Molecular genotyping of Mycobacterium tuberculosis in Xi'an, China, using MIRU-VNTR typing.

SETTING: The molecular genotyping of Mycobacterium tuberculosis is expected to lead to a better understanding of M. tuberculosis transmission in Xi'an, one of the largest cities in Western China. OBJECTIVE: To characterize the population of M. tuberculosis strains circulating in Xi'an and to evaluate the feasibility of the mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing method. DESIGN: A total of 195 M. tuberculosis clinical isolates from Xi'an were genotyped using spoligotyping and MIRU-VNTR. RESULTS: Thirty-two different genotypes were identified by spoligotyping, with the Beijing family identified as the largest lineage (84.6%), followed by the T1 (3.08%), T2 (1.54%), H3 (1.03%) and U (1.03%) families. The Hunter-Gaston discriminatory index (HGDI) of spoligotyping is low (HGDI = 0.392). In contrast, 15-locus MIRU-VNTR shows a higher discriminatory power (HGDI = 0.998) than 12-locus MIRU-VNTR (HGDI = 0.948). By replacing the ETR C locus with the MIRU39 locus, the discriminatory power of 15-locus MIRU-VNTR is increased to 0.999. CONCLUSIONS: The Beijing family accounts for the vast majority of M. tuberculosis isolates in Xi'an, China. The modified 15-locus MIRU-VNTR showed high HGDI and can be used as a first-line genotyping method in combination with spoligotyping in routine epidemiological investigations in Xi'an, China.

Waterhouse-Friderichsen syndrome after infection with group A streptococcus.

We report a case of Waterhouse-Friderichsen syndrome associated with group A streptococcus (GAS) toxic shock syndrome in a previously healthy man. The patient presented with neck pain and fevers of 2 days' duration. Computed tomography of the neck revealed a mass in the retropharyngeal space, suggesting an abscess. Despite prompt treatment with appropriate antibiotics, the patient experienced a fulminant course and died within 8 hours of presentation. Antemortem blood cultures grew GAS positive for exotoxins A, B, and C. Postmortem examination revealed bilateral adrenal hemorrhage, consistent with Waterhouse-Friderichsen syndrome. Immunohistochemical analysis of the adrenal glands revealed the presence of GAS antigens. However, no disseminated intravascular coagulation was evident. This case demonstrates that adrenal hemorrhage can occur without associated coagulopathy and may result directly from the action of bacterial toxins.

Localization of pigment epithelium derived factor (PEDF) in developing and adult human ocular tissues.

PURPOSE: To localize pigment epithelium-derived factor (PEDF) in developing and adult human ocular tissues. METHODS: PEDF was localized in fetal and adult eyes by immunofluorescence with a polyclonal antibody (pAb) against amino acids 327-343 of PEDF, or a monoclonal antibody (mAb) against the C-terminal 155 amino acids of PEDF. Specificity of the antibodies was documented by Western blotting. PEDF mRNA was localized in adult retina by in situ hybridization. RESULTS: In developing retinas (7.4 to 21.5 fetal weeks, Fwks), pAb anti-PEDF labeled retinal pigment epithelium (RPE) granules, developing cones, some neuroblasts and many cells in the ganglion cell layer (GCL). In adult retinas, pAb anti-PEDF labeled rod and cone cytoplasm and nuclei of rods but not cones. Cells in the INL and GCL, choroid, corneal epithelium and endothelium, and ciliary body were also pAb PEDF-positive. Preadsorption of pAb anti-PEDF with the immunizing peptide blocked specific labeling in retina and other tissues, except for photoreceptor outer segments. In agreement with the immunolocalization with pAb anti-PEDF, in situ hybridization revealed PEDF mRNA in the RPE, photoreceptors, inner nuclear layer cells and ganglion cells in adult retina. In developing retinas 18 Fwks and older, and in adult retinas, mAb anti-PEDF labeled the interphotoreceptor matrix (IPM). Western blots of retina, cornea, and ciliary body/iris with pAb anti-PEDF produced several bands at about 46 kDa. With mAb anti-PEDF, retina produced one band at about 46 kDa; cornea and ciliary body/iris had several bands at about 46 kDa. CONCLUSIONS: PEDF, originally reported as a product of RPE cells, is present in photoreceptors and inner retinal cell types in developing and adult human eyes. Photoreceptors and RPE may secrete PEDF into the IPM.

Prognostic variables for patients with stage III malignant melanoma.

OBJECTIVE: To evaluate the prognostic factors for patients with stage III malignant melanoma and to identify patients at high risk of developing recurrent disease who may benefit from adjuvant therapy. DESIGN: Retrospective study. SETTING: Specialist hospital, USA. SUBJECTS: 130 patients with stage III malignant melanoma (according to the TNM classification), treated at the Roswell Park Cancer Institute between 1970 and 1992. MAIN OUTCOME MEASURES: Survival and prognostic factors on multivariate analysis. RESULTS: Four factors were independent prognostic indicators for patients with stage III malignant melanoma: age >51 years (p = 0.008), >3 involved lymph nodes, (p = 0.03), the site of the primary tumour on head or trunk, (p = 0.007), and the presence of palpable lymph nodes (p = 0.004). CONCLUSION: These prognostic factors help us to stratify patients into low and high-risk groups. High-risk patients may benefit from more aggressive adjuvant therapy in future trials of treatment of melanoma.

Effects of organotypic culture on parasympathetic innervation of guinea pig trachealis.

Nonadrenergic, noncholinergic (NANC) relaxations of airway smooth muscle are thought to be mediated by vasoactive intestinal peptide (VIP) and nitric oxide (NO). Previous studies of the parasympathetic innervation of guinea pig trachealis suggest that the ganglion neurons mediating NANC relaxations but not cholinergic contractions are associated with the esophagus. In this study, the location of the neurons mediating these responses and their neurochemical phenotype was further assessed. Guinea pig tracheas maintained in organotypic culture for 2 days with the adjacent esophagus intact displayed cholinergic contractions and NANC relaxations to electrical field stimulation (EFS) as well as VIP and NO synthase (NOS) nerve fiber densities that were similar to those of control tracheas. By contrast, in tracheas cultured without the esophagus, NANC relaxations to EFS were not observed, and VIP and NOS nerve fiber densities were reduced > 80%. EFS-induced cholinergic contractions were unaffected by esophagus removal. These results provide further evidence that NANC relaxations are mediated by VIP and NO coreleased from noncholinergic parasympathetic nerve endings derived from neurons intrinsic to the esophagus.