Anthropometric dimensions provide reliable estimates of abdominal adiposity: A validation study.

Abdominal fat accumulation is a major risk factor for cardiometabolic morbidity and mortality. The purpose of the study is to assess the possibility of developing accurate estimation equations based on body measurements to determine total abdominal (TFA), subcutaneous (SFA) and visceral fat area (VFA). Hungarian volunteers (n=198) aged between 20 and 81 years were enrolled in the study, which was conducted between July and November 2014. All persons underwent anthropometric measurements and computer tomographic (CT) scanning. Sex-specific multiple linear regression analyses were conducted in a subgroup of 98 participants to generate estimation models, then Bland-Altman's analyses were applied in the cross-validation group to compare their predictive efficiency. The variables best predicting VFA were hip circumference, calf circumference and waist-to-hip ratio (WHR) for males (R2=0.713; SEE=5602.1mm2) and sagittal abdominal diameter (SAD), WHR, thigh circumference and triceps skinfold for females (R2=0.845; SEE=3835.6mm2). The SFA prediction equation included SAD, thigh circumference and abdominal skinfold for males (R2=0.848; SEE=4124.1mm2), body mass index and thigh circumference for females (R2=0.861; SEE=5049.7mm2). Prediction accuracy was the highest in the case of TFA: hip circumference and WHR for males (R2=0.910; SEE=5637.2mm2), SAD, thigh circumference and abdominal skinfold for females (R2=0.915; SEE=6197.5mm2) were used in the equations. The results suggested that deviations in the predictions were independent of the amount of adipose tissue. Estimation of abdominal fat depots based on anthropometric traits could provide a cheap, reliable method in epidemiologic research and public health screening to evaluate the risk of cardiometabolic events.

Unusual spinal tuberculosis in an Avar Age skeleton (Csongrád-Felgyo, Ürmös-tanya, Hungary): A morphological and biomolecular study.

The paleopathological analysis of a well-preserved young adult female skeleton from the AD 7-8th century (Avar Age) in Hungary revealed multiple lytic lesions in all of the thoracic and lumbar vertebral bodies. The lesions were characterized by smooth marginal zones and space-occupying mass appearance. The considerable loss of spongy bone in the thoracolumbar vertebrae resulted in angular deformity and fusion, characteristic of the healing stage of TB. Osteolytic lesions were also observed on the vertebral processes, ribs and sternum. On the endocranial surface, abnormal blood vessel impressions were revealed, indicating some kind of meningitis. The X-ray and CT analysis of the affected bones detected abnormal structures and cystic zones of destruction. The lesions were however not always bordered by areas of increased density, which is typical in cystic TB. Vertebral remains were also subjected to biomolecular analysis in two different laboratories, which attested the presence of Mycobacterium tuberculosis complex (MTBC) DNA and supported the paleopathological diagnosis of TB. Spoligotyping analysis confirmed the presence of MTBC DNA and more specifically an infection caused by bacteria belonging to the M. tuberculosis lineage. This case study provides new data for the paleoepidemiology of TB in this geographical area and historical period, and draws attention to the great variability of TB lesions in the human skeleton.

Increased expression of the blood group-related Lewis Y antigen on synovial fluid granulocytes of patients with arthritic joint diseases.

OBJECTIVE: To evaluate the expression of the carbohydrate structures Lewis Y (LeY), sialyl-LeX (sLeX) and Lewis X (LeX) on paired peripheral blood (PB) and synovial fluid (SF) granulocytes in patients with arthritic diseases. METHODS: Ten patients with rheumatoid arthritis (RA), seven patients with spondyloarthritis (SA) and eight patients with osteoarthritis (OA) were studied. Granulocyte expression of the Le oligosaccharides was analysed by fluorescence-activated cell sorting. RESULTS: SF granulocytes of patients with RA, SA and OA expressed higher levels of the LeY oligosaccharide than PB granulocytes. Increases in LeY on SF granulocytes were similar in all three underlying diseases. No differences in the expression of the Le antigens were detected between PB granulocytes of patients and healthy individuals. Expression of sLeX and LeX showed no variation between SF and PB neutrophils. CONCLUSION: The selective increase in LeY antigen on SF granulocytes in RA, SA and OA suggests a role of the LeY oligosaccharide in granulocyte traffic and inflammatory responses.

Molecular evidence for different stages of tuberculosis in ancient bone samples from Hungary.

