Endotoxin removal from whole blood by a novel adsorption resin: efficiency and hemocompatibility.

The structural component of Gram- bacteria, endotoxin (ET), induces the release of endogenous mediators of sepsis. Attempts to remove these downstream molecules in vivo, have not improved survival. However, extracorporeal strategies such as continuous renal replacement therapy or therapeutic plasmapheresis have shown benefit. We are presenting an affinity-based extracorporeal technology for the removal of ET from whole blood. The small-scale device contains an adsorbent that removed 75% of ET present in whole blood. This affinity resin displayed good hemocompatibility regarding the coagulation pathway. Minimal platelet, neutrophil and complement activation were observed. There was also no evidence of consumption of coagulation factors or cell loss. In as much as ET participates in both the inflammatory and coagulation abnormalities in sepsis, this method represents an efficient and hemocompatible way to remove ET from whole blood, which, in an extracorporeal setting, may improve the outcome of sepsis.

HLA alleles and susceptibility to dermatological disorders associated with Helicobacter pylori infection: a significant association to HLA-Cw*06.

The occurrence rate of HLA class I and class II alleles was established in 24 patients suffering from dermatological disorders associated with the Helicobacter pylori infection. The increased frequency of HLA-C*0602, 4 was found to be 0.1875 compared to 0.0733 in the control group (odds ratio: 2.913; two-sided P value: P = 0.0251). Our data suggest that the HLA-Cw6 molecule play a role in the susceptibility to the Helicobacter pylori infection.

[Trends in occupational skin diseases in the Slovak Republic]. / Vývojové trendy kozných chorôb z povolania v Slovenskej republike.

The authors present the onsets of occupational dermatoses abroad and in the Slovak Republic, organizational measures, the so-called consulting days of commissions which co-operate with medical centres of occupational hygiene. They analyse the principles of diagnosis, relevance of epicutaneous tests, criteria of professionality, claims service, judgement and compensations of admitted claims. They analyse the developmental trends of claimed occupational skin diseases and skin infections which occurred within 1973-1998 according to their individual inducers divided into 20 classes, as well as to number of claims. They review the order of inducers according to their significance: oil products, plastic materials, rubber and rubber chemicals. The 4th class includes "other chemicals" which are to be divided according to their individual chemical composition. The proportion of occupational skin diseases in relation to all occupational diseases has gradually decreased from 51.7% in 1973 to 16.3% in 1998, in absolute figures from 382 cases in 1973 decreased to 60 cases in 1998. The authors analyse the reasons of this decrease. (Tab. 4, Ref. 39.)

Occurrence rates of HLA-DRB1, HLA-DQB1, and HLA-DPB1 alleles in patients suffering from vitiligo.

By investigating a group of 39 unrelated adults suffering from vitiligo it was found that alleles HLA-DRB1*0701, -DQB1*0201, and -DPB1*1601 differed in their frequencies in comparison to those observed in the healthy population. The allele HLA-DRB1*0701 was found in 26.5% of patients compared to 14.2% in the healthy group (p < 0,01, RR = 2.17). The allele HLA-DQB1*0201 was present in 33.8% of patients compared to 21.2% (p < 0,025, RR = 1.89). The allele HLA-DPB1*1601 was found in 6.41% of patients compared to 2.05% in the healthy group (p < 0.05, RR = 3.3). No other significant deviations in the frequencies of investigated alleles were observed.

Long-term trends of the incidence of syphilis and gonorrhoea in the Slovak Republic during 1947-1994, with special regard to young age groups.

During the Second World War and after the War venereal diseases (Syphilis and Gonorrhoea) were wide-spread. By a single census in October 1947 were notified 2960 syphilitic patients. In the years 1949-1951 intensive and effective preventive measures were introduced as compulsatory notification system of venereal diseases, sexual contact-tracing investigations, correct and rapid diagnosis of venereal diseases, rapid and efficient therapy of patients with syphilis, followup (post-treatment observations) for all patients, routine serological tests for syphilis, in expectant mothers at ante-natal clinics, periodical screening of high-risk groups, patients in each calendar year at their hospitalization, to taking up a post and a special drive against syphilis ("PN-action"). The results of these measures are manifested by substantial fall in the number of cases of syphilis and gonorrhoea. In the years 1948-1955 18,151 patients suffering from syphilis were notified. The changes of the incidence of syphilis and gonorrhoea are illustrated by statistical data (Tab. 1-9). Syphilis congenita occurs sporadicly, there is a substantial fall in the number of syphilitic pregnant. After 1989 there has been a greater group mobility and a substantial rise in active and passive tourisms and occupational travel activity, especially abroad. Changes of the sexual behaviour were expressed, especially a greater tolerance whether heterosexual or homosexual, sexual freedom, sexual promiscuity and especially prostitution.

