Isolation and Characterization of Mycoplasma conjunctivae sp. n. from Sheep and Goats with Keratoconjunctivitis

Fourteen conjunctival strains of mycoplasma were isolated from infected sheep and goats in two separate naturally occurring outbreaks of keratoconjunctivitis (pink-eye). The biological and serological properties and cell protein patterns of these strains are presented in this report. These strains were related to each other and were unrelated to 40 recognized Mycoplasma and Acholeplasma species and serotypes. We are proposing that this group of mycoplasmas be named M. conjunctivae. The role of M. conjunctivae in the etiology of keratoconjunctivitis in sheep and goats remains to be determined.     

Removal of the intruding Miragel's scleral buckle by pars plana ultrasonic fragmentation

PURPOSE: To introduce a surgical approach for removing intruded scleral buckle resulting from Miragel's implant. DESIGN: Interventional case report. METHODS: After cutting and releasing scleral band externally, a pars plana Fragmatome was used to break up and aspirate pieces of the Miragel's implant that had grown into the eye, filling the vitreous cavity. RESULTS: The intruding Miragel's buckle was shaved to the plane of the retina without any complication. The retina was completely attached. CONCLUSION: Use of ultrasonic fragmentation by a pars plana approach was effective in removing the intavitreally expanded buckling element without harming the retina.     

Correlation of in vitro cytotoxicity with paracellular permeability in Caco-2 cells.

This in vitro study aims to develop a cell culture model that compares paracellular permeability (PP) with acute cytotoxicity (AC). Caco-2 cells were seeded in 96-well plates and on polycarbonate filter inserts. Confluent monolayers were exposed to increasing concentrations of 20 reference chemicals for 24-h and 72-h. Cytotoxicity was determined using MTT and NRU cell viability assays in 96-well plates. PP was measured using transepithelial electrical resistance (TEER) measurements, as well as passage of lucifer yellow (LY), [3H]-mannitol (both low mw indicators), and FITC-dextran (higher mw indicator) in culture inserts. Inhibitory concentrations 50% (IC50s) suggest that there were good correlations between 24-h and 72-h exposures. NRU IC50 values correlated better with TEER, which is consistent with the Registry of Cytotoxicity (RC; ICCVAM) database report. Both cell viability assays indicate that cytotoxicity occurs before TEER is compromised. In addition, 24-h and 72-h NRU assays, and 72-h TEER measurements, displayed the highest correlations with established rodent LD50s. PP experiments showed that passive paracellular transport of the tight junction markers, especially [3H]-mannitol, correlates with the IC50s determined with the viability assays and TEER measurements. Our AC/TEER/PP model thus allows for the differentiation between the concentrations necessary for AC and those needed to interfere with PP. We propose that the in vitro AC, TEER and PP results be used to compute a formula which can "normalize" and improve the predictive ability of in vitro acute cytotoxicity assays for in vivo lethality.     

Urogenital challenge of primate species with Mycoplasma genitalium and characteristics of infection induced in chimpanzees

Eighteen male and eight female primates, representing five subhuman species, were inoculated urogenitally with Mycoplasma genitalium, a microorganism recovered from men with nongonococcal urethritis. Male rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) monkeys apparently were resistant. Female squirrel monkeys (Saimiri sciureus) and female tamarins (Saguinus mystax) exhibited low-level, genital-tract infections. Male chimpanzees (Pan troglodytes) developed an obvious genital-tract infection, with some shedding organisms for 21 weeks. M. genitalium was recovered from the blood of two of the male chimpanzees, usually when large numbers of organisms were in the urethra. Female chimpanzees generally shed organisms for 12-15 weeks. Most chimpanzees colonized with the organism exhibited increased numbers of polymorphonuclear leukocytes in the genital tract and developed a significant antibody response. The results offer substantial evidence for the pathogenicity of M. genitalium for the urogenital tract of higher primates and suggest the microorganism may have a role in human genital-tract infections.     

High prevalence of the G101W germline mutation in the CDKN2A (P16(ink4a)) gene in 62 Italian malignant melanoma families.

