Paediatric dacryocystorhinostomy

Of 258 cases of dacryocystorhinostomy performed on children in the period September 1981 to September 1991, 130 were for simple, unresolved congenital nasolacrimal duct obstruction. Other indications for surgery included punctal agenesis, lacrimal fistula, post-traumatic and post-inflammatory canalicular obstruction. Of 177 children without canalicular pathology, 171 (96%) were relieved of symptoms with one operation, without canalicular intubation. Of 81 cases with canalicular disease, 55 of 70 (79%) who underwent DCR plus canalicular intubation, and 10 of 11 who underwent DCR plus Lester-Jones tube, were substantially improved with one operation. No child required peroperative or postoperative blood transfusion. Dacryocystorhinostomy in childhood, in experienced surgical hands, is a safe procedure, achieving relief of symptoms in most cases, particularly in the absence of canalicular disease.     

Comparison of a modified adherence assay with existing assay methods for identification of enteroaggregative Escherichia coli.

Localized, diffuse, and aggregative adherence patterns of Escherichia coli identified with specific DNA probes were compared in cell culture adherence assays by using the Center for Vaccine Development, Baltimore, method, the University of Texas Medical School, Houston (UTH), method, and a modified UTH method. Increasing postwash incubation time from 2 to 4 h in the modified UTH method allowed identification of enteroaggregative E. coli, which was otherwise not identified by the original UTH method.     

Pathogenesis of Providencia alcalifaciens-induced diarrhea.

Providencia alcalifaciens is a member of the family Enterobacteriaceae. There are reports that P. alcalifaciens can cause diarrhea, but the mechanism(s) by which it causes diarrhea is known. We studied P. alcalifaciens isolated from a child and two adults with diarrhea for enteropathogenicity. The three isolates did not exhibit any characteristic adherence to cultured HEp-2 cell monolayers, and they did not produce enterotoxins, cytotoxins, or keratoconjunctivitis in the Sereny test. Two isolates invaded cultured HEp-2 cell monolayers, producing localized bacterial clusters and actin condensation. The pattern of actin condensation was different from that produced by enteropathogenic Escherichia coli but similar to that produced by Shigella flexneri. Invasion and actin condensation were poor for the third isolate. Histology of adult rabbit small intestinal loops inoculated with all three isolates revealed bacterial attachment to, penetration of, and microulcer formation on the surface epithelium and hyperemia, edema, and polymorphonuclear cell infiltration of lamina propria. All the isolates produced diarrhea in rabbits with removable intestinal ties, and some of these rabbits developed hindlimb paralysis. Intestinal histology of the rabbits with removable intestinal ties which developed diarrhea showed changes similar to that in adult rabbits on which ileal loop assays had been performed. Transmission electron microscopy of intestinal tissues also confirmed tissue penetration by the isolates. Nerve tissue histology of two rabbits that developed hindlimb paralysis showed focal mononuclear cell infiltration around peripheral nerve sheaths. It is concluded that some strains of P. alcalifaciens are enteropathogenic and that they cause diarrhea by invading the intestinal mucosal epithelium. However, the relevance to human disease of the hindlimb paralysis observed in this animal model is not clear.     

Antimicrobial susceptibilities and plasmid contents of Neisseria gonorrhoeae isolates from commercial sex workers in Dhaka, Bangladesh: emergence of high-level resistance to ciprofloxacin

Commercial sex workers (CSWs) serve as the most important reservoir of sexually transmitted diseases (STD), including gonorrhea. Periodic monitoring of the antimicrobial susceptibility profile of Neisseria gonorrhoeae in a high-risk population provides essential clues regarding the rapidly changing pattern of antimicrobial susceptibilities. A study concerning the prevalence of gonococcal infection among CSWs was conducted in Bangladesh. The isolates were examined with regards to their antimicrobial susceptibility to, and the MICs of, penicillin, tetracycline, ciprofloxacin, cefuroxime, ceftriaxone, and spectinomycin by disk diffusion and agar dilution methods. The total plasmid profile of the isolates was also analyzed. Of the 224 CSWs, 94 (42%) were culture positive for N. gonorrhoeae. There was a good correlation between the results of the disk diffusion and agar dilution methods. Some 66% of the isolates were resistant to penicillin, and 34% were moderately susceptible to penicillin. Among the resistant isolates, 23.4% were penicillinase-producing N. gonorrhoeae (PPNG). 60.6% of the isolates were resistant and 38.3% were moderately susceptible to tetracycline, 17.5% were tetracycline-resistant N. gonorrhoeae, 11.7% were resistant and 26.6% had reduced susceptibility to ciprofloxacin, 2.1% were resistant and 11.7% had reduced susceptibility to cefuroxime, and 1% were resistant to ceftriaxone. All PPNG isolates contained a 3.2-MDa African type of plasmid, and a 24.2-MDa conjugative plasmid was present in 34.1% of the isolates. Since quinolones such as ciprofloxacin are recommended as the first line of therapy for gonorrhea, the emergence of significant resistance to ciprofloxacin will limit the usefulness of this drug for treatment of gonorrhea in Bangladesh.     

