Molecular typing of Mycobacterium spp. isolates from Yemeni tuberculosis patients.

The study was done to characterize at the species level Mycobacterium spp. isolates from Yemeni pulmonary tuberculosis patients. Early-morning sputum samples were collected from 170 patients referred to the National Tuberculosis Institute in Sana'a city with suspected pulmonary tuberculosis. Samples were processed with Ziehl-Neelsen stain and cultured in Ogawa and Lowenstein-Jensen media. The rpoB gene target sequence was amplified using mutagenesis forward and reverse primers followed by Hindlll enzyme digestion. Of the 120 isolates analysed, 118 (98.3%) were identified as M. tuberculosis complex and 2 (1.7%) were identified as mycobacteria other than M. tuberculosis. The results showed that those 2 isolates were multi-drug resistant and the DNA sequencing analysis showed that the alignment of nucleic acid of DNA in isolates of mycobacteria other than M. tuberculosis was different from that of M. tuberculosis complex.

Cryptosporidiosis in children in a north Jordanian paediatric hospital.

We investigated the rate of infection by Cryptosporidium parvum among children from birth to 12 years attending Princess Rahma Teaching Hospital in Irbid, Jordan and evaluated various diagnostic methods. We collected single stool specimens from 300 children; 7 specimens were from children undergoing chemotherapy treatment for cancer. Diagnostic methods used for detection of infection were direct wet mount preparation, flotation concentration, cold Kinyoun Ziehl-Neelsen stain and direct immunofluorescence. We detected C. parvum oocysts in 112 samples (37.3%) using direct immunofluorescence, which showed the highest sensitivity. Source of drinking water appeared to be an important risk factor for transmission of infection. A higher incidence of infection was recorded during January-May, the rainy season.

Cryptosporidiosis in children in a north Jordanian paediatric hospital

We investigated the rate of infection by Cryptosporidium parvum among children from birth to 12 years attending Princess Rahma Teaching Hospital in Irbid, Jordan and evaluated various diagnostic methods. We collected single stool specimens from 300 children; 7 specimens were from children undergoing chemotherapy treatment for cancer. Diagnostic methods used for detection of infection were direct wet mount preparation, flotation concentration, cold Kinyoun Ziehl-Neelsen stain and direct immunofluorescence. We detected C. parvum oocysts in 112 samples [37.3%] using direct immunofluorescence, which showed the highest sensitivity. Source of drinking water appeared to be an important risk factor for transmission of infection. A higher incidence of infection was recorded during January-May, the rainy season

Ultrasonographic imaging of mycetoma.

INTRODUCTION: The ultrasonographic appearance of mycetoma is described in this prospective study. METHODS: One hundred patients with foot swellings had sonographic evaluation of the swelling and surgical excision within 2 weeks of ultrasonography. The histopathological findings were compared with the preoperative images. Some of the excised swellings and grains were also imaged and compared with the in vivo findings. RESULTS: The mycetoma grains, their capsules and the accompanying inflammatory granulomas have characteristic ultrasonographic appearances. In eumycetoma lesions, the grains produce numerous, sharp hyper-reflective echoes and there are single or multiple thick-walled cavities with no acoustic enhancement. In actinomycetoma, the findings are similar but the hyper-reflective echoes are fine, closely aggregated and commonly settle at the bottom of the cavities. None of the non-mycetoma foot swellings (which included lipoma, ganglion, foreign body granuloma and others) studied had these features. CONCLUSION: Ultrasonography is simple, non-invasive, quick, reproducible and acceptable to patients. Mycetoma has characteristic ultrasonographic features. Furthermore, ultrasonography delineates the extent of mycetoma more accurately than clinical examination alone.

Blood supply and vasculature of mycetoma.