This paleomicrobiologic study was conducted on osseous tissue specimens from ancient Hungarian skeletal samples from the 7-8th and the 17th centuries AD with typical macromorphologic evidence of osseous tuberculosis (n = 3), morphologic alterations probably due to tuberculosis (n = 6), or with nontypical osseous changes of vertebral bodies suggestive of inflammatory reaction (n = 5). From these bone samples, DNA was extracted and amplified by polymerase chain reaction (PCR) by using various primer pairs recognizing DNA segments of different mycobacterial species. To confirm specificity of the analysis, the amplification products of several samples were subjected to restriction enzyme digestion and/or direct sequencing. Of the analyzed 14 cases, 8 were unambiguously positive for mycobacterial DNA of the Mycobacterium tuberculosis complex, as shown by the amplification of the IS6110 sequence. In 13 cases we found a PCR product with primers specific for the 65-kDa antigen gene, including 2 cases without genomic DNA. We conclude that the application of other mycobacterial DNA primers may reveal contamination of bones with atypical saprophytic mycobacteria. A positive result for typical mycobacteria was seen in 2 of 3 cases with typical morphologic signs of tuberculosis and amplifiable DNA, in 3 of 6 probable cases, but also in 3 of 6 cases with nontypical bone changes. This indicates that minor osseous reactions of the surface of vertebral bodies may be due-at least in several cases-to infections with bacteria of the M. tuberculosis complex. In these cases the disease may have proceeded rapidly, and the morphologic osseous changes may represent "early" stages of tuberculous infection of the vertebrae.

Activation-dependent expression of the blood group-related lewis Y antigen on peripheral blood granulocytes.

The expression of the difucosyl-lactosamine type 2 oligosaccharide Lewis Y (LeY) on peripheral blood cells was investigated. As assessed by the reactivity with the mouse anti-LeY monoclonal antibody (mAb) ABL 364 among circulating blood cells, the expression of the LeY oligosaccharide was uniquely restricted to granulocytes. Although the density of LeY expressed on resting granulocytes was weak, in vitro activation of granulocytes with fMLP induced a rapid and pronounced increase in granulocyte LeY expression. Analysis of CEA-related glycoproteins immunoprecipitated with anti-CD66 mAbs followed by immunoblotting with mAb ABL 364 showed that granulocyte LeY is attached to members of the CD66 cluster, in particular to the 160/90 kD glycoprotein recognized by anti-CD66 mAb CBL/gran 10. The activation-associated increase in LeY attached to CD66 adhesion molecules implicates a role of the LeY determinant in the cytoadhesive properties of granulocytes.

Detection of leprosy in ancient human skeletal remains by molecular identification of Mycobacterium leprae.

We isolated ancient DNA from skeletal remains obtained from a South German ossuary (approximately 1400-1800 AD) and from a 10th century Hungarian cemetery partially indicating macromorphologic evidence of leprosy. In samples taken of 2 skulls from Germany and of 1 hard palate from Hungary, Mycobacterium leprae-specific fragments of RLEP1 and RLEP3 were amplified using polymerase chain reaction (PCR), thereby confirming their specificity by sequencing. In another case, PCR with primers targeting IS6110 of Mycobacterium tuberculosis gave positive results only for a mandibular specimen. No signal for any mycobacterial DNA was observed in samples from 2 Hungarian foot bones. In ancient material, osseous involvement of M leprae may be detected and distinguished from other mycobacterial infections by specific PCR. In the small bones of leprous hands and feet, not enough M leprae DNA seems to be present for detection. This supports the view that rhinomaxillary leprous alterations result from direct bacterial involvement, while osseous mutilations of hands and feet result from a nervous involvement and/or secondary infections due to small lacerations of the overlying soft tissues.

[Syphilis in Europe in antiquity: the fetus of Costebelle and other new gifts from osteoarchaeology]. / La syphilis en Europe dans l'Antiquité: le foetus de Costebelle et les autres nouvelles données ostéoarcheologiques.

Tomb Nr 1 of the ancient cemetery of Costebelle, attributed to the 4th century AD, contained the skeleton of a pregnant female and that of her foetus in the pelvic cavity. This was aged seven months, was almost complete and showed an exceptional example of bony lesions suggestive of infection. Its etiology suggested the likelihood of early congenital syphilis. This case raises the question of the theory of the importation of venereal disease into Europe, about a 1000 years later, by the crews of Christopher Columbus. The foetus of Costebelle is not an isolated example : other osteo-archaeological findings make a case for the existence of a treponeme (venereal or non venereal) in Europe before 1493.

[Advanced-stage ankylosing spondylitis in a subject in the 8th century]. / Spondylarthrite ankylosante évoluée chez un sujet du VIIIe siècle.

Paleopathological study of an adult male skeleton, coming from a Hungarian archeological site from the 8th century, revealed lesions of the spine (ankylosing vertebral syndesmophytosis, costo-vertebral ankylosis, discal calcifications), of the sacro-iliacs (bilateral ankylosis) and several extraspinal changes (abnormalities of symphysis pubis, enthesopathies). Radiological and CT investigations confirmed the diagnosis of advanced stage ankylosing spondylitis.

IgG isotype-specific auto-antibodies bind preferentially to cross-linked membrane Ig.

