Factor XIII-A Val34Leu and Tyr204Phe variants influence clot kinetics in a cohort of South African type 2 diabetes mellitus patients.

Factor XIII, a transglutaminase that plays a crucial role in clot formation, consists of subunits A and B. Single nucleotide polymorphisms in Factor XIII-A have been linked to thrombotic risk. In Type 2 Diabetes mellitus (T2DM), a hypercoagulable state is thought to contribute to the high mortality rate associated with thrombotic diseases. Due to the lack of prevalence data of FXIII-A single nucleotide polymorphisms (SNPs) in T2DM in a South African cohort, this study assessed the prevalence FXIII-A Val34Leu (rs5985) and Tyr204Phe (rs3024477) SNP's and the effect on clot kinetics in T2DM. MATERIALS AND METHODS: A cohort of T2DM patients (n = 100) and race, age and gender matched healthy controls (n = 101) were recruited following ethical approval. Thromboelastography® (TEG®) was used to assess the viscoelastic properties in platelet poor plasma (PPP) in controls (n = 91) and T2DM patients (n = 91) younger than 50 years old. Genomic DNA was isolated from whole blood using the Quick-DNA™ Miniprep Plus Kit and PCR-RFLP was used to genotype each sample for FXIII-A rs5985 and rs3024477 SNPs. RESULTS: TEG® analyses indicated a longer R-time (p < 0.0001) and higher TMRTG (p < 0.0001) in PPP of T2DM patients. Control and T2DM genotype distribution conformed to Hardy-Weinberg equilibrium (p > 0.05). There was a higher prevalence of the wildtype genotype of FXIII-A Tyr204Phe (rs3024477) SNP in T2DM (OR = 0.23, 95% CI = 0.12-0.42, p < 0.0001). The 204Phe variant was more frequent in the Caucasians (OR = 0.39, 95% CI = 0.05-0.33, p < 0.0001). The presence of the 204Phe variant in T2DM affected TMRTG (p = 0.0207). The variant affected R time (p = 0.0432) and TMRTG (p = 0.0209 and p = 0.0207) in controls and T2DM, respectively. CONCLUSION: An inverse association with T2DM and FXIII-A Tyr204Phe was found. A hypo coagulable PPP clot profile was observed in T2DM. A shorter reaction time was observed and but faster rate at which the clot reached maximum strength in both controls and T2DM in the presence of the 204Phe variant.

[Ultrasound-guided puncture : an inexpensive and effective learning model]. / Ultraschallgesteuerte Punktion : Ein kostengünstiges und effektives Lernmodell.

Ultrasound is present in almost any practice and hospital clinic today. In clinical practice, puncture of structures, injections of local anesthetics or glucocorticoids are frequently performed. However, unguided puncture into tissue may have deleterious side-effects if the target is not reached or the drug is not injected properly, such as bleeding, traumatization of delicate structures, glucocorticoid-induced tendon rupture and skin necrosis. Coordination of eye, probe and the manipulation hand needs practice before being used on patients.Therefore, we developed an inexpensive, easy to make and effective learning model for ultrasound-guided puncture and injections. We describe how the model is made and how it can be used to efficiently enhance learning success. It was found that a person unskilled in ultrasonography needs about 40-60 coached punctures in order to confidently hit the target.This model has already been used in our medical education program for rheumatologists, internists, surgeons, orthopedic specialists, anesthetists and general practitioners with great success.

Encapsulation of small spherical liposome into larger flaccid liposome induced by human plasma proteins.

We show that human plasma can induce the encapsulation of small spherical liposomes into larger flaccid liposomes. To explain the observed phenomena, it is proposed that the orientational ordering of charged plasma proteins induces attractive interaction between two like-charged liposome surfaces in close contact. It is observed that the encapsulation of the spherical liposome is possible only if the membrane of the target liposome is flexible enough to adapt its shape to the shape of the spherical liposome. In the theoretical model, the shapes of the two agglutinated liposomes are determined by minimisation of the sum of the adhesion energy and the membrane elastic energy. In the simulations, the membrane of liposomes is considered as an elastic structure and discretised via the finite element method using spring elements. It is shown that the observed agglutination of liposomes and encapsulation of smaller spherical liposomes into larger flaccid liposomes may be explained as a competition between the membrane deformation energy and the membrane adhesion energy.

Polymorphism of bovine MHC class I genes. Joint report of the Fifth International Bovine Lymphocyte Antigen (BoLA) Workshop, Interlaken, Switzerland, 1 August 1992.

The objectives of the Fifth International BoLA Workshop were to: standardize nomenclature, compare typing methods, and characterize BoLA haplotypes. The workshop was based on the distribution of blood samples (cells) from 60 selected cattle to 14 laboratories. Results for the class I (BoLA-A) region are presented in this paper while results for the class II regions are presented in a separate report. Thirty-six of the 50 previously established serological class I specificities were represented in the cell panel. However, only 30 specificities could be confirmed. Two specificities, A16 and A32, were upgraded from provisional, workshop (w) specificities to BoLA-A locus specificities and three new specificities, w51(w28), w52 and w53(w28), were defined. The 39 specificities distinguished 30 class I haplotypes in the 60 animals. Class I isoelectric focusing proved to be a useful adjunct to the serology. Isoelectric focusing confirmed several serologically defined splits and detected splits of A15(A8), A18(A6) and A22(w49) that had not been detected by serology. Subsequently, serological support for splits of A15(A8) and A22(w49) was found.

Oral vaccination of mice against tetanus by use of a live attenuated Salmonella carrier.

A Salmonella typhimurium aroA mutant has been used as a live carrier to immunize mice against tetanus. Plasmid pTETtac4, which expresses a 50-kilodalton fragment of tetanus toxin (fragment C) under the control of the tac promoter, was introduced into SL3261 aroA. When used as a live vaccine and administered orally or intravenously, this strain was able to induce protective immunity in mice against a lethal tetanus toxin challenge. When plasmid pTETtac2, which contains the lacI gene, was used, no immunity was obtained, indicating that the expression of fragment C was repressed in vivo. We believe that this is the first example of a successful oral vaccination that uses an attenuated bacterial carrier to deliver a protective antigen derived from tetanus toxin.

Construction and characterization of vaccine strains of Salmonella harboring mutations in two different aro genes.

Derivatives of the mouse-virulent Salmonella typhimurium strain SL1344 were constructed harboring stable mutations in aroC alone or in aroC and aroA together. Fifty percent lethal doses after intravenous inoculation of the mutants into BALB/c mice were determined, and the mutants were as highly attenuated as were SL1344 aroA derivatives. All aro-dependent derivatives persisted in vivo at similar levels and for similar intervals in the livers and spleens of BALB/c mice infected intravenously. Mice vaccinated orally with 10(10) live organisms of the different derivatives survived and were well protected against oral challenge with virulent S. typhimurium. A Salmonella typhi Ty2 derivative, designated WBL2000, was constructed that harbors well-defined, stable mutations in both aroA and aroC.

Freeze-drying of Anaplasma marginale.

Heparinized whole blood, heavily parasitized with Anaplasma marginale, was collected from 3 splenectomized oxen. Buffered lactose peptone (BLP) was added to equal volumes as a stabilizer and the mixture lyophilized in 2 ml aliquots after rapid freezing. The dried material was reconstituted with 2 ml sterile water and inoculated without delay. The product remained infective for at least 6 months when stored in an ordinary household deep-freeze unit.