Human mesenchymal stromal cells inhibit platelet activation and aggregation involving CD73-converted adenosine.

BACKGROUND: Mesenchymal stromal cells (MSCs) are promising cell therapy candidates. Clinical application is considered safe. However, minor side effects have included thromboembolism and instant blood-mediated inflammatory reactions suggesting an effect of MSC infusion on hemostasis. Previous studies focusing on plasmatic coagulation as a secondary hemostasis step detected both procoagulatory and anticoagulatory activities of MSCs. We now focus on primary hemostasis and analyzed whether MSCs can promote or inhibit platelet activation. METHODS: Effects of MSCs and MSC supernatant on platelet activation and function were studied using flow cytometry and further platelet function analyses. MSCs from bone marrow (BM), lipoaspirate (LA) and cord blood (CB) were compared to human umbilical vein endothelial cells or HeLa tumor cells as inhibitory or activating cells, respectively. RESULTS: BM-MSCs and LA-MSCs inhibited activation and aggregation of stimulated platelets independent of the agonist used. This inhibitory effect was confirmed in diagnostic point-of-care platelet function analyses in platelet-rich plasma and whole blood. Using inhibitors of the CD39-CD73-adenosine axis, we showed that adenosine produced by CD73 ectonucleotidase activity was largely responsible for the LA-MSC and BM-MSC platelet inhibitory action. With CB-MSCs, batch-dependent responses were obvious, with some batches exerting inhibition and others lacking this effect. CONCLUSIONS: Studies focusing on plasmatic coagulation suggested both procoagulatory and anticoagulatory activities of MSCs. We now show that MSCs can, dependent on their tissue origin, inhibit platelet activation involving adenosine converted from adenosine monophosphate by CD73 ectonucleotidase activity. These data may have strong implications for safety and risk/benefit assessment regarding MSCs from different tissue sources and may help to explain the tissue protective mode of action of MSCs. The adenosinergic pathway emerges as a key mechanism by which MSCs exert hemostatic and immunomodulatory functions.

[Trauma and posttraumatic stress symptoms in patients in German primary care settings]. / Trauma und posttraumatische Belastungssymptome bei Patienten in deutschen Hausarztpraxen.

BACKGROUND: Primary care settings have an important gatekeeping function to detect mental diseases, including trauma and posttraumatic stress disorders. OBJECTIVES: To assess the prevalence of trauma and posttraumatic symptoms in a first sample of northeast German primary care patients and to evaluate the diagnostic sensitivity and specificity of the general practitioners. MATERIAL AND METHODS: Traumatic experiences and posttraumatic stress disorders (PTSD) were assessed with self-rating questionnaires in a sample of N = 400 patients from 3 primary care facilities. Additionally, knowledge and diagnostic accuracy of the general practitioners were evaluated. RESULTS: According to the results of the patient health questionnaire (PHQ-15) data from all patients, the majority of patients questioned showed slight to moderate stress from somatic symptoms. Of the patients with complete data 7 % (n = 25) had a complete PTSD according to the results of the questionnaire, which was also identified in the medical assessment with a sensitivity of 40 %. The stress resulting from posttraumatic symptoms was closely associated with the extent of somatic complaints. CONCLUSION: Patients with a history of trauma and posttraumatic symptoms are prevalent in primary care settings. An early diagnosis by the general practitioner can help patients to receive adequate treatment. Patients with somatoform disorders in particular should be screened for trauma and posttraumatic symptoms.

Role of Chlamydia trachomatis and emerging Chlamydia-related bacteria in ectopic pregnancy in Vietnam.

In this case-control study, we investigated the seroprevalence and molecular evidence of Chlamydia trachomatis and Waddlia chondrophila in ectopic pregnancies (EP) and uneventful control pregnancies in 343 women from Vietnam. Whereas presence of C. trachomatis IgG was strongly associated with EP [adjusted odds ratio (aOR) 5·41, 95% confidence interval (CI) 2·58-11·32], its DNA remained undetected in all tubal lesions. We confirmed an independent association between antibodies against Waddlia and previous miscarriage (aOR 1·87, 95% CI 1·02-3·42). Further investigations are needed to understand the clinical significance of Waddlia's high seroprevalence (25·9% in control pregnancies) in this urban population.

Complete nucleotide sequence of the Oenothera elata plastid chromosome, representing plastome I of the five distinguishable euoenothera plastomes.