Under equilibrium conditions, the affinities of five anti-IgG2a mAb isolated from virus-infected mice were comparable to other high-affinity auto-antibodies. Similar to rheumatoid factors, these anti-IgG2a auto-antibodies bound to aggregated or complexed IgG2a with 50 to 1500-fold higher avidity than their monomeric counterparts. Despite their high functional affinity to IgG2a, flow cytometric analysis revealed no binding or marginal mAb binding to four distinct lines of B cells expressing different densities of membrane-anchored IgG2a. If, however, surface IgG2a was cross-linked by polyclonal light chain-specific antibodies, IgM and IgA mAb binding resulted, and was detected as an increase in mean fluorescence intensity compared with isotype-matched control antibodies. The binding of one IgM mAb to cross-linked IgG2a patches of the cell surface was also visualized by confocal microscopy. Pretreatment of cells with aggregated IgG2a caused increased fluorescence intensity, demonstrating that the IgM and IgA mAb were also able to interact with IgG2a aggregates bound on the B cell surface via Fc gamma RIIB. It also permitted efficient co-ligation of the aggregated B cell receptors (BCR) with Fc gamma RIIB-fixed immune complexes known to deliver a negative signal in B cell activation. Cross-linking of IgG2a complexes bound to Fc gamma RI on macrophages or dendritic cells with antigen-specific BCR and/or T cells via their Fc gamma RIIB may accelerate the physical contact of cells involved in the antigen-specific response.(ABSTRACT TRUNCATED AT 250 WORDS)

Mode of action of SDZ NIM 811, a nonimmunosuppressive cyclosporin A analog with activity against human immunodeficiency virus type 1 (HIV-1): interference with early and late events in HIV-1 replication.

SDZ NIM 811 is a cyclosporin A analog that is completely devoid of immunosuppressive capacity but exhibits potent and selective anti-human immunodeficiency virus type 1 (HIV-1) activity. The mechanism of action of SDZ NIM 811 is clearly different from those of all other anti-HIV agents described so far. In cell-free assays, it is not an inhibitor of reverse transcriptase, protease, integrase, and it does not interfere with Rev or Tat function. SDZ NIM 811 does not down-regulate CD4 or inhibit fusion between infected and uninfected, CD4-expressing cells. p24 production from chronically HIV-infected cells is not impaired either. To elucidate the mode of action of SDZ NIM 811, we performed DNA PCR analysis in HIV-1 IIIB-infected MT4 cells in one cycle of virus replication. The effects of SDZ NIM 811 on the kinetics of viral DNA synthesis, appearance of two-long terminal repeat circles (2-LTR circles), and integration of DNA were studied. SDZ NIM 811 inhibited 2-LTR circle formation in a concentration-dependent manner, which is indicative of nuclear localization of preintegration complexes. Half-maximal inhibition was achieved at 0.17 microgram/ml; this concentration is close to the 50% inhibitory concentrations (0.01 to 0.2 microgram/ml) for viral growth inhibition. As expected, integration of proviral DNA into cellular DNA was also inhibited by SDZ NIM 811. Analysis of the viral particles produced by SDZ NIM 811-treated, chronically infected cells revealed amounts of capsid proteins, reverse transcriptase activity, and viral RNA comparable to those of the untreated control. However, these particles showed a dose-dependent reduction in infectivity (50% inhibitory concentration of 0.028 microgram/ml) which indicates that the assembly process is also impaired by SDZ NIM 811. Gag proteins are postulated to play a role not only in assembly but also in early steps of viral replication, e.g., nuclear localization of the preintegration complex. Recently, it was reported that HIV-1 Gag protein binds to cyclophilin A, the intracellular receptor for cyclosporin A. Interference with Gag-cyclophilin interaction may be the molecular basis for the antiviral activity of cyclosporin A and its analogs.

Modified monoclonal antibody production kinetics kappa/gamma mRNA levels, and metabolic activities in a murine hybridoma selected by continuous Culture.

