The identification of mycobacteria from solid media and directly from VersaTREK Myco bottles using the Sherlock Mycobacteria Identification HPLC system.

The Sherlock Mycobacteria Identification HPLC system correctly identified to the species level 61 (67.8%) of 90 isolates growing on solid media, and 73 (45.3%) of 161 isolates directly from positive VersaTREK Myco bottles. When these data were re-analysed with a revised database, correct identifications increased to 91.1% and 83.2%, respectively. All Mycobacterium tuberculosis isolates were identified correctly, regardless of the inoculum source or database used. The use of the revised database with isolates obtained directly from positive VersaTREK Myco bottles allows the identification of most isolates within clinically relevant time-frames.

[Open plastic correction of postoperative hernia].

Incisional hernia is a frequent complication of abdominal surgery. The use of prosthetic materials in open incisional hernia substantially repairs diminished rate of reherniation. The object of this study was the retrospective review of the first seventy open incisional hernia repairs performed at the Tbilisi Municipal Thoraco-Abdominal Clinic. Over a 3-year period 70 open abdominal wall incisional hernia repairs were performed using the retromuscular, preperitoneal, intraperitoneal and prefascial techniques. The prosthetic materials used were polypropylene, prolene and ePTFE. Recurrence rates were 2.8%, both patients had wound infections that required removal of prosthesis. There were no gastrointestinal complications, fistulas or death. Seromas occurred in eight patients (11.4%). Mean length of stay was 5.4 day. In these series open mesh repair had excellent results with minimal morbidity and hospital stay.

Antitumor effects of ajulemic acid (CT3), a synthetic non-psychoactive cannabinoid.

One of the endogenous transformation products of tetrahydrocannabinol (THC) is THC-11-oic acid, and ajulemic acid (AJA; dimethylheptyl-THC-11-oic acid) is a side-chain synthetic analog of THC-11-oic acid. In preclinical studies, AJA has been found to be a potent anti-inflammatory agent without psychoactive properties. Based on recent reports suggesting antitumor effects of cannabinoids (CBs), we assessed the potential of AJA as an antitumor agent. AJA proved to be approximately one-half as potent as THC in inhibiting tumor growth in vitro against a variety of neoplastic cell lines. However, its in vitro effects lasted longer. The antitumor effect was stereospecific, suggesting receptor mediation. Unlike THC, however, whose effect was blocked by both CB(1) and CB(2) receptor antagonists, the effect of AJA was inhibited by only the CB(2) antagonist. Additionally, incubation of C6 glioma cells with AJA resulted in the formation of lipid droplets, the number of which increased over time; this effect was noted to a much greater extent after AJA than after THC and was not seen in WI-38 cells, a human normal fibroblast cell line. Analysis of incorporation of radiolabeled fatty acids revealed a marked accumulation of triglycerides in AJA-treated cells at concentrations that produced tumor growth inhibition. Finally, AJA, administered p.o. to nude mice at a dosage several orders of magnitude below that which produces toxicity, inhibited the growth of subcutaneously implanted U87 human glioma cells modestly but significantly. We conclude that AJA acts to produce significant antitumor activity and effects its actions primarily via CB(2) receptors. Its very favorable toxicity profile, including lack of psychoactivity, makes it suitable for chronic usage. Further studies are warranted to determine its optimal role as an antitumor agent.

Bone marrow accessory cells regulate human bone precursor cell development.

OBJECTIVE: Much remains to be learned about the intimate relationship between bone marrow and its surrounding tissue: the bone. We hypothesized that bone marrow accessory cell populations might regulate the development of human bone precursor cells. MATERIALS AND METHODS: We used immunologic phenotyping, and isolation methods to fractionate subpopulations of nonadherent, low-density (NALD) human bone marrow cells. These cells were examined for their ability to support the serum-free survival, proliferation, and expression of bone proteins by highly purified populations of human bone precursor cells. Quantitative assessment of the accessory cell populations as well as human bone precursor cells phenotype was performed using multiparameter flow cytometry. Bone protein expression was evaluated by immunocytochemistry, Western analysis, and enzymatic analysis (for alkaline phosphatase activity). RESULTS: Human bone marrow contains a cell population that stimulates the development of purified bone precursor cells. Feeder-layer studies demonstrate that these osteopoietic accessory cells (OACs) do not require cell-cell interaction to promote bone precursor cell development but, rather, produce soluble molecules responsible for their effects. Flow cytometric analyses reveal that bone marrow derived B cells, T cells, macrophages, natural killer cells, and endothelial cells do not produce this stimulatory factor. The (growth) factor cannot be replaced by addition of exogenous cytokines. The isolation of human transforming growth factor beta receptor type II (TGF-betaRII)-positive cells increases OAC-specific activity in bone cell ex vivo expansion cultures. Moreover, isolation of OAC bone marrow cells characterized by high TGF-betaRII expression, relatively low cellular complexity, and small size yields a population that is highly enriched for OACs. CONCLUSION: We conclude that human bone marrow contains a population of OACs that are an obligate requirement for the early phases of bone cell development ex vivo.

