Leucocytes and intrinsic ROS production may be factors compromising sperm chromatin condensation status.

Considering that the final protection of the DNA against major assaults in terms of chromatin condensation is finalized in the epididymis, it is not known how sperm production of reactive oxygen species (ROS) and inflammatory processes can contribute to protamine deficiency that is predetermined in the testes. Therefore, this study aimed at investigating relationships between poor chromatin condensation, morphology, ROS production, DNA damage and the impact of the presence of leucocytes. In 70 patients, sperm DNA status was determined using TUNEL and chromomycin A(3) (CMA(3)) assays, and ROS-production by means of dihydroethidine. Morphology was evaluated according to strict criteria. The percentage of CMA(3)-positive spermatozoa and leucocyte concentration (r = 0.178, P = 0.0377) as well as percentage of ROS-positive spermatozoa (r = 0.3010; P = 0.012) correlated significantly. Particularly, patients with leucocyte counts >0.5 x 10(6) ml(-1) exhibited higher CMA(3) positivity. No association was found between CMA(3) positivity, TUNEL positivity and sperm morphology. While P- (poor prognosis: 0-4% normal morphology) and G-pattern (good prognosis: 5-14% normal morphology) morphology did not differ regarding chromatin condensation, P-pattern patients had a significantly higher percentage of DNA fragmentation (P = 0.0323). As oxidative stress is associated with disturbed chromatin condensation, results suggest that the idea that under-protamination of sperm DNA will automatically lead to DNA fragmentation might have to be revisited.

Comparative effects of L-tryptophan and 1-methyl-tryptophan on immunoregulation induced by sperm, human pre-implantation embryo and trophoblast supernatants.

PROBLEM: The hypothesis that indoleamine 2,3-dioxygenase (IDO) is necessary to regulate lymphocyte functions at the feto-maternal interface has been postulated, although a possible role of tryptophan (Trp) depletion in the T-cell tolerance during insemination as well as implantation has not been previously investigated. METHOD OF STUDY: Allogeneic phytohaemagglutinin stimulated lymphocytes were supplemented with pre-implantation embryo supernatant (PES), seminal plasma (SP), spermatozoa culture supernatant (SCS), spermatozoa, trophoblast cells, or placenta explant culture supernatants, and analyzed for expression of CD25, CD71, and CD69. Trp-degrading activity was assessed by addition of 1-methyl-Tryptophan or L-Trp. RESULTS: PES, SP, trophoblast, and explant supernatants reduced the expression of CD25 in CD3 lymphocytes. Inhibition of IDO as well as Trp supplementation prevented these effects. CONCLUSIONS: These data suggest that the expression of interleukin-2 (IL-2) receptor in maternal T lymphocytes is normally suppressed by Trp catabolism, and that either abnormal IDO levels or substances influencing IDO activity might lead to non-adequate immune responses on sperm, harm the conceptus or even initiate fetal rejection.

Characterization of a murine model of melioidosis: comparison of different strains of mice.

Melioidosis is an infectious disease caused by the saprophytic gram-negative rod Burkholderia pseudomallei. The aim of this study was to establish and characterize a murine model of melioidosis to provide a basis for further investigations on the pathogenesis of the disease. After intravenous infection with B. pseudomallei, C57BL/6 mice were found to be significantly more resistant than BALB/c mice. There was a marked organotropism of B. pseudomallei for the spleen and liver in both strains of mice, with the highest bacterial load in the spleen. Electron microscopic investigations of the spleen clearly demonstrated intracellular replication within membrane-bound phagosomes. Electron micrographs of the liver provided evidence that B. pseudomallei-containing phagosomes in hepatocytes fuse with lysosomes, leading to degradation of bacteria. In both strains of mice, the course of infection was highly dependent on the infective dose and the bacterial strain used, ranging from death within a few days to death after several weeks. In comparison with BALB/c mice, the bacterial counts in C57BL/6 mice were decreased 12 h after infection, which is suggestive of an innate immune mechanism against B. pseudomallei in this early phase of infection contributing to the lower susceptibility of C57BL/6 mice. BALB/c mice developed a more pronounced lymphopenia, granulocytosis, and splenomegaly at a lower infective dose compared to C57BL/6 mice. Analysis of the antibody response against B. pseudomallei 11 days after infection revealed a significantly higher immunoglobulin G2A (IgG2a)/IgG1 ratio in C57BL/6 mice than in BALB/c mice, indicating that a T helper type 1 immune response is associated with resistance to infection with B. pseudomallei.

