Genomic diversity of vibrios associated with the Brazilian coral Mussismilia hispida and its sympatric zoanthids (Palythoa caribaeorum, Palythoa variabilis and Zoanthus solanderi).

AIMS: A taxonomic survey of the vibrios associated with the Brazilian endemic coral Mussismilia hispida and the sympatric zoanthids (i.e. Palythoa caribaeorum, Palythoa variabilis and Zoanthus solanderi). METHODS AND RESULTS: Mucus of 54 cnidarian specimens collected in three different places at São Sebastião in two consecutive years (i.e. 2005 and 2006) was used for taxonomic characterization of the cnidarian microbiota. Ninety-eight of the 151 vibrio isolates fell within the vibrio core group according to partial 16S rDNA sequences. We performed the sequencing of recA and pyrH genes of all vibrio isolates. The most abundant taxa belonged to the vibrio core group (Vibrio harveyi, Vibrio rotiferianus, Vibrio campbellii and Vibrio alginolyticus), Vibrio mediterranei (=Vibrio shillonii) and Vibrio chagasii. With the exception of V. chagasii which was found only in the mucus of M. hispida, the other species appeared in different hosts with no evidence for the presence of host-specific clones or species. Using rep-PCR analysis, we observed a high genomic heterogeneity within the vibrios. Each vibrio isolate generated a different rep-PCR fingerprint pattern. There was a complete agreement between the grouping based on rep-PCR and concatenated sequences of pyrH, recA and 16S rDNA, but the pyrH gene has the highest discriminatory power for vibrio species identification. CONCLUSION: The vibrio core group is dominant in the mucus of these cnidarians. There is a tremendous diversity of vibrio lineages within the coral mucus. pyrH gene sequences permit a clear-cut identification of vibrios. SIGNIFICANCE AND IMPACT OF THE STUDY: The taxonomic resolution provided by pyrH (but not recA) appears to be enough for identifying species of vibrios and for disclosing putative new taxa. The vibrio core group appears to be dominant in the mucus of the Brazilian cnidarians. The overrepresentation of these vibrios may reflect as yet unknown ecological functions in the coral holobiont.

Somatic and psychological factors related to the body mass index of patients with anorexia nervosa.

OBJECTIVE: The aim of this study was to examine somatic and psychological factors related to the body mass index (BMI) of anorexia nervosa (AN) patients. METHOD: The analysis was of 24 hospitalized AN patients from the day after admission to the 4th day. The somatic factors analyzed were duration of AN, daily food intake, eating regulatory substances in blood (acylated ghrelin, desacyl ghrelin, leptin), serum cortisol, insulin and estimated creatinine clearance (CCr). The psychological factors analyzed were depression, anxiety, Eating Disorder Inventory (EDI), and hunger/fullness feeling. Measurement of BMI and collection of blood samples were done on the morning after hospitalization. Statistical analysis was by multiple linear regression analysis. RESULTS: BMI showed a reverse correlation with desacyl ghrelin (beta=-0.486, p=0.015) and maturity fears (beta=-0.375, p=0.046), but was not associated with any other factor by multiple regression analysis. CONCLUSION: The results suggest that desacyl ghrelin and maturity fears play important roles in the prolonged malnutrition state seen in AN patients.

Systemic inhibition of tumor growth and tumor metastases by intramuscular administration of the endostatin gene.

Tumors require ongoing angiogenesis to support their growth. Inhibition of angiogenesis by production of angiostatic factors should be a viable approach for cancer gene therapy. Endostatin, a potent angiostatic factor, was expressed in mouse muscle and secreted into the bloodstream for up to 2 weeks after a single intramuscular administration of the endostatin gene. The biological activity of the expressed endostatin was demonstrated by its ability to inhibit systemic angiogenesis. Moreover, the sustained production of endostatin by intramuscular gene therapy inhibited both the growth of primary tumors and the development of metastatic lesions. These results demonstrate the potential utility of intramuscular delivery of an antiangiogenic gene for treatment of disseminated cancers.