Fine-needle aspiration biopsy in diagnosis of follicular lymphoma: cytomorphologic and immunohistochemical analysis

Follicular lymphoma is a relatively uncommon type of non-Hodgkin's lymphoma (NHL) thought to derive from follicular center cells. There are limitations to fine-needle aspiration in the diagnosis of follicular NHL, and very few studies are available on it. We describe here the cytomorphologic features of eight biopsy-proven cases of follicular NHLs and five biopsy-proven cases of intermediate-grade NHLs. Five cases of reactive lymphoid hyperplasia (RLH) diagnosed on cytology were also studied cytomorphologically. Image morphometry was done in follicular and intermediate-grade NHLs, immunocytochemistry was done in follicular and intermediate-grade NHLs, and immunocytochemistry for bcl-2 expression was studied in five cases of follicular NHLs. The aspirate smears of follicular NHL showed the presence of higher grades, many to abundant (++ to +++) monomorphic lymphoid aggregates, with an increased number of mast cells. Lymphohistiocytic tangles and tingeable body macrophages were not observed in any of the cases. Cytomorphologically, all the cases of intermediate-grade NHL showed a predominantly monomorphic population of lymphoid cells with absent or few grades (+) of lymphoid aggregates. Image morphometric data analysis of each cell parameter for follicular and intermediate-grade NHLs were found to be statistically insignificant (P > 0.05) and hence of little help in diagnosis of follicular vs. low-grade NHLs. Immunocytochemistry for bcl-2 expression in follicular NHL showed variable results, ranging from 0-90% (expressed as bcl-2 index). Certain characteristic cytomorphological features such as the presence of monomorphic lymphoid cell aggregates and an increased number of mast cells are soft indicators for the diagnosis of follicular NHL on aspiration smears. Immunocytochemistry for bcl-2 expression may be of help in cases with clinical and cytological suspicion of lymphoma. Image morphometry has a small role in this regard.     

Covalent immobilization of proteins onto photoactivated polystyrene microtiter plates for enzyme-linked immunosorbent assay procedures

Enhancement of the speed and sensitivity of an ELISA technique was achieved by doing it on a polystyrene microtiter plate preactivated by a simple photochemical reaction. Immobilization of Epicoccum nigrum antigen (allergenic antigen) or goat anti-rabbit IgG onto the photoactivated plates was found to occur in only 45 min with higher binding than that obtained through adsorption during the same period onto the untreated surface. Nearly 1.5-2-folds higher readings were obtained when the ELISA was carried out with the solid phase prepared on the photoactivated surface rather than on the untreated surface. Moreover, solid phases prepared on the activated surface could detect IgE (E. nigrum antibody) even at 1/50 (v/v) dilutions, whereas a solid phase prepared on the untreated surface failed to do so. Around three times higher ELISA values were obtained in the activated plate than the untreated plate when IgE was diluted to 1/5 (v/v). Such photoactivated surface could be of great importance in diagnostic tests involving the ELISA technique particularly to confirm false negative cases and for other immunoassays such as radioimmunoassay procedures.     

Comparison of mismatch amplification mutation assay with DNA sequencing for characterization of fluoroquinolone resistance in Neisseria gonorrhoeae

A mismatch amplification mutation assay (MAMA) was developed for identification of point mutations in quinolone resistance-determining region (QRDR) of gyrA at codons 91 and 95. MAMA PCR was used to detect mutations at codons 91 and 95 of gyrA in 117 Neisseria gonorrhoeae isolates (with ciprofloxacin MICs of 0.004 to >32 microg/ml) from Bangladesh during 1997 to 2001. The QRDR regions of the gyrA genes from 31 randomly selected isolates were sequenced, and the results were compared with those of MAMA PCR. Using mismatch PCR, a mutation at Ser91 could be detected in all 27 (resistant and intermediate) isolates, and an Asp95-to-Gly95 mutation could be detected in all 15 isolates, as detected by sequencing. MAMA PCR offers a simple, inexpensive, rapid, and easier alternative for detection of point mutations in fluoroquinolone resistance in N. gonorrhoeae.