Efficacy and Toxicity Profile of Methotrexate Chloroquine Combination in Treatment of Active Rheumatoid Arthritis.

BACKGROUND: The present study was conducted to study the efficacy and toxicity profile of methotrexate chloroquine combination in treatment of active rheumatoid arthritis. METHODS: 24 patients of rheumatoid arthritis confirming to revised American Rheumatism Association (ARA) criteria were studied prospectively for twenty months. Clinical evaluation was made every 3 months. Clinical disease variables measured at each visit were number of joints with swelling, number of joints with tenderness and pain, duration of morning stiffness and physician and patient assessment of disease activity. Blood counts, liver function tests and other adverse effects due to drugs were monitored every 2 months. RESULTS: 10 patients demonstrated more than 50% improvement. 4 patients withdrew from study, 2 because of excessive nausea and vomiting and 2 because of noncompliance. Other side effects noted were hyperpigmentation, photosensitivity, skin rashes, raised transaminases and stomatitis. CONCLUSION: Methotrexate chloroquine combination has good efficacy and toxicity profile. Gastrointestinal side effects are most common and usually responsible for the discontinuation of the drugs.

Epididymal histoplasmosis diagnosed by isolation of Histoplasma capsulatum from semen.

An autochthonous case of epididymal histoplasmosis masquerading as tuberculosis in a 55-year-old male patient is reported from India. It was diagnosed by culture of Histoplasma capsulatum from semen and by demonstration of the fungus upon re-examination of epididymal biopsy sections previously misinterpreted as tuberculous granuloma. The patient's main complaints were painful epididymal swelling, occasional fever and cough. He was treated successfully by excision of epididymis and vas deferens combined with amphotericin B therapy. This is believed to be the first case of epididymal histoplasmosis to be reported outside the American continent and the fourth of its type reported in the English literature. The case is also noteworthy in that H. capsulatum was isolated for the first time from semen, and it underlines the importance of mycological culture of semen specimens for diagnosis of genitourinary infections of obscure etiology.

Coordination of germ tube formation and surface antigen expression in Candida albicans.

If the determinants of shape and cell wall topography are independently regulated and induced in germ tube formation in Candida albicans, these processes may be separable in a non-germ tube forming strain. The expression of several preferentially expressed hyphal surface components in a parental, non-germ tube forming variant, and a germ tube forming revertant strain were examined by indirect immunofluorescence. The proportion of germ tubes expressing the determinants and the morphological localization of expression was similar. Few yeast cells in germ tube cultures bound probes and there was no increase in binding by yeast cells of the variant strain. Extraction with beta-mercaptoethanol prior to analysis had little effect on probe binding and the shape of yeast cells were similar. These observations suggest the ability to promote apical expansion in germ tube formation and surface expression of certain markers were coordinately regulated.

Efficacy of seed-based media for the mould-yeast conversion of Blastomyces dermatitidis.

The efficacy of 20 seed-based media is reported for the in vitro mould-yeast conversion of Blastomyces dermatitidis, employing pharmamedia agar, peptone glucose agar, glucose agar and water agar as controls. The mould-yeast conversion varied significantly according to the culture medium, fungal strains and incubation period (p less than 0.01). Garden-pea, chick-pea, cow-pea, soybean, peanut, green gram, French bean, lentil, okra and cottonseed converted all of the 7 B. dermatitidis test strains after 5 days of incubation at 37 degrees C. Although the efficacy of many of these seed media was found to be at par with pharmamedia agar - a commercial cottonseed embryo-derived protein, garden-pea seed agar is adopted because of the wider availability and low fat content of this seed. The recommended composition of the medium comprises 2% aqueous seed extract, 2% glucose and pH 6-7. Only nigerseed and sunflower seeds failed to support the conversion of B. dermatitidis. Of the control media, peptone glucose agar, glucose agar and water agar did not support the conversion of 2 of the B. dermatitidis test strains. The mechanism underlying variable mould-yeast conversion of B. dermatitidis on seed-based media is not clearly understood. However, most of the seeds supporting excellent mould-yeast conversion are known for their high protein content. The conversion was apparently not affected by the fat content of the seeds or by incorporation of glucose in the medium.

Efficacy of brain heart infusion-egg albumen agar, yeast extract phosphate agar and peptone glucose agar media for isolation of Blastomyces dermatitidis from sputum.

The efficacy of brain heart infusion (BHI)-egg albumen agar, yeast extract phosphate agar and several modified peptone glucose agar media was evaluated for isolation of Blastomyces dermatitidis from sputum concomitantly seeded with the yeast form of the pathogen and Candida albicans. Based upon high per cent culture positivity of sputum, improved recovery (CFU/ml) of the seeded inoculum, faster growth rate of B. dermatitidis and low level of contamination, BHI-egg albumen agar, followed by yeast extract phosphate agar are recommended as the media of choice for the isolation of B. dermatitidis from contaminated clinical specimens.

Cottonseed extract versus pharmamedia for the in vitro mould-yeast conversion of Blastomyces dermatitidis.