Vitiligo is associated with HLA-A2 and HLA-Dw7 in the Slovak populations.

By investigating a group of 67 unrelated adult persons suffering from vitiligo it was found that antigens HLA-A2 and HLA-Dw7 differed in their frequencies in comparison to those observed in the healthy population. The antigen HLA-A2 was found in 76.12% of patients compared to 43.95% in the control group (chi 2 = 25.61, P < 0.005, RR = 4.07). The antigen HLA-Dw7 was present in 56.71% of patients compared to 15.8% in the healthy population (chi 2 = 26.55, P < 0.0001, RR = 6.98). No other significant deviations in the frequencies of investigated antigens were observed.

[Adverse effects and side-effects of antimycotic agents]. / Neziaduce a vedl'ajsie úcinky antimykotík.

The study lays emphasis on the significance and actuality of infections, especially in coincidence with immunodeficient states. Candidosis, mycotic diseases caused by moulds, systemic mycoses, and onychomycoses represent the major problems of modern mycology. They require usually a long-term therapy by means of highly active antimycotics. Consequently, an entire series of undesirable side-effects supervene. External applications result most frequently in contact eczemas and therefore often require preventive patch tests. Oral administration results in general hepatic, renal, hematologic and other impairments which require the monitoring of the latter parameters during therapy. Finally a considerable significance is ascribed to interaction with other simultaneously applied drugs with the antimycotics. (Tab. 1, Ref. 83.)

[Vitiligo--an enigmatic disease. II. Therapy of vitiligo]. / Vitiligo--záhadná choroba. II. Liecba vitiliga.

The work gives a review of therapeutic methods of vitiligo. PUVA therapy, application of cortisonoids, khellin (including UVA), phenylalanine (including UVA), melagenina, and laser therapy represent non-surgical methods. Surgical methods include application of fluorouracil following dermabrasion, of autologous epidermal grafting transplantation of in vitro cultured epidermis, in vitro cultured melanocytes and micropigmentation by pigment. Auxiliary methods are represented by administration of carotene, using sunscreens and if necessary cosmetic treatment. Entire depigmentation is done in accordance with the extent of vitiligo, unsuccessful repigmentation and the age of the patient. The conclusion points out psychosocial aspects and the necessity of mutual cooperation of both, physician and patient. (Ref. 53.)

In vitro effects of hematopoietic growth factors on the proliferation, endoreplication, and maturation of human megakaryocytes.

A liquid culture technique was used to study regulation of human megakaryocytopoiesis in vitro. Low-density cells from adult bone marrow were cultured in the presence of normal plasma, plasma from patients with aplastic marrows (AP), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and interleukin-3 (IL-3). Megakaryocytes (MK) were studied at day 10 of culture by a two-color staining technique using a pool of monoclonal antibodies for their identification and propidium iodide to label DNA. Their ploidy distribution was analyzed by flow cytometry. In some experiments cytoplasmic maturation was also studied by ultrastructural techniques. Normal plasma provides a low number of MK with a ploidy distribution including 8 N and 16 N MK. AP promoted in a dose-dependent manner proliferation of MK and some batches favored endoreplication. This effect was clearly demonstrated when ploidy distribution was compared between normal plasma and AP on parallel marrow cultures. However, ploidy distribution was shifted toward low values compared with uncultured MK. rhGM-CSF had no significant effect on these two parameters. In contrast, rhIL-3 from 0.1 U/mL to 100 U/mL had a proliferative effect but was unable to induce endoreplication. Furthermore, when associated with AP it totally abrogated the effect of AP on endoreplication because in most experiments more than 90% of MK were 2 N and 4 N. This effect was also observed when rhIL-3 was added after 7 days of culture (when it has little proliferative effects). Studies of the maturation of MK grown with rhIL-3 indicate that the majority were small mature cells synthesizing alpha-granules and demarcation membranes. The effect of AP on MK proliferation and endoreplication was not related to IL-6 because its IL-6 content was identical to that of normal plasma and its neutralization did not modify these parameters. In conclusion, this study indicates that liquid culture technique in association with flow cytometry could be a powerful tool in identifying the humoral regulators of human megakaryocytopoiesis.

Developmental changes in human megakaryocyte ploidy.