The blood supply to the mycetoma lesion and its vasculature were studied in patients with various types of mycetoma using histological, ultrastructural, angiographic and sonographic techniques. The mycetoma lesion proved to be well vascularized. However, certain vascular abnormalities were demonstrated. In histological sections, the small arteries and arterioles showed medial muscular hypertrophy in 83%, intimal fibrosis in 33%, arteritis in 7% and endarteritis obliterans with narrowed lumen in 7% of the patients. No vascular occlusion, ischaemic changes or arteriovenous shunts were observed. These changes were confirmed ultrastructurally. Angiography of the lesion showed a brisk pathological circulation which was more evident in eumycetoma. The vascular Doppler study showed normal blood flow pattern in the affected limb. Regional intra-arterial chemotherapy for mycetoma is suggested as a possible treatment modality.

Internal fistula formation: an unusual complication of mycetoma.

This paper reports 2 hitherto undescribed complications of mycetoma, urinary extravasation and expectoration of mycetoma grains due to cutaneo-urethral and cutaneo-pleuro-bronchial fistulae, respectively. The first patient had an infection with Actinomadura madurae which started in the foot and had spread progressively to involve the whole limb, anterior abdominal wall, perineum and urethra. The second patient had Madurella mycetomatis infection of the hand and, in spite of extensive treatment, the infection had spread to the axilla, chest wall, lung and bronchial tree. Both patients died of the sequelae of these complications. The pathogenesis of these unusual complications is discussed.

Thyroid function in patients with mycetoma.

Thyroid function was assessed in seventy two patients with various types of mycetoma. There was no evidence of clinical or biochemical thyroid dysfunction in these patients. The symptoms encountered in some of the mycetoma patients mimic those of hypothyroidism should be attributed to other factors possibly mental depression and apathy.

The host tissue reaction to Madurella mycetomatis: new classification.

In this prospective study, three types of tissue reaction to mycetoma grains are described. Type I reaction is characterized by the adherence of neutrophils to the surface of the grain leading to its disintegration. In Type II reaction, the fragmented grain and the dead neutrophils are cleared by macrophages and multinucleated giant cells, while in Type III reaction there is a discrete well developed epithelioid granuloma with Langhans's giant cells. The three types of reaction are usually seen in different combinations in the same lesion.

A preliminary study on the ultrastructure of Actinomadura pelletieri and its host tissue reaction.

This is the first report on the fine structure of Actinomadura pelletieri. The in vitro ultrastructure of the organism and that of the grain in vivo were similar. The grain consisted of packed filaments with a faintly electron-dense matrix in between. The individual filament was septate, the cytoplasm was either vacuolated or contained intracellular organelles and the cytoplasmic membrane was adherent to the filamentous wall, which has three electron-dense and two electron-lucent layers. The absence of cement substance around the grain may partially explain the aggressive and destructive behaviour of this organism and may probably also explain its good response to medical treatment. The inflammatory host reaction was similar to that seen with all mycetoma organisms except that there was a massive neutrophil reaction. This was confirmed by electron microscopy.

Post-operative responses of paranasal Aspergillus granuloma to itraconazole.

Paranasal Aspergillus granuloma is an invasive infection, seen mainly in tropical countries, involving the paranasal sinuses, orbit and brain. Previously surgical excision has been followed by a high relapse rate, 80% in some series, and mortality. This study involved the use of post-operative therapy with oral itraconazole in doses of 200-300 mg daily. Twenty-two patients were treated for a mean period of 19.7 weeks. Of 19 patients for whom follow-up data were available, 12 (62%) were rated as being in complete remission in a mean period of 17.2 months after the end of therapy. Only one patient developed progressive disease during itraconazole therapy. No serious adverse effect was seen. Use of itraconazole shows promise as a means of preventing relapse after surgery in this progressive infection.

In vitro studies of antibiotic sensitivities of Streptomyces somaliensis--a cause of human actinomycetoma.

Ten Streptomyces somaliensis strains isolated from mycetoma patients were tested in vitro against 13 antibacterial agents. Rifampicin was the most effective antibiotic in terms of low minimum inhibitory concentration (MIC) followed by erythromycin, tobramycin, fusidic acid and streptomycin sulphate. The S. somaliensis strains were all resistant to trimethoprim, even though the combination of sulphamethoxazole and trimethoprim is commonly used as treatment.