Absence of prognostic significance of bcl-2 immunopositivity in non-small cell lung cancer: analysis of 427 cases.

The bcl-2 gene product inhibits apoptosis and is thought to participate in oncogenesis. Association of bcl-2 immunopositivity with improved prognosis of non-small cell lung cancers (NSCLC) is controversial. Although two studies have reported better survival in bcl-2-immunopositive NSCLCs, a third series has contradicted this finding. The authors studied a relatively larger case series involving 427 patients for whom detailed information on long-term follow-up was available to determine the prognostic significance of bcl-2 expression. The study included 252 adenocarcinomas (AC), 111 squamous cell carcinomas (SCC), and 64 large cell carcinomas (LC). After antigen retrieval, sections were immunostained using a monoclonal anti-bcl-2 antibody (1:60, Clone 124, Dako) and the avidin-biotin complex technique. Staining was scored as positive or negative and also on a semiquantitative scale as 0, low (<10%), moderate (10% to 75%), or extensive (>75%). Bcl-2 immunoreactivity was correlated with survival using the actuarial survival method, Kaplan-Meier method, and log-rank test and was not associated with statistically significant differences in survival for NSCLCs (P = .5537). Differences in survival remained insignificant even after NSCLCs were stratified for cell type, stage, or grade, singly or in combination. Therefore, using this method, bcl-2 immunopositivity does not appear to act as an independent prognostic indicator in NSCLCs.

p53 and MDM2 immunostaining in pulmonary blastomas and bronchogenic carcinomas.

Pulmonary blastomas (PBs) are rare primary malignancies that include adult types: biphasic pulmonary blastoma (BPB) and well-differentiated fetal adenocarcinoma (WDFA); and childhood type: pleuropulmonary blastoma (PPB). Their pathogenesis and relationship to bronchogenic carcinoma (BCA) are controversial. To determine whether or not PB share molecular pathological features with BCA, the authors immunostained three BPB, three WDFA, three PPB, and 80 standard BCA for p53 protein and MDM2 protein, gene products believed to be significant in the pathogenesis of BCA. Paraffin-embedded tissue sections were immunostained with monoclonal antibody to p53 and MDM2 proteins. Strong intranuclear staining in greater than 10% of cells was considered positive. Three (50%) BPB and WDFA stained for p53 and five (83%) for MDM2. None of the PPB stained for p53, and one PPB did not stain for either p53 or MDM2. Five of six adult type PB occurred in smokers, whereas none of the PPB was associated with smoking. Seventy-five (94%) of the BCA stained for MDM2 and 46 (61%) for p53. Immunostaining patterns for p53 and MDM2 in adult types of PB, and not PPB, appear similar to those for BCA. This may suggest that adult type PB, but not childhood PB, have a similar pathogenesis to BCA.

Fatal Pneumocystis carinii pneumonia in HIV-seropositive infants in Harare, Zimbabwe.

Lung biopsies taken post mortem from 24 HIV-seropositive children who died of pneumonia in Harare Hospital (Zimbabwe) during 1995 were examined for pathogens using histology, culture, microscopy and polymerase chain reaction (PCR). Pneumocystis carinii was detected in 16 (67%) children, in 5 of whom bacterial pathogens were also detected. There were 2 cases of cytomegalovirus infection. On the basis of histology and PCR, none of the children had tuberculosis. These data add to the evidence that P. carinii pneumonia may be a significant cause of death in HIV-infected children in southern Africa. Policies on treatment for severe pneumonia, and on prophylaxis for children born to HIV-seropositive mothers need to be re-examined.

p53 gene product in pleural effusions. Practical use in distinguishing benign from malignant cells.