We describe the 159,443-bp [corrected] sequence of the plastid chromosome of Oenothera elata (evening primrose). The Oe. elata plastid chromosome represents type I of the five genetically distinguishable basic plastomes found in the subsection Euoenothera. The genus Oenothera provides an ideal system in which to address fundamental questions regarding the functional integration of the compartmentalised genetic system characteristic of the eukaryotic cell. Its highly developed taxonomy and genetics, together with a favourable combination of features in its genetic structure (interspecific fertility, stable heterozygous progeny, biparental transmission of organelles, and the phenomenon of complex heterozygosity), allow facile exchanges of nuclei, plastids and mitochondria, as well as individual chromosome pairs, between species. The resulting hybrids or cybrids are usually viable and fertile, but can display various forms of developmental disturbance.

Sequence variation in the putative replication origins of the five genetically distinct basic Euoenothera plastid chromosomes (plastomes).

Approximately 4,200 nucleotides of the 16S/23S rDNA spacer and the 5' region flanking the rrn operon of the plastid chromosomes representing the five basic, phylogenetically related Euoenothera plastomes were sequenced and compared. The sequences that harbor the putative replication origins are almost identical except for a 785-bp intercistronic segment between the genes for the 16S rRNA and trnI. Differences are mainly caused by insertions/deletions and duplications; the predicted potential for formation of quite extensive secondary structure differs among the plastomes. Unexpected intraplastome variation has also been noted. Furthermore, the sequence-based and published genetically deduced plastome pedigrees differ significantly.

The glutathione S-transferase mu polymorphism as a marker for susceptibility to lung carcinoma.

Glutathione S-transferase (GST) enzymes detoxify carcinogens in tobacco smoke. Interindividual variation in GST function may be related to differences in risk for smoking-related cancer. Leukocytes from 50% of Caucasians lack GST activity toward trans-stilbene oxide (TSO), due to a deletion of the gene for the GST-mu enzyme. Presence of GST-TSO activity in leukocytes has been associated with low risk for lung cancer among cigarette smokers. We sought to determine whether GST activity in lung tissue is determined by the same gene polymorphism and whether it is associated with risk for lung cancer. Subjects were cigarette smokers, identified at the time of lung resection or autopsy in Seattle hospitals. Uninvolved lung tissue was obtained from 35 patients with lung carcinoma and 43 control patients and assayed for GST-mu activity with TSO, for the presence of the GST-mu gene product with an immunological assay, and for the GST-mu gene with Southern blotting. Mailed questionnaires were used to collect information on subjects' smoking histories and exposures which might alter enzyme activity. Interindividual results from the three assays correlated well. Smokers with high GST-TSO enzyme activity present in their lung tissue had a lower risk for lung carcinoma than did smokers with no or low activity (relative risk = 0.30; 95% confidence interval, 0.11-0.79), as did smokers with GST-mu antigen identified in lung tissue versus those with no antigen (relative risk = 0.30; 95% confidence interval, 0.11-0.79). Smokers with both maternal and paternal copies of GST-mu DNA (n = 7) had a lower cancer risk than smokers lacking GST-mu DNA (n = 30; relative risk = 0.35; 95% confidence interval, 0.06-2.10). High GST-mu activity appeared to be associated with a greater decrease in lung cancer risk among 38 heavy cigarette smokers (relative risk = 0.15; 95% confidence interval, 0.03-0.64) than among 38 light smokers (relative risk = 0.61; 95% confidence interval, 0.14-2.60). Presence or absence and number of copies of the GST-mu gene appear to determine activity of the GST-mu enzyme in lung. Smokers with the GST-mu enzyme have approximately one-third of the risk for lung carcinoma of smokers without the enzyme.

Glutathione S-transferase and epoxide hydrolase activity in human leukocytes in relation to risk of lung cancer and other smoking-related cancers.

BACKGROUND: There is considerable interindividual variation in the activity of enzymes which metabolize polycyclic aromatic hydrocarbon constituents of tobacco smoke. Low activity of enzymes which detoxify carcinogenic polycyclic aromatic hydrocarbon metabolites may be associated with increased susceptibility to cancers etiologically related to cigarette smoking. PURPOSE: We conducted a population-based, case-control study to determine whether patients with cancers related to smoking had lower activity of detoxifying isoenzymes of glutathione S-transferase (GST) and epoxide hydrolase (EH) than control subjects. METHODS: Enzyme activities were measured in leukocytes from 113 King County (Washington) residents diagnosed during 1987 with one of three smoking-related cancers (lung, oropharynx/oral cavity, or bladder), 50 King County residents with cancers believed unrelated to smoking (prostate cancer or non-Hodgkin's lymphoma), and 120 persons selected at random from the King County population. Enzyme activity measurements were made for leukocyte cytosolic GST toward transstilbene oxide (TSO), 1-chloro-2,4-dinitrobenzene, and benzo[a]pyrene-4,5-oxide (BaPO), and for microsomal EH toward BaPO. RESULTS: Overall, the distribution of activity levels of GST toward TSO and BaPO did not differ in case patients with smoking-related cancer compared with control subjects. The activities of GST toward 1-chloro-2,4-dinitrobenzene and of EH toward BaPO were somewhat lower on average in case patients with smoking-related cancers than in control subjects, but these differences were well within the limits of chance. Among the heaviest smokers, there were proportionately fewer patients with smoking-related cancers than control subjects with intermediate or high GST activity toward TSO (odds ratio = 0.6), but this difference was also plausibly due to chance (95% confidence interval = 0.3-1.1). CONCLUSIONS: While the findings of this study are compatible with a moderate protective effect of high or intermediate enzyme activity among persons heavily exposed to tobacco, as suggested by an earlier report, the data are by no means conclusive.