Fatal brain abscesses caused by Ramichloridium obovoideum: report of three cases.

Three cases of brain abscesses caused by Ramichloridium obovoideum are reported. Inspite of adequate antifungal chemotherapy and multiple surgical procedures, all the three patients died of the rupture of a recurrant abscess into the ventricles. Ramichloridium obovoideum was identified as the causative fungus of these brain abscesses. Clinical, histological and mycological features of these cases are described. To our knowledge this is the first time that this fungus is isolated from a brain abscess in Saudi Arabia. The possibility that this life threatening cerebral mycosis may be endemic in this part of the world is considered.

Humoral immune responses to mycetoma organisms: characterization of specific antibodies by the use of enzyme-linked immunosorbent assay and immunoblotting.

Levels of antibodies were determined by enzyme-linked immunosorbent assay (ELISA) in 13 patients with eumycetomas due to Madurella mycetomatis infections. Raised levels of specific IgM were observed in 12 patients, compared with normal human controls. By contrast, low levels of specific IgG were detected in some patients. Specific responses to separated protein antigens were investigated by immunoblotting. Of 10 patients' sera tested, IgM in 2 recognized up to 7 of the blotted antigens between 45 and 84 kDa. Gold-labelled protein A (which predominantly binds to IgG) indicated that sera from 2 patients reacted with at least 6 protein bands with relative molecular masses between 64 and 95. The demonstration of significant IgM levels by ELISA, but few antigenic bands in sera from the same patients by immunoblotting, may point to an antibody response against polysaccharide fungal antigens in mycetoma patients. The use of the ELISA to detect antibodies of different classes and the characterization of their antigenic specificities by immunoblotting may have both diagnostic and prognostic value.

The ultrastructure of actinomycetoma grains caused by Streptomyces somaliensis.

This study compares the in vitro ultrastructure of Streptomyces somaliensis with the grain formation seen in vivo. Cultured forms were cocco-bacillary with thick cell walls and septa. Individual cells stained with PAS.TCH and Alcian blue. Grains were largely composed of an electron-dense fibrillar matrix surrounding clear areas, some of which contained organisms. The ultrastructural appearances of the latter were not significantly different from those seen in vitro although the cell wall was less well defined and did not stain with Alcian blue, and there were more intracellular lipid droplets. Host cells surrounding the mass were incorporated into the structure of the grain, a process associated with cell death. The grains of S. somaliensis appear to be derived from both host and actinomycete elements. Compared to other mycetoma agents the mechanism of grain formation by this organism is distinctive.

Actinomycetoma of the mandibular region causing ankylosis.

A case of mycetoma of the mandibular region caused by Actinomadura madurae is presented. Diagnostic procedures and rationale for treatment are discussed. In tropical and subtropical areas where mycetoma is endemic it should be included in the differential diagnosis of tumours of the facio-maxillary region.

Ultrastructural and immunogenic changes in the formation of mycetoma grains.

The differences in the fine structure and antigenic determinants of mycetoma fungi in the mycelial phase in vitro and in grains in vivo facilitate an interpretation of grain formation. Aggregates of hyphal elements with multiple and thickened walls was a feature of the fungi in vivo. Associated with hyphal wall material, numerous polysaccharide microfibrils were detected in grains of Madurella mycetomatis. These were not seen associated with hyphal elements in vitro and it is suggested that these structures may be concerned with the aggregation of fungal elements in the formation of grains. Antibodies directed against the fungi in vitro were shown, by indirect immunogold labelling, to bind at identical sites in fungal material grown in vitro as in mycetoma grains. However the grain matrix was not labelled, suggesting that part of the structure formed in vivo is composed of modified antigen or is host derived.

Mycetoma of the head and neck.

Cranial mycetoma is not as rare as was believed. In the Sudan, the majority of cases are caused by S. somaliensis; no cases were found to be caused by Nocardia species. Cranial actinomycetoma proved to be potentially fatal and was the most difficult to treat. The best treatment results were achieved in cases of A. madurae infection.