CDKN2A germline mutation frequency estimates are commonly based on families with several melanoma cases. When we started counseling in a research setting on gene susceptibility analysis in northern and central Italy, however, we mostly found small families with few cases. Here we briefly characterize those kindred, estimate CDKN2A/CDK4 mutation test yields, and provide indications on the possibility of implementing formal DNA testing for melanoma-prone families in Italy. In September 1995 we started genetic counseling in a research setting at our Medical Genetics Center. Screening for CDKN2A/CDK4 mutations was performed on families with two melanoma patients, one of whom was younger than 50 years at onset, the other complying with one of the following: 1) being a first-degree relative, 2) having an additional relative with pancreatic cancer, or 3) having multiple primary melanomas. Sixty-two of 67 (80%) melanoma cases met our criteria. Four previously described CDKN2A mutations (G101W, R24P, V126D, and N71S) were found in 21 of the 62 families (34%) with a high prevalence of G101W (18/21). The percentage of families with two melanoma cases/family harboring a mutation was low (7%, 2/27), but rose to 45% (9/20) if one of the melanoma patients carried multiple melanomas or if pancreatic cancer was present in that family. In the 15 families with three melanoma cases the presence of a mutation was higher (67%, 10/15) and reached 100% in the 4 families with four or more melanoma cases. Our results suggest that CDKN2A/CDK4 counseling-based mutational analysis may be reasonably efficient also for families with two melanoma cases, if one patient carries multiple melanomas or if pancreatic cancer is present in the family.     

Therapy of refractory/relapsed acute myeloid leukemia and blast crisis of chronic myeloid leukemia with the combination of cytosine arabinoside,tetrahydrouridine, and carboplatin

Summary Eight patients, of whom four had acute myeloid leukemia (AML) and four had chronic myeloid leukemia (CML) blast crisis, were treated with a combination of cytosine arabinoside (ARA-C: 1,600 mg/m² in three patients, 1,200 mg/m² in five patients), tetrahydrouridine (THU: 2,800 mg/m² in two patients, 2,646 mg/m² in one patient, 2,100 mg/m² in five patients), and carboplatin (900 mg/m² in four patients, 720 mg/m² in one patient, 450 mg/m² in three patients). As a result of this treatment, five of the eight patients became aplastic. Two of the four patients with CML blast crisis reverted to the chronic phase and two of the four patients with acute nonlymphocytic leukemia (ANLL) attained a remission (one partial remission and one complete remission). The major toxicities included myelosuppression, unacceptable hepatotoxicity, and diarrhea. Pharmacokinetics studies revealed that the addition of carboplatin did not significantly change the disposition of ARA-C. ARA-C levels were not significantly changed in comparison with those obtained in a prior study of ARA-C with THU (ARA-C plasma levels at 3 h, 2630±1170 ng/ml).Supported by the Don Monti Memorial Research Foundation     

Pars plana vitrectomy for persistent, visually significant vitreous opacities

PURPOSE: To evaluate the role of vitrectomy in patients with persistent, visually disabling vitreous opacities. METHODS: Six consecutive eyes of five men (age 58-66 years) with pseudophakia or aphakia and vitreous opacities resulting in visual symptoms for more than 1 year that underwent vitrectomy were retrospectively reviewed. Postoperative questionnaires regarding functional performance and quality-of-life issues were completed by the participants to assess subjective patient satisfaction. RESULTS: Postoperative Snellen visual acuity was improved or equal to preoperative acuity in all cases (8-44 month follow-up) and there were no surgical complications. All patients expressed high satisfaction with overall visual function. Analysis of the National Eye Institute Visual Function Questionnaire-39 indicated that general vision, near activities, distance activities, mental health, role difficulties, and peripheral vision were significantly improved (P < 0.05) following surgical intervention. CONCLUSIONS: Vitrectomy may be indicated in a select group of patients with visually disabling vitreous floaters, although objective assessment of visual dysfunction from vitreous floaters requires further evaluation.