Linkage disequilibrium of plasminogen polymorphisms and assignment of the gene to human chromosome 6q26-6q27.

Linkage disequilibrium was observed between newly identified DNA polymorphisms and a previously described protein polymorphism for plasminogen. This finding implies that the two types of polymorphisms describe variation at the same locus. The plasminogen gene was mapped to chromosomal bands 6q26-q27 using somatic-cell hybrids and in situ hybridization. Linkage disequilibrium between protein and DNA polymorphisms has utility in substituting for protein typing in instances where only DNA samples are available, such as from deceased individuals or extinct species. The technique may be useful when cross-hybridizing sequences make the interpretation of Southern blot patterns difficult and may obviate the need for extensive DNA sequencing. In some cases, disequilibrium may provide information useful for determining the appropriate direction for chromosome walks from a marker locus to a target locus.

The association of age, flying time, and aircraft type with hearing loss of aircrew in the Israeli Air Force.

Exposure to aircraft (A/C) noise is considered to be one of the factors which causes permanent hearing threshold shifts among military aircrew. Consequently most studies, dealing with this field of evaluation, fail to focus on the contribution of other elements, especially biological (physiological) ones. We, therefore, decided to evaluate the effect of the subject's age on this process. Audiometric records of 777 aircrew members within the Israeli Air Force were examined. The average age was 27.2 +/- 5.2 years. The individual files were composed of all yearly audiograms from time of recruitment to present, personal information, recorded flight time and type of A/C flown (i.e., rotary-wing and fighter, transport and light fixed-wing aircraft). Hearing loss was studied using two criteria (a. USAF criteria; b. acoustic trauma criteria we devised). The significance of each of the factors was analysed by using uni- and multivariate analysis (Pearson correlation and stepwise multiple regression). A progressive linear correlation between age and hearing loss was exhibited (p less than 0.001). The results suggest that the age of the subject is strongly correlated with values of hearing threshold shift, while A/C type and accumulated time played a minor role.

Linkage disequilibrium and evolutionary relationships of DNA variants (restriction enzyme fragment length polymorphisms) at the serum albumin locus.

Four additional DNA variants (restriction enzyme fragment length polymorphisms) making a total of eight polymorphic sites at the human albumin locus have been identified. These eight sites were found after screening 689 of 20,000 nucleotides by using cDNA probes for albumin with 27 different restriction enzymes. One in 85 nucleotides was therefore potentially polymorphic. The average nucleotide diversity between any two randomly chosen chromosomes was calculated to be 1/500. We observed marked linkage disequilibrium between the eight variants. Only 7 haplotypes among 256 possible combinations were observed in 160 chromosomes from Caucasoids, Blacks, and Asians. Two haplotypes were found in all three human races, indicating that their origin predated human racial divergence. The three rarest haplotypes appear to represent recombinational events between the more common haplotypes. All crossovers occurred in the same general region. Studies of several nonhuman primates indicated that the origin of one haplotype predated the human-African ape divergence. Although it is not possible to rule out maintenance of this tight linkage by selection or fixation, it is suggested that the limited number of haplotypes at the chromosomal site of the albumin gene near the centromere of chromosome 4 may be the result of decreased recombination.

Bone conduction evaluation related to mastoid surgery.

The bone conduction threshold changes of 97 patients (100 ears) who underwent mastoid surgery were determined by comparing the last preoperative audiogram with the 1 year postoperative audiogram. Three types of mastoid surgery were evaluated: radical mastoidectomy, modified radical mastoidectomy, and intact wall atticomastoidectomy. The average three speech frequency preoperative bone conduction threshold was 17.4 dB (S.D. 11.5) in the radical mastoidectomy group, 10.1 dB (S.D. 9.6) in the modified radical mastoidectomy group, and 10.7 dB (S.D. 8) in the intact wall atticomastoidectomy group. The difference between the average three speech frequency preoperative bone conduction threshold of the radical mastoidectomy group differed significantly when compared to the modified radical mastoidectomy or intact wall atticomastoidectomy group. The postoperative average three speech frequency bone conduction threshold did not change significantly following the three surgical procedures evaluated.