OBJECTIVE: To determine the utility of positive p53 immunostaining as an adjunct in the diagnosis of malignancy in pleural effusions, we reviewed 103 effusions representing the typical range of diagnoses encountered in the evaluation of pleural fluid cytology. STUDY DESIGN: Immunohistochemistry was performed on paraffin-embedded cell blocks using a monoclonal antibody to the p53 suppressor gene product clone BP53-12 and a standard avidin-biotin complex technique with a citrate buffer antigen retrieval solution. RESULTS: Forty-one of 75 effusions with an unequivocal cytologic diagnosis of malignancy were immunopositive for p53 protein (55%). One of nine effusions cytologically interpreted as showing reactive mesothelial cells showed immunopositivity; that case was subsequently diagnosed as a mesothelioma on pleural biopsy. Nineteen cases were interpreted as suspicious for malignancy. Of these, 16 were negative, and 3 were positive for p53 protein. Of the three positive cases, two showed the presence of non-small cell and poorly differentiated large cell carcinoma. CONCLUSION: These findings suggest that p53 protein immunostaining is relatively sensitive and highly specific in differentiating benign mesothelial cells from malignant cells in pleural effusions. While negative p53 protein immunostaining does not exclude malignancy, positive staining in reactive or suspicious cells warrants further diagnostic evaluation of the patient.

[A giant chondromyxoid fibroma originated from the right orbital roof.--A case report--].

The authors reported a case of giant chondromyxoid fibroma of the right anterior cranial fossa, arising from the right orbital lamina of frontal bone. A fifteen-year-old boy was admitted because of a recent history of the right exophthalmus and headache. Neurological examination was essentially negative except papilledema in the both optic fundi and the right olfactory disturbance. Skull plain x-ray films showed the bony destruction of the right supraorbital bone and the some of abnormal calcification in the right anterior cranial fossa. CT scan showed cystic low density spots surrounded by irregular ring-like high density areas in the right anterior cranial fossa. Operation was performed on two stages and the tumor was removed totally. The tumor was arising from the orbital lamina of the frontal bone. The size of resected tumor was 7x5x4 cm. The pathological examination confirmed the diagnosis of chondromyxoid fibroma. Postoperatively, the patient is fully schooling without any disturbance 2 years and 7 months after the discharge. In Japan, two cases of intracranial chondromyxoid fibroma have been reported in literature. The authors discussed the histology of chondromyxoid fibroma and the genesis of the membraneous bone origin of the intracranial chondromatous tumor.

[Vasculo-Behçet's disease with superior sagittal sinus thrombosis--case report (author's transl)].

The patient is a 30-year-old man who has suffered from recurrent attacks of tonsilitis, oral aphthae and scrotal ulcerations, erythema nodosum and thrombophlebitis. In April, 1980, he gradually developed headache and visual disturbance. On April 14, 1980, he was pointed out remarked bilateral choked disc by an ophthalmologist and then admitted to the Miyazaki Medical College Hospital. On admission to our service, he showed atypical symptoms of Behçet's disease, namely, oral aphthae and scrotal ulcerations, erythema nodosum and bilateral choked disc. Laboratory data demonstrated hyperimmunoglobulinemia, increased clotting factors and decreased fibrinolytic activity. Immunogenetically, HLA BW51 type was demonstrated. The angiograms showed complete obstructions of the superior sagittal sinus and the common trunk of the femoral artery. Histological examination of the skin lesion demonstrated atypical chronic inflammation and thrombophlebitis. A diagnosis of atypical Vasculo-Behçet's disease was made. The response to the steroid therapy was dramatic, though the fibrinolytic drugs, anticoagulants and vasodilators were not effective. Thrombophlebitis is a well recognized complication of Behçet's disease occurring in major vessels, however thrombosis of the dural sinus has rarely reported. This case may be the first one which had superior sagittal sinus thrombosis with Vasculo-Behçet's disease in literature. We discussed the mechanism of the thrombogenesis, the relationship to HLA, the coexistence of Neuro-Behçet's disease and the therapy of Vasculo-Behçet's disease.

Direct formation of angiotensin II without renin or converting enzyme in the ischemic dog heart.