[Intracytoplasmic sperm injection--importance and challenge]. / Intrazytoplasmatische Spermieninjektion--Stellenwert und Herausforderung.

The male factor is going to be one of the major problems in the treatment of infertility. In addition to the standard methods of assisted reproduction new microtechniques of assisted fertilization have been developed to increase success rate in cases of male factor infertility. Of these methods especially intracytoplasmatic sperm injection (ICSI) has been described as beneficial in which one spermatozoa is injected directly into oolemma of an oocyte. ICSI can be carried out with ejaculated spermatozoa, with epididymal spermatozoa and with spermatozoa isolated from a testicular biopsy. In all cases high fertilization and pregnancy rates comparing with standard in-vitro fertilization were achieved. It may be recommended that the couples undergo prenatal diagnosis and participate in a prospective follow-up study of children born after ICSI. In management of ICSI all specialists in gynecology, embryology, andrology, medical genetics and urology should go together.

HLA and thyroid carcinoma.

HLA typing was performed in 40 thyroid carcinoma patients. No association was found between antigens of the HLA system and papillary, follicular or medullary thyroid carcinomas. Literature reports on different HLA associations are probably due more to the small number of cases in the studies involved than to ethnic or geographic differences.

[Fatal course of immune hemolytic anemia following administration of diclofenac]. / Eine tödlich verlaufene immunhämolytische Anämie nach Applikation von Diclofenac.

We report of a case of a 56 years old female patient who died of severe haemolytic anaemia induced by an injection of a single dose of diclofenac. We suppose that an immune complex reaction caused the haemolytic crisis.

[Treatment of long-term sterility by intra-tubal gamete transfer]. / Die Behandlung langjähriger Sterilität durch intratubaren Gametentransfer (GIFT).

It is reported about a first successful program of gamete intra fallopian transfer (GIFT) as a supplement to intrauterine insemination and to in vitro fertilization (IVF). In 29 patients after HMG stimulation of the ovaries and laparoscopic aspiration of follicles recovered oocytes have been introduced together with prepared sperm via a special catheter directly into the ampulla of the oviduct. In 21 patients with different causes of infertility optimal stimulation conditions could be achieved. A pregnancy rate of 41.4% is the result of 29 GIFT events and 3 of them resulted in spontaneous term deliveries (10.3%).

Hepatitis safety of a new prothrombin complex preparation, sterilized according to the method of LoGrippo. Results of a prospective clinical trial.

In a controlled, prospective clinical trial with almost 300 patients undergoing cardiovascular surgery, the hepatitis risk was examined of a new prothrombin complex (PPSB) preparation produced from pooled plasma, which was cold sterilized according to the method of LoGrippo with beta-propiolactone and UV-light. Of 268 evaluable patients, 102 belonged to a group receiving PPSB and 166 belonged to a control group without PPSB. 25 batches of PPSB in dosages of up to 4,500 units per patient, in total, were used. The mean requirement for transfusion of blood or plasma was 10 units per patient in the group receiving PPSB and slightly more than 2 units per patient in the control group. In both of the groups, 3 hepatitis infections occurred (5 cases of non-A, non-B, 1 case of type B), corresponding to hepatitis incidences of 2.9 and 1.8%, respectively. The difference in hepatitis incidence between both groups was statistically not significant. This demonstrates, on the one hand, the hepatitis safety of the sterilized PPSB and, on the other, that multiple transfusion is no longer associated with a high risk of hepatitis when the guidelines of modern donor screening are followed.

Effects of a fixed combination of low dose nifedipine and acebutolol on essential hypertension: comparison with standard dose acebutolol.