Megakaryocytes (MK) obtained from the differentiation of MK colony-forming units (CFU-MK) were grown from fetal liver, cord blood, and adult marrow in liquid culture containing aplastic plasma. Ploidy distribution was studied by a double-staining technique and flow cytometry and MK maturation by ultrastructural techniques. Cultured MK from fetuses and neonates were small sized (about 10 microns) in comparison to adult MK. They were mature cells that contained large membrane complexes as previously found in vivo. Only 2N and 4N MK were usually present in 8- to 10-week-old fetus cultures; 8N MK were detected at 20 weeks of gestation and in neonates. Higher ploidy classes were present in culture from adults but with a much lower frequency than in marrow. Therefore, a progressive shift to higher ploidy and an increase in MK size were observed simultaneously during development. Interleukin 3 (IL-3) increased MK proliferation as in adults but abrogated MK ploidization of 20-week-old fetus culture. The present results suggest that the changes occurring during ontogenesis are related to intrinsic MK modifications because no inhibitor of MK ploidization could be detected in fetal cultures.

[Propolis allergy]. / Zur Frage der Propolisallergie.

The incidence of propolis contact sensitivity was 1.2-3.3% among 7483 hospital patients during the period 1981-1987, while the prevalence among 1,558 healthy volunteers was 0.64%. A group of 26 patients with contact allergy to propolis was challenged with identified substances isolated from propolis. The mixture of 3-methyl-2-butenylester (3M2B) and 3-methyl-3-butenylester (3M3B) from caffeic acid caused a positive reaction in 7 patients, and in 5 of 9 patients there was a positive reaction to the mixture of 3M2B and 3M3B from diacetyl-caffeic acid. Among the 19 patients challenged with the flavonoid group, 3 had positive reactions, and individual derivatives of cinnamic acid caused positive reactions in 3 and 4 patients each. When five components taken from poplar buds were tested, the one that provoked the largest number of positive reactions was the methanol component (15 of 19 patients). The results show that propolis contact allergy is not caused by one main allergen, but by several allergens varying in chemical composition; the presence of these in propolis depends on the nature of the source plant and the place and time of collection by the bees.

Immunogold probing of platelet factor 4 in different ploidy classes of rat megakaryocytes sorted by flow cytometry.

Different ploidy classes of rat megakaryocytes were sorted by flow cytometry from highly purified perfusion-fixed megakaryocyte cell suspensions prepared by sequential centrifugal elutriation and Percoll gradient centrifugation. Sorted cell populations were studied for the localization of platelet factor 4 (PF-4) probed with the monoclonal antibody 2E7 in order to clarify the relevance of PF-4 localization to the cytoplasmic and nuclear development of megakaryocytes. The relative numbers of labeled alpha granules and labeled alpha granule-related small vesicular structures (AGR-SVS) were quantitated using the gold-labeled antibody detection method and correlated with DNA content and cytoplasmic maturation in individual megakaryocytes. We determined that the stage of cytoplasmic maturation exerted a significant effect on the proportion of labeled alpha granules and labeled AGR-SVS. A significant interaction effect of stage and ploidy class resulted in the stage effect on proportion of labeled alpha granules being significant only in two of the three ploidy classes. The least mature cells present within each ploidy group exhibited PF-4 labeling mostly in SVS that were not related to alpha granules. During subsequent cytoplasmic maturation, more of the labeled SVS were seen related to alpha granules, with more of the mature alpha granules themselves becoming labeled. Polyploidization also affected the proportion of labeled AGR-SVS. Our data suggest that SVS play a role in the intramegakaryocytic transport of PF-4 into alpha granules. These data provide evidence of the complexity of megakaryocytic differentiation involving both cytoplasmic maturation and nuclear endoreduplication as reflected in PF-4 expression.

Regulation of human megakaryocytopoiesis: analysis of proliferation, ploidy and maturation in liquid cultures.

A liquid culture technique associated with either double staining and flow cytometry or electron microscopy was used to study human megakaryocytopoiesis. During development from the embryo to the adult, a progressive increase in ploidy classes associated with an enhancement of megakaryocyte (meg) size was observed. Granulocyte-macrophage colony-stimulating factor had no effects on adult marrow cultures. In contrast, interleukin (IL) 3 induced a marked proliferation, but was unable to promote polyploidization. Furthermore, it abrogated the effects on endomitosis of aplastic plasma (AP). This negative effect on polyploidization of IL-3 could be partially dissociated from its effects on proliferation by a delayed addition in culture. AP acted on both proliferation and endoreplication, which was not due to the main hematopoietic growth factors, including IL-6. A synthesis of IL-6 was detected by in situ hybridization in cultured cells including megs which also express receptors for IL-6. These results suggest that terminal meg differentiation may be regulated by an autocrine IL-6 loop, and that megakaryocytopoiesis may be independently regulated at early and late stages of differentiation.