Molecular definition and identification of new proteins of Mycobacterium leprae

This report describes N-terminal group analysis of six new proteins isolated from in vivo-grown Mycobacterium leprae, three of which correspond to products of the cysA, ahpC, and rpIL genes, which were recently defined through the M. leprae genome project and which encode a putative sulfate sulfurtransferase, an antioxidant enzyme, and the L7/L12 ribosomal protein, respectively.     

The deleterious effects of human erythropoietin gene driven by the rabbit whey acidic protein gene promoter in transgenic rabbits

Human erythropoietin (EPO) gene and cDNA associated with the rabbit whey acidic protein (WAP) gene promoter were used to tentatively produce the recombinant protein in milk of transgenic mice and rabbits. Several gene constructs showed good efficiency in the mouse mammary cell line HC11. None of them was able to direct the expression of the hormone at a concentration higher than 50 micrograms/mL in mouse and rabbit milk. With one of the construct, the rabbits had an abnormally high amount of red blood cells irrespectively of their sex, they could not reproduce and no milk could be obtained from them. These animals died prematurely. In these animals, the EPO gene was therefore expressed at a low but supraphysiological level in organs other than the mammary gland. These experiments show that transgenic animals obtained with gene constructs which do not contain insulators cannot be used as living fermentors to produce human erythropoietin in their milk at an industrial scale.     

Acute monoarthritis: a practical approach to assessment and treatment

A basic approach to the patient presenting with acute monoarthritis includes a careful history, a physical examination and a selected battery of laboratory tests and radiographs. Because of the possibility of septic joint, rapid assessment and treatment are required. The most common causes of acute monoarthritis are trauma, crystals (gout and pseudogout) and infection. The most important cause of acute monoarthritis is infection, which must be excluded through the use of diagnostic joint aspiration and culture of synovial fluid.     

Cytokinetic effects of cisplatin on diverse human head and neck carcinomas in vitro: dependence on the tumor sensitivity to cisplatin

Studies performed with xenografted human head and neck carcinomas in vivo have demonstrated that the cytokinetic phenomena occurring under the influence of cisplatin closely correlate with the response of the tumors to therapy. The present paper analyses whether this correlation also exists in vitro. Four human head and neck carcinoma cell lines showing different degrees of sensitivity to cisplatin, as determined by the trypan blue exclusion assay, were investigated by flow cytometry at various intervals after administration of cisplatin. Early cell-cycle blockades in the S phase always reflected a high degree of cytostatic potency of cisplatin and were usually succeeded by a pronounced inhibition of tumor cell proliferation and a reduction of cell viability. In the case of a minimal response to therapy and in untreated control cultures of all four tumor lines, the relative number of S-phase cells continuously diminished during the observation period. These findings point to the S-phase blockade as the crucial cytokinetic effect of cisplatin preceding relevant growth reductions. This knowledge might support the development of a drug-response assay that could predict the sensitivity of individual patient tumors in vitro before the beginning of clinical cancer chemotherapy.     

Termination-indicator technique as structured programming tool

The termination-indicator technique is presented, which can be used to solve the class of multi-exit loop problems. A multi-exit loop problem is a programming problem that has several terminating conditions for a loop: either a loop will run through a full cycle or be terminated prematurely when some condition is met. The class of multi-exit loop problems has generated a great deal of controversy on the goto statement in the literature from time to time. The paper illustrates the generality and efficiency of the termination-indicator technique by applying it to the class of multi-exit loop problems and thus puts the goto controversy to rest.     

Extraembryonic expression of the human MHC class I gene HLA-G in transgenic mice. Evidence for a positive regulatory region located 1 kilobase 5' to the start site of transcription

Trophoblast, the only fetal tissue in direct contact with maternal cells, fails to express the polymorphic HLA class I molecules HLA-A and -B, but does express the nonpolymorphic class I molecule HLA-G. It is thought that HLA-G may provide some of the functions of a class I molecule without stimulating maternal immune rejection of the fetal semiallograft. As a first step in identifying the cis-acting DNA regulatory elements involved in the control of class I expression by extraembryonic tissue, several types of transgenic mice were produced. Two HLA-G genomic fragments were used, 5.7 and 6.0 kb in length. These included the entire HLA-G coding region, 1 kb of 3' flanking sequence, and 1.2 or 1.4 kb of 5' flanking sequence, respectively. A hybrid transgene, HLA-A2/G, was produced by replacing the 5' flanking sequence, first exon, and early first intron of HLA-G with the corresponding elements of HLA-A. Comparison of transgene mRNA expression patterns seen in HLA-A2/G and HLA-G transgenic mice suggests that 5' flanking sequences are largely responsible for the differing patterns of expression typical of the classical class I and HLA-G genes. Studies comparing the extraembryonic HLA-G expression levels of founder embryos transgenic for either the 5.7- or 6.0-kb HLA-G transgene showed that the 6.0-kb transgene directed HLA-G expression far more efficiently than did the 5.7-kb HLA-G transgene, producing extraembryonic HLA-G mRNA levels similar to those seen in human extraembryonic tissues. The results of these studies suggest that the 250-bp fragment present at the extreme 5' end of the 6.0-kb HLA-G transgene and absent from the 5.7-kb HLA-G transgene contains an important positive regulatory element. This 250-bp fragment lies further upstream than any of the previously documented class I regulatory regions and may function as a locus control region.     

Effect of different dialysis membranes on platelet function. A tool for biocompatibility evaluation

Intradialytic coagulative and platelet activation, one of the main consequences of blood-membrane contact, was studied in a group of 5 RDT patients with a comparative evaluation of 3 different dialytic membranes: Cuprophan (CU), Polysulfone (PS) and Cellulose Triacetate (CT). Each patient underwent 5 consecutive dialysis sessions with the above mentioned membranes. Intradialytic platelet activation was studied through a morpho-functional evaluation between the mean platelet volume (MPV) and Serotonin (S), beta-Thromboglobulin (beta-TG) and Platelet Factor 4 (PF4) serum levels. These determinations were made before HD (time 0) and after 30', 120', and 240'. We also checked the intradialytic status of thrombogenesis and fibrinolysis determining aPTT, thrombin time, fibrinogen, antithrombin III (AT III), alpha-2 antiplasmin and plasminogen, at the same time intervals. All membranes tested (CU, PS, CT) caused appreciable intradialytic platelet activation, above all after 15' and at the end of dialysis sessions, more marked for CU than PS or CT. In particular MPV showed a decrease throughout the session (-5% at 30' and -9% at 240') while S, beta TG and PF4 peripheral blood levels showed a significant increase at the same intervals with CU membrane. Lastly coagulative and fibrinolytic parameters showed no significant differences among any of the membranes tested.