114 patients from four clinics participated in a double blind study designed to assess the efficacy of a nifedipine-acebutolol fixed combination -10 mg + 100 mg - as compared with acebutolol -200 mg- in essential hypertension. During the ten week study the mean blood pressure readings (s.d.) 1-3 h after treatment decreased from 179.2/104.8 (10.2/6.2) to 150.3/87.7 (9.8/7.7) in the combination group and from 181.7/106.5 (14.4/7.0) to 150.4/89.0 (15.0/10.4) in the acebutolol group. The mean systolic and diastolic blood pressures were also decreased after exertion (load) and 24 hours after treatment at the end of the 6th week of the study. A doubling of the dose from week 7 to 10 did not change these figures. These results reveal the possibility of treating essential hypertension with a low dose of beta-adrenergic blocking agents in combination with 10 mg nifedipine. Both drugs were well tolerated. 3 patients (5%) in the combination group and 3 patients in the acebutolol group were withdrawn from the study because of headache and dizziness.

Prevention of fetal growth retardation by buffy-coat transfusions--a case report.

A case is described with two previous unsuccessful pregnancies (one missed abortion and one severe fetal intrauterine growth retardation ending in fetal death). Examination showed that the patient and her husband each share one antigen of the HLA-A and -B series. In spite of two previous pregnancies the patient had no lymphocytotoxic antibodies. The patient then received 5 buffy-coat-pool transfusions and the sera reacted with an increasing number of panel cells (Fig. 1). A mature healthy infant was born at term. We consider that the cytotoxic antibodies achieved, took over a protective function for the later undisturbed progress of the pregnancy. This may be a possible way of preventing intrauterine growth retardation caused by immunologic factors.

[Risk of hepatitis from conventional pooled PPSB preparations]. / Hepatitisrisiko von konventionellen PPSB-Poolpräparaten.

The risk of hepatitis from prothrombin complex (pooled human clotting preparation) was retrospectively analysed. Of 39 patients who had undergone cardiac surgery, 22 (56%) developed hepatitis, while in those had not received pooled preparations the rate was only 5%. Although the proportion of multiple transfusions was significantly higher among the recipients of clotting preparations, it was found that the decisive hepatitis-inducing factor was the pooled preparation, not the transfusion blood. Contrary to earlier results, all cases were of the non-A, non-B type. The frequency of carriers of the causative virus is apparently not different from that with B virus. Thus both virus types must have occurred at similar frequency in earlier pooled clotting preparations. Since, furthermore, there seems to be no difference in their infectivity and their penetration in the population at large is likely to be similarly low, unrecognised double-infections in recipients of pooled clotting preparations were probably frequent before the introduction of recent methods of demonstrating hepatitis B.

[Hospital-acquired serum hepatitis (author's transl)]. / Krankenhaus-Hepatitis.

During a prospective study of blood transfusions at the Cardiac Surgery Unit of the University Surgical Clinic, Free University, Berlin, a patient developed hepatitis B although he had not received any blood transfusion. Soon after, two further patients developed B-hepatitis and a third had HBs-antigenaemia. A virus marker confirmed the infectious pathways. None of the three infected patients had received foreign blood or large-pool clotting preparations. These special circumstances made it possible to diagnose hospital-acquired serum hepatitis. Otherwise all three cases would probably have been considered as caused by transfusion of foreign blood or blood products. It is, therefore, recommended that in similar cases one should speak of hospital-acquired hepatitis and of transfusion hepatitis only when this can be proven.

Hepatitis after cardiosurgery. Frequency and causes. Results of a prospective study with use of autologous blood.

In a prospective study on the causes and frequency of hepatitis after operations on the open heart, two homogeneous groups of patients were formed. In one of them, the amount of homologous blood was reduced to half by means of preoperative donations of autologous blood, the total blood requirement remaining the same. Hepatitis occurred postoperatively in 10% of the cases. The factors which gave rise to the hepatitis were large-pool clotting preparations, transfusion with homologous blood and cross-infection in the hospital. Even after exclusion of other risk factors, the frequency of hepatitis among the recipients of coagulation preparations was around 60%. The frequency of the cases of hepatitis due to foreign blood was about 5 per 100 patients or about 1 per 100 units of blood. By use of autologous blood collected preoperatively, the risk due to transfusion could be lowered by 50%. In polytransfusion, a disproportionately high rise in the risk of hepatitis was observed. It is probably to be ascribed to unknown factors in the hospital environment ("nosocomial hepatitis").

