity; hence there is a need for newer methods of detection. AIMS: This study involves comparison of standard laboratory tests in the diagnosis of onychomycosis, namely, potassium hydroxide mount (KOH mount) and mycological culture, with histopathologic examination using periodic acid-Schiff (PAS) staining of the nail clippings. METHODS: A total of 101 patients with clinically suspected onychomycosis were selected. Nail scrapings and clippings were subjected to KOH mount for direct microscopic examination, culture using Sabouraud's dextrose agar (with and without antibiotics) and histopathologic examination with PAS staining (HP/PAS). Statistical analysis was done by McNemar's test. RESULTS: Direct microscopy with KOH mount, mycological culture, and HP/PAS showed positive results in 54 (53%), 35 (35%), and 76 (75%) patients respectively. Laboratory evidence of fungal infection was obtained in 84 samples by at least one of these three methods. Using this as the denominator, HP/PAS had a sensitivity of 90%, which was significantly higher compared to that of KOH mount (64%) or mycological culture (42%). CONCLUSIONS: Histopathologic diagnosis with PAS staining of nail clippings was the most sensitive among the tests. It was easy to perform, rapid, and gave significantly higher rates of detection of onychomycosis compared to the standard methods, namely KOH mount and mycological culture.

Clinicoepidemiological profile and predictors of severe illness in young infants (< 60 days) reporting to a hospital in North India.

OBJECTIVES: To describe the clinical and epidemiological profile of young infants reporting to a hospital and assess previously proposed simple clinical signs for their value in enabling health workers to detect young infants with severe illness warranting hospital admission. METHODS: Observational study of infants less than 2 months of age presenting consecutively to a large public hospital in South Delhi who were evaluated by a health worker (nurse), on a standardized list of signs and symptoms, and the ability of these were evaluated against the need for hospital admission which was assessed by an independent pediatrician. RESULTS: Of the 1624 young infants triaged, 878 were enrolled into the study. Of these 100 (11%) were below 7 days of age, for whom the common reasons for seeking care were jaundice (52%), not feeding well (6%) and fever (5%). The remaining 778 (89%) were 7-59 days of age with respiratory symptoms as the main presenting complaints (29.1%). The primary clinical diagnoses in infants with serious illness needing admission to hospital in the age group <7 days (n = 66) were hyperbilirubinemia (56%) and sepsis (21%). In those between 7-27 days of age (n = 60), primary diagnoses were sepsis (27%), pneumonia (13%), diarrhea, dysentery or dehydration (10%), while in the age group 28-59 days of age (n = 47) pneumonia (40%), sepsis (19%) and diarrhea or dehydration (13%) were the common primary diagnoses. Signs that had at least a prevalence of 5% and were strong predictors for all the age categories studied were history of difficult feeding (OR 6.8 for 0-6 days, 15.1 for 2-27 days and 6.2 for 28-59 days age groups), not feeding well on observation (OR 13.7, 27.6 and 20.9 respectively for the 3 age groups), temperature > 37.5C (OR 21.8, 14.6 and 30.0 respectively for the 3 age groups) and respiratory rate > 60 per minute (OR 6.8, 15.1 and 21.0 respectively for the 3 age groups). Additional strong predictors with > 5% prevalence were history of convulsions (OR 7.9, only in 0-6 day age group), lethargy (OR 26.1, only in 7-27 day age group), and history of diarrhea (OR 3.0 for 2-27 days and 2.2 for 28-59 days age groups). CONCLUSIONS: Simple clinical signs are useful in hands of health worker for identifying neonates with serious illness warranting hospital admission. These will be of use in the further development of clinical algorithms for the national integrated management of childhood illnesses.

Technological application of an extracellular cell lytic enzyme in xanthan gum clarification

Uma enzima extracelular celulolítica produzida por Pseudomonas sp. foi ativa sobre células de Xanthomonas campestris mortas pelo calor. A atividade lítica causou a digestão enzimática de goma xantana de X. campestris. A digestão foi eficiente tanto para xantana nativa altamante viscosa (2,0 per center w/v) como para xantana comercial Sigma (2,5 per center w/v). Observações por microscopia eletrônica de varredura demonstraram a ação celulolítica sobre células de X. campestris.

Safety and efficacy of simultaneous bilateral primary combined trabeculotomy-trabeculectomy for developmental glaucoma.

PURPOSE: To establish the safety and efficacy of simultaneous bilateral primary combined trabeculotomy-trabeculectomy for developmental glaucoma. METHODS: We studied 109 consecutive patients who underwent planned simultaneous bilateral primary combined trabeculotomy-trabeculectomy for developmental glaucoma by a single surgeon from January 1990 through December 1999. The main outcome measures were postoperative intraocular pressure (IOP), corneal clarity and diameter, visual acuity, bleb characteristics, time of surgical failure and complications. Postoperative complications including endophthalmitis and anaesthetic morbidity and mortality were also analysed. RESULTS: The series consisted of 218 primary combined trabeculotomy-trabeculectomy surgeries during 109 anaesthesias. The mean follow-up period was 16.33 +/- 16.22 months. The IOP reduced from 26.4 +/- 5.9 mmHg to 13.5 +/- 4.5 mmHg, with a mean percentage reduction of 46.2 +/- 23.7 (P < 0.0001). The success (IOP < 16 mmHg) probabilities were 90.9%, 88.0% and 69.3% at first, second and third year respectively (Kaplan-Meier analysis). The success probability of 69.3% obtained at third year was maintained till 6 years of follow-up. One hundred and sixty six (76.1%) eyes had significant corneal oedema. Postoperatively, the cornea cleared in 93 (57.8%) eyes. Clinically, well functioning blebs were present in 114 of 171 eyes (66.6%). Postoperatively, 18 (8.3%) eyes developed shallow anterior chamber and 6 (33.3%) of them required surgical reformation. There was no incidence of endophthalmitis or any other sight-threatening complication. Of the anesthetic complications, apnea occurred in 17 (15.6%) patients and all were successfully resuscitated. The most serious post-anaesthetic complication was cardio-pulmonary arrest that occurred 5 hours postoperatively following aspiration during feeding in one child; this child could not be resuscitated. Two children had delayed recovery (2 and 4 hours respectively). The child who had delayed recovery by 2 hours survived and has completed 3 years of follow-up while the other child expired 48 hours later. CONCLUSION: Simultaneous bilateral primary combined trabeculotomy-trabeculectomy is safe and effective for developmental glaucoma. It obviates the need for long second anaesthesia with its attendant risks. It offers several other benefits to the patients and families.

Isolation of sheep anterior pituitary messenger RNA and its translation in a heterologous cell-free system.

A preparation rich in the specific messenger RNA involved in the synthesis of prolactin from sheep anterior pituitary glands was obtained by employing both the immunochemical and affinity techniques. A dose-dependent and efficient stimulation of protein synthesis by the isolated total pituitary RNA as well as poly (A) rich RNA were achieved with the reticulocyte system. The synthesis of prolactin as one of the translational products of this cell-free system was established by specific immunoprecipitation followed by resolution on sodium dodecyl sulphate polyacrylamide gel electrophoresis.