During long-term continuous culture of the hybridoma cell line 11317, a better-producing subclone (I1317-SF11), giving improved productivity, has been selected. The comparison of the original cell line (I1317-DC) with this subclone revealed that although the growth patterns of both clones were similar, both in continuous and in batch cultures, considerable differences could be seen between the clones with respect to monoclonal antibody (MAB) accumulation, MAB production rate, the levels of mRNA coding for heavy and light chains of IgG, and some metabolic activities. In continuous culture as well as in batch culture, I1317-SF11 showed increased levels of mRNA coding for kappa and gamma chains compared with I1317-DC and/or a modified ratio of the mRNA species when compared to that in I1317-DC. Using pulse experiments, it could be established that the biosynthesis of both chains was augmented in I1317-SF11. Although the kappa and gamma mRNA levels were modified or inversed for I1317-SF11, the cells always synthesized more kappa than gamma chains. The overall increase in the synthetic activity of I1317-SF11 is suggested as one reason for the considerable increase of IgG productivity and product accumulation in continuous culture as well as in repeated batch cultures. Tests concerning metabolic activity revealed that I1317-SF11 had a predominantly glycolytic metabolism independent of growth requirements, whereas for I1317-DC the metabolism became increasingly glycolytic with increased growth. The antibody yield coefficient of I1317-SF11 on glutamine was significantly higher than that of I1317-DC for the continuous culture, whereas the antibody coefficients on glucose were almost similar for both clones under the different culture conditions used. Both antibody coefficients were considerablly influenced by the specific growth rate.All these facts together lead to the conclusion that subclone I1317-SF11 uses more of the energy available, or it was the energy and/or precursors available for the synthesis and production of MAB more efficiently than the thesis and production of MAB more efficiently than the original cell line. Although the levels of mRNA coding for heavy and light chains of IgG were modified, it could be confirmed that the overall regulation of MAB-synthesis and -production occurs post-translationally and that at higher growth rates, more biosynthetic activity is diverted to biomass production.

A new method for the encapsulation of mammalian cells.

A new encapsulation method was developed for the cultivation of mammalian cells. The capsules were produced using a solution of sodium cellulose sulphate (CS)(1.5%) and poly-dimethyl-diallyl-ammonium chloride (PDMDAAC). When CS droplets fell into the precipitation bath consisting of a 2% solution of PDMDAAC, immediately a membrane at the interphase was built up. The influences of varying encapsulation process parameters on capsule characteristics, cell growth, and monoclonal antibody production were tested. This new method showed advantages when compared to other methods mainly due to time simplicity of the whole process.

Application of sensors for the determination of physical and biochemical parameters in the fermentation of animal cells.

Biosensors, sondes, and transducer elements are reviewed with respect to the application for fermentation processes of animal cells. Hereby, the possible connection of these elements to the fermentor is shown. Specially, the (possible) online determinations of the viable and total cell count, of the glucose, the lactate, the pyruvate, and the ammonia concentration in the medium of fermentors are presented. Finally, some sensors for the on-line estimation of the product concentration are described.

Use of a dynamic filtration method for separation of animal cells.

For solving the problem of concentrating and washing of viable cell suspensions a dynamic filter device has been evaluated. Results show that concentrating of cells under sterile conditions can be performed. With a laboratory scale filter (Sulzer-Biodruckfilter BDF-01) a 9-fold concentration was achieved, the filtration speed was about 15 l/h. The viability did not change significantly. Details of experiments are given and advantages are discussed.

Invasive infrared sensor for the determination of the cell number in a continuous fermentation of hybridomas.

A new and simple device for the on-line determination of the total cell count in fermentation vessels is presented. It is based on fibre optic techniques, produces a constant infrared signal and determines the cell count via the scattered light. The linear range between the cell count and the infrared response lies between 1 X 10(5) and more than 2 X 10(6) cells per ml. This sensor shows good correlation between the cell count and the infrared response in fermentations with cell viability of more than 80%. The feeding pumps in these fermentations can be controlled via this sensor to get fermentation with stationary cell counts.

A laboratory fermentor for agarose immobilized hybridomas to produce monoclonal antibodies.

Mass culture of immobilized cells in airlift-fermenters usually ends up with the beads accumulating in the foamy layer on the surface of the reactor fluid or, in stirred tankreactors, with partial destruction of the beads. We tried to use an airlift fermenter vessel for growing cells, immobilized in agarose beads. Instead of using the gas for driving, we mounted a slowly turning marine type impeller within the drought tube. Oxygen was supplied on occasional demands by the original sparger. This set up leads to sufficient operational characteristics of the reactor without accumulation of the beads in the foamy layer and without mechanical destruction. Different productivities of either immobilized cells or cells in free suspension culture are reported.

[Effect of kinetin, 2.4-D and antimetabolites on the changes in amino acid content of withering leaves]. / Die Wirkung von Kinetin, 2,4-DNP und Antimetaboliten auf die Veränderungen im Aminosäurengehalt welkender Pflanzenblätter.

It has been established that in intact plants suffering from scarcity of water as well as in isolated withering leaves the amount of proline increases considerably. The main factor in the isolated leaves is thus the scarcity of water and not the injury due to the defoliation. The proline content of the leaves withering in the dark increases only for a few days and then decreases with the diminution of the carbohydrates. Of the active substances tested only 2,4-DNP inhibited the proline synthesis during the withering. It is very probable that in the course of withering the great amount of proline forms during the oxidation of carbohydrates via α-ketoglutarate. The oxydative phosphorylation is uncoupled by 2,4-DNP. Kinetin, 2,4-D and antimetabolites applied do not inhibit the abnormal increase of proline.