Medical air evacuation during the Lebanon War, 1982.

During the Lebanon War, 1982, 85% of the Israel Defence Forces casualties were airlifted from the battle zone by means of the aeromedical evacuation (AME) network of the Air Force. The rescue helicopter was the first point of medical contact for 4.4% of the casualties. A summary is presented of the types of aircraft used in AME and the number of casualties carried in the various flights. A brief description is also given of a new addition to the intensive care capabilities of AME helicopters: the air mobile life support unit used in combat for the first time during this war.

Paraoxon hydrolysis in human serum mediated by a genetically variable arylesterase and albumin.

Gel filtration chromatography resolves human serum paraoxonase into two fractions: (1) a high molecular weight fraction that is completely inhibited by EDTA and coelutes with arylesterase (E.C.3.1.1.2); and (2) a second fraction that is closely associated with albumin, is only partially inhibited by EDTA, and has relatively little arylesterase activity under the assay conditions used. The activity of the high molecular weight fraction is stimulated by NaCl, whereas the albumin associated activity is partially inhibited by NaCl and is not present in serum derived from an analbuminemic individual. Our data suggest that albumin itself, rather than a protein bound to or cofractionating with albumin, mediates paraoxonase activity. The variation in levels of the activity of the nonalbumin, high molecular weight enzyme is responsible for the observed polymorphism of paraoxonase activity in human serum or plasma. An optimal assay of polymorphic paraoxonase activity should be based on activity measurements of the nonalbumin fraction. It is considered likely that only the nonalbumin fraction is responsible for in vivo hydrolysis of paraoxon.

Plasma paraoxonase polymorphism: a new enzyme assay, population, family, biochemical, and linkage studies.

Plasma paraoxonase hydrolyzes paraoxon, the principal metabolite of the insecticide parathione. A genetic polymorphism for enzyme activity has been previously demonstrated. We describe a new assay based on the differential inhibition by EDTA of plasma paraoxonase from persons with the high-activity allele (PX*H) that suggests a trimodality of activity levels in population studies. The gene frequency of the low activity allele (PX*L) in 531 Seattle blood donors of European origin was .7207. Family studies were consistent with codominant autosomal inheritance of two alleles, PX*L (low) and PX*H (high), coding for products with different activity levels. Biochemical measurements of sera from presumed homozygotes for the two different alleles revealed minor physicochemical differences suggestive of a structural difference between the allelic products. No evidence for linkage of the paraoxonase locus with any of 19 polymorphic markers would be detected.

Effect of chronologic age on induction of cystathionine synthase, uroporphyrinogen I synthase, and glucose-6-phosphate dehydrogenase activities in lymphocytes.

The activities of cystathionine synthase [L-serine hydro-lyase (adding homocysteine), EC 4.2.1.22], uroporphyrinogen I synthase [porphobilinogen ammonia-lyase (polymerizing), EC 4.3.1.8], and glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate:NADP+ 1-oxidoreductase, EC 1.1.1.49) have been measured in phytohemagglutinin-stimulated lymphocytes of young and old human subjects. A significant decrease in activity with age was observed for cystathionine synthase and uroporphyrinogen I synthase but not for glucose-6-phosphate dehydrogenase. These changes could not be related to declining phytohemagglutinin response with aging. Age-related decreases in activity of some enzymes may be relevant for an understanding of the biology of aging. False assignment of heterozygosity, and even homozygosity, for certain genetic disorders, such as homocystinuria, may result when low enzyme levels are detected in the lymphocytes of older people.

[Consulting office tissue banks (author's transl)]. / Eine Gewebebank in der HNO-Praxis.

The technicalities of obtaining graft tissues and the methods of preserving homograft ossicles and cartilage are described. The chemical and biological problems of tissue banks are discussed practically in relation to published experiences of others. The techniques for use of homograft ossicles in tympanoplasty surgery are described. The comparative ease of preparation and preservation of homograft ossicles and hyaline cartilage justifies the establishment of tissue banks in the practice of the private otologist.

[A clinical study on allogenetic ossicle-implantation (author's transl)]. / Eine klinische Studie über allogenetische Ossikelimplantationen.

Here we report a first three years clinical experience with allogenetic ossicles in middle ear surgery. The several allogenetic implants in tympanoplasty type III are coordinated to symbolic abbreviations for documentation. The technique and biology of Cialit-preservation is summarized. Findings and results vindicate the application of allogenetic ossicles outside large ENT hospitals too.