A cottonseed medium, based on a 1% aqueous extract of seeds of any of the eight indigenously available varieties of cotton belonging to Gossypium hirsutum or Gossypium arboreum, was evaluated for the mould-yeast conversion of Blastomyces dermatitidis in vitro, and compared with pharmamedia agar as a control. The cottonseed agar was found to be as efficient as pharmamedia agar for the mould-yeast conversion of the 19 B. dermatitidis strains tested. Therefore, cottonseeds provide an adequate and inexpensive substitute for pharmamedia for the mould-yeast conversion of B. dermatitidis. It should be noted that the term 'cottonseed' medium has been used rather loosely in the literature and that this medium is based on pharmamedia and not on a cottonseed extract. The authors suggest that media based on pharmamedia should be referred to as pharmamedia agar and the use of the term 'cottonseed' should be restricted to media containing cottonseed extract.

Isolation and partial characterization of a plasma membrane-enriched fraction of Blastomyces dermatitidis.

The isolation and characterization of a plasma membrane-enriched fraction is reported from the pathogenic fungus Blastomyces dermatitidis employing a simplified and inexpensive procedure. The yeast cells, suspended in an osmotic stabilizer buffer, were subjected to gentle mechanical disruption followed by a three-step differential centrifugation to obtain a 40,000 x g sediment. This fraction showed 2.7- and 3.3-fold relative enrichment of the plasma membrane marker enzymes Mg2(+)-ATPase and 5'-nucleotidase, respectively, with modest cross-contamination of other intracellular membranes. The fraction comprised approximately 49% protein, 31% carbohydrate, 11% phospholipids and 9% sterol with sterol to phospholipid molar ratio of 1.1. Transmission electron microscopy of the plasma membrane-enriched fraction showed homogeneous vesicles with a tripartite unit membrane of 115 A thickness.

Efficacy of immunoprecipitation methods for rapid serodiagnosis of allergic bronchopulmonary aspergillosis.

The relative efficacy of Ouchterlony's double diffusion (ODD), Wadsworth's gel diffusion (WGD) and counter-immunoelectrophoresis (CIE) is reported for serodiagnosis of allergic bronchopulmonary aspergillosis (ABPA). Of the 50 sera taken from patients with confirmed or suspected ABPA, serum precipitins were demonstrated against Aspergillus fumigatus by CIE, WGD and ODD in sera from 34, 31 and 25 patients, respectively. The results were negative by the three methods in sera taken from 10 healthy volunteers. It is concluded that CIE followed by WGD is more efficacious than ODD for serodiagnosis of ABPA.

In vitro interactions between Blastomyces dermatitidis and other zoopathogenic fungi.

The results of in vitro interactions between colonies of Blastomyces dermatitidis and six other zoopathogenic fungi are reported. The interactions were found to range from neutral with Histoplasma capsulatum and Candida albicans to strongly antagonistic with Microsporum gypseum, Pseudallescheria boydii, and Sporothrix schenckii, and including lysis by Cryptococcus neoformans. These observations suggest that interactions between zoopathogenic fungi may be one of the biotic factors likely to influence the occurrence of B. dermatitidis in natural systems.

Survival of Blastomyces dermatitidis in the gastrointestinal tract of an orally infected insectivorous bat, Rhinopoma hardwickei hardwickei Gray.

The survival of Blastomyces dermatitidis with markedly reduced population, in the gastrointestinal tract and faeces of Rhinopoma hardwickei orally infected with 2.5 X 10(5) colony forming units of the fungus, is reported. B. dermatitidis was cultured from the stomach, intestine and faeces up to 16-24 h and from rectum up to 48 h post infection. The results demonstrate that orally infected R. hardwickei hardwickei bats transiently shed B. dermatitidis through their faeces but the significance of this route of environmental dissemination requires further evaluation.

Blastomyces dermatitidis in bats: first report of its isolation from the liver of Rhinopoma hardwickei hardwickei Gray.

Blastomyces dermatitidis is reported for the first time from the liver of Rhinopoma hardwickei hardwickei Gray (the 'lesser rat-tailed bat'); it was cultured from one of 46 samples of the bat captured on December 10, 1982, from the basement of Safdar-Jang Tomb, a historical monument in New Delhi. The fungus was not found in 581 other bats representing R. hardwickei hardwickei, three more insectivorous and one frugivorous species investigated from several sites in Delhi and New Delhi metropolitan areas. The identity of the isolate was based upon its macroscopic and microscopic cultural morphology, dimorphic character and verification of pathogenicity for white mice. It was further confirmed by determining the capacity of the isolate to produce the 'A' exoantigen specific for B. dermatitidis. The infected bat did not manifest any obvious clinical signs and symptoms of illness. Its visceral organs were free from macroscopic lesions, and histopathologically none of them including the liver, revealed any fungal elements or tissue response. B. dermatitidis was not found in any of the 34 samples of bat guano investigated by direct culture or mouse-inoculation technique. The results reinforce the available evidence for the endemic occurrence of B. dermatitidis in India and focus on the possible role of R. hardwickei hardwickei as a natural host or vector for this pathogen.

Prevalence of Basidiobolus ranarum Eidam in the intestinal tract of an insectivorous bat, Rhinopoma hardwickei hardwickei Gray, in Delhi.

The prevalance of Basidiobolus ranarum Eidam is reported from the intestinal contents of 14 (7%) of 200 bats belonging to Rhinopoma hardwickei hardwickei Gray ('the lesser rat-tailed bat'), an insectivorous species captured from Delhi area. Eleven of the positive bats were captured during August/September whereas the remaining three came from collections made during November or April. No macroscopic or microscopic lesions were found in the intestine of the bats yielding B. ranarum. This is believed to be the first report on the association of B. ranarum with bats. A more extensive sampling of bat species including their guano is indicated to elucidate the role of Chiroptera in the biological cycle of B. ranarum.