An in vivo study was done to determine whether angiotensin II is directly formed by kallikrein or kallikrein-like proteases. Dogs were bilaterally nephrectomized 24 hours before ligation of the coronary artery. This acute coronary artery occlusion led to a regionally increased acidic state in the ischemic tissue and resulted in an elevation of immunoreactive angiotensin II (IR-Angiotensin II) levels in the coronary sinus blood, but not in systemic aortic blood. Elevation of the IR-Angiotensin II level was specifically inhibited by aprotinin, a kallikrein inhibitor. It was not affected by either captopril, a potent angiotensin converting enzyme inhibitor, or by pepstatin, a renin inhibitor. The concentrations of immunoreactive angiotensin I (IR-Angiotensin I), plasma renin activity and angiotensin converting enzyme activity remained unaltered in the presence of coronary artery occlusion. These results suggest that IR-Angiotensin II in the ischemic heart may be generated directly by kallikrein or kallikrein-like proteases, independently of the systemic renin angiotensin system.

Prognostic implications of calbindin-D28k expression in lung cancer: analysis of 452 cases.

Calbindin D28k (Ca-D28k) acts as a buffering system to maintain cellular calcium homeostasis and is thought to play a role in inhibiting apoptosis. The goals of this study were to assess CA-D28k expression in lung carcinomas and to correlate these results with patient survival. A total of 452 lung carcinomas were immunostained with a monoclonal antibody specific for Ca-D28K using an avidin-biotin peroxidase technique. The number of cells with nuclear staining was graded semiquantitatively into one of five groups: 0, fewer than 10%, 10 to 25%, more than 25 to 50%, more than 50 to 75%, and more than 75%. Results were correlated with patient survival using Kaplan-Meier survival curves. A total of 335 of 452 (74%) lung carcinomas were positive for Ca-D28k. There was no statistically significant difference in the prevalence of Ca-D28k expression in tumors of different histologic type. Kaplan-Meier survival analysis revealed that for patients with adenocarcinoma, those with Ca-D28k-positive tumors had a better overall survival than patients with Ca-D28k-negative tumors (P = .036). This difference was also significant for patients with Stages I and II adenocarcinomas (P = .033). No statistically significant difference in prognosis was observed for patients with Stages III and IV adenocarcinomas or for patients with other lung carcinoma types of varying stage. Ca-D28k is commonly expressed in lung carcinomas of all histologic types. For patients with localized adenocarcinoma of the lung, Ca-D28k expression correlated with improved survival. No correlation between Ca-D28k expression and patient survival was found for disseminated adenocarcinoma and for other histologic types of lung carcinoma.

Synthesis and antitumor activity of novel dolastatin 10 analogs.

Dolastatin 10 (1) is a potent antineoplastic pentapeptide. Novel dolastatin 10 analogs each modified at one of the constituent amino acid derivatives, were synthesized and their antitumor activity was evaluated against P388 leukemia in mice. The structural requirements for antitumor activity are discussed. Some of the analogs, 31c, 35c, 38b, and 50c showed excellent activity in vivo. Highly active 50c, which lacks the thiazole group of 1, was selected for further development as an antitumor agent.

The influence of glycine and related compounds on spinal cord injury-induced spasticity.

Spasticity is a frequent and complex sequel to spinal cord injury. The neurochemical basis for the origin of spasticity is largely unknown. Glycine is among the most abundant neurotransmitters in the spinal cord. However, the role of glycine and related compounds in spasticity have received little attention. An ischemic spinal cord injury was created in rabbits, by an intraaortic balloon occlusion technique, which produced lower limb spasticity. A catheter was inserted into the cisterna magna and the spinal cord was bathed with 100 microM solutions of glycine, strychnine, D-serine, beta-alanine, MK-801, or artificial CSF for 4 hours at a rate of 10 microliters/min. H-reflexes were monitored before and during infusion by stimulating the posterior tibial nerve and recording from the plantar surface of the foot. Glycine, D-serine, and MK-801 depressed the H wave, strychnine produced a heightened H wave, and beta-alanine caused no significant changes. These results indicate that glycine and related compounds may influence spasticity.