Deficiency of antibody formation to HBsAg in the pathogenesis of chronic hepatitis and cirrhosis?

UNLABELLED: Sera of 832 healthy persons and patients suffering from chronic inflammatory liver disease were investigated by radioimmunoassay for HBsAg and anti-HBs. Diagnosis in patients was secured by biopsy. The persons were divided into: 1. Healthy persons: n = 478 blood donors, hospital especially exposed to HBV, patients with healed hepatitis; 2. PATIENTS: n = 354 acute hepatitis, chronic persistent and aggressive hepatitis, post-hepatitic, cryptogenic and alcoholic cirrhosis. The results demonstrate considerable accumulation of HBsAg in chronic liver disease (72% in CAH, 66% in posthepatic liver cirrhosis) whereas anti-HBs was more frequently observed in healthy persons (38% in hospital staff, 49% in healed hepatitis). Furthermore, HBsAg and anti-HBs were frequently observed simultaneously in chronic hepatitis and cirrhosis (23% in CAH). A strong shift in the relation of antigen to antibody to the disadvantage of antibody in the examined collectives of chronic hepatitis and cirrhosis is evident. Chronic inflammatory HBsAg positive liver disease should therefore be regarded as chronic virus infection. We suppose an absolute or relative deficiency of antibody to HBsAg is probably an important factor for the development of chronicity of hepatitis B.

[Antibody screening of commercially available immunoglobulins. Erythrocyte-, HLA- and autoantibodies (author's transl)]. / Antikörperscreening an kommerziellen Immunglobulinpräparaten. Erythrozyten-, HLA- und Autoantikörper.

A great number of commercially available immunoglobulins was screened for erythrocyte-, HLA- and autoantibodies. Isoagglutinin titers were in general low, hemolysins were not detected. No preparation was found to contain detectable amounts of autoantibodies versus nuclei, mitochondria, smooth muscle or parietal cells. Yet 7 out of 22 immunoglobulin charges contained irregular red cell antibodies, aside of polyspecific albumin reactive types in 5 cases a distinct anti-D. Only the preparation examined in 1975 contained polyspecific HLA-antibodies. 1977/78 all preparations for intravenous use were free from undesired antibodies, while those for intramuscular application were contaminated with irregular red cell antibodies 1975 as well as 1978. Because of their low strength these antibodies imply no clinical risk. Nonetheless an improvement of the official rules for the production and control of immunoglobulin preparations is inevitable to avoid damage caused by exceptional charges with extremely strong antibodies of this type.

HLA-antigens in systemic lupus erythematosus (SLE).

Histocompatibility antigens were determined in 27 patients with SLE and 283 normal blood donors. HLA-B8 was significantly increased in patients with SLE (59.3%) as compared to the controls (20.5%, Pc less than 0.005). The relative risk for HLA-B8 was 5.64. No correlation could be demonstrated to specific organ involvement, immunopathological data, age of onset or prognosis.

A refined map of the hisG gene of Salmonella typhimurium.

The hisG gene is the most operator-proximal structural gene of the histidine operon; it encodes the feedback-inhibitable first enzyme of the biosynthetic pathway. Previously, hisG mutants were mapped into seven intervals defined by the availble deletion mutations having endpoints in the hisG gene. The map has been refined using over 60 new deletion mutants. The new map divides the gene into 40 deletion intervals, which average approximately 30 base pairs in length. The map has been used to analyze the distribution of insertion sites for the transposable element Tn10 and has permitted conclusions on the diistribution of duplication endpoints. The map promises to be useful in analysis of his regulation and, more particularly, in the determination of the possible role of the hisG enzyme in this mechanism.