Thrombopoietin requires additional megakaryocyte-active cytokines for optimal ex vivo expansion of megakaryocyte precursor cells.

Little is known concerning the interaction of thrombopoietin (TPO) with other megakaryocyte-active cytokines in directing the early events of megakaryocyte development. Culture of CD34(+) cells in interleukins (IL) -1, -6, -11, plus stem cell factor (SCF; S) results in a 10- to 12-fold expansion in total cell numbers, whereas total CD41(+) megakaryocytes are expanded approximately 120-fold over input levels. Addition of TPO to IL-1, -6, -11, S generates a biphasic proliferation of CD41(+) cells, accelerates their rate of production, and results in an ex vivo expansion of more than 200-fold. The addition of Flt-3 ligand (FL) increases CD41+ cell expansion to approximately 380-fold over input levels. In the absence of TPO, approximately 95% of the expanded cells show the phenotype of promegakaryoblasts; TPO and/or FL addition increases CD41 antigen density and ploidy in a subpopulation of promegakaryoblasts. A moderate (approximately sevenfold) expansion of megakaryocyte progenitor cells (colony-forming unit-megakaryocyte) occurs in the presence of IL-1, -6, -11, S, and the addition of TPO to this cocktail yields an approximately 17-fold expansion. We conclude that early proliferative events in megakaryocyte development in vitro are regulated by multiple cytokines, and that TPO markedly affects these early developmental steps. However, by itself, TPO is neither necessary nor sufficient to generate a full proliferative/maturational in vitro response within the megakaryocyte compartment. TPO clearly affects terminal differentiation and the development of (some) high-ploidy human megakaryocytes. However, its limited in vitro actions on human cell polyploidization suggest that additional megakaryocyte-active cytokines or other signals are essential for the maximal development of human megakaryocytes.

Human hematopoietic progenitor cell isolation based on galactose-specific cell surface binding.

The ability to isolate functional populations of hematopoietic progenitor cells is important to the process of hematopoietic cell transplantation and to the understanding of hematopoietic cell biology in health and disease. We show that a subpopulation of human bone marrow hematopoietic cells bearing the pan-hematopoietic antigen CD34 also binds galactose-conjugated proteins. This lectin-positive sub-population represents approximately 0.1 to 0.5% of the total bone marrow cells, and contains 100% of the hematopoietic progenitor cells. The galactose-binding lectin on these cells is specific for this sugar. Additionally, highly proliferative hematopoietic progenitor cells with very primitive phenotypes, including a newly identified progenitor cell that produces multiple lineages, express this lectin.

[Extrathoracic surgical emergencies in hospitalized patients with bronchopulmonary diseases. Analysis of the operative risk]. / Le urgenze chirurgiche extratoraciche nel broncopneumopatico ospedalizzato. Analisi del rischio operatorio.

An experience of surgical non-thoracic emergencies in patients admitted for chronic lung disease is herein presented. Fifty-four patients out of 10457 admitted in the four Departments of Pneumology of the Binaghi Hospital (Cagliari) between 1-1-1985 and 31-3-1993, were referred to our Department of General Surgery due to non-thoracic surgical emergencies. There was a considerable delay in the referral (only 25% of patients within 12 hours from the onset of symptoms): indeed predominant respiratory symptoms, hypoxia and hypercapnia made these patients no responsive to symptoms of surgical emergency. Surgical emergencies in causal correlation with respiratory disease (intestinal occlusion due to abdominal metastases of lung carcinoma, complicated peptic ulcer) had the worst prognosis (mortality: 52.9%). Those in chance connection, such as acute limb ischemia and preexisting abdominal disease, had a less adverse outcome. Mortality, however, was 37.5%: this datum outlines the role of chronic lung disease in defining operative risk. The authors call attention to three groups of observed patients: 1) three patients were operated on for intestinal occlusion due to unrecognized abdominal neoplasia, that showed itself in the course of hospitalization in the Department of Pneumology for lung metastases; 2) in 3 cases symptoms and signs of acute abdomen were observed without abdominal disease. The cause of acute pseudoabdomen was diaphragmatic pleural or basal pulmonary inflammation; 3) the eight patients with pulmonary embolism were all admitted in the Department of Pneumology with a wrong diagnosis of bronchopneumonia.

[Splenic metastases of pulmonary carcinoma. Apropos of a clinical case]. / Metastasi spleniche da carcinoma polmonare. A proposito di un caso clinico.

A case of single splenic metastasis from large cell anaplastic carcinoma of the lung is described. The patient first underwent pulmonary lobectomy and then splenectomy. He is still alive 42 months after the first operation and 41 after the second. The case herein described is rather uncommon, due to the rarity of single splenic metastases, and the long survival of the patient. The latter point strongly suggests an aggressive surgical approach to single metastases, even if primary tumor has extremely poor prognosis.