Notch-1 regulates NF-kappaB activity in hemopoietic progenitor cells.

We investigated the interaction between two elements critical for differentiation of hemopoietic cells, the Notch-1 receptor and the transcription factor NF-kappaB. These factors were studied in hemopoietic progenitor cells (HPC) using Notch-1 antisense transgenic (Notch-AS-Tg) mice. DNA binding of NF-kappaB as well as its ability to activate transcription was strongly decreased in HPC from Notch-AS-Tg mice. NF-kappaB-driven transcriptional activity was completely restored after transduction of the cells with retroviral constructs containing activated Notch-1 gene. HPC from Notch-AS-Tg mice have decreased levels of several members of the NF-kappaB family, p65, p50, RelB, and c-Rel and this is due to down-regulation of the gene expression. To investigate functional consequences of decreased NF-kappaB activity in transgenic mice, we studied LPS-induced proliferation of B cells and GM-CSF-dependent differentiation of dendritic cells from HPC. These two processes are known to be closely dependent on NF-kappaB. B cells from Notch-AS-Tg mice had almost 3-fold lower response to LPS than B cells isolated from control mice. Differentiation of dendritic cells was significantly affected in Notch-AS-Tg mice. However, it was restored by transduction of activated Notch-1 into HPC. Taken together, these data indicate that in HPC NF-kappaB activity is regulated by Notch-1 via transcriptional control of NF-kappaB.

A single 3' alpha hs1,2 enhancer in the rabbit IgH locus.

Multiple cis-acting elements including the intronic enhancer and the 3'alpha enhancer (3'alphaE) regulate expression of the Ig heavy chain genes during B cell development. A 3'alphaE is composed of DNase I-hypersensitive sites, hs1,2, hs3a,b, and hs4, found 3' of the murine Calpha gene as well as 3' of both human Calpha genes, Calpha1 and Calpha2. Rabbits have 13 Calpha genes, and we tested whether a 3'alphaE is associated with each of these genes. To identify 3'alphaE regions we developed a rabbit hs1,2 probe and used this to search for enhancer homologues of human hs1,2 in a genomic fosmid library. We identified a single hs1,2 fragment 8-kb downstream of Calpha13, the presumed 3'-most Calpha gene. We also identified and partially sequenced a new Calpha gene, Calpha14, located 6 kb upstream of Calpha13. Genomic Southern blot analysis confirmed that the rabbit genome contains only one hs1,2 enhancer region. We tested the enhancer activity of the hs1,2 with the SV40, V(H), and Ialpha promoters using the luciferase reporter gene in transient transfection assays and found that it significantly enhanced the activity of SV40 and V(H) promoters and slightly enhanced an Ialpha promoter. We conclude that the rabbit has a single hs1,2 enhancer that resides at the 3' end of the IgH gene cluster and may constitute one of the cis-elements regulating the expression of IgH genes.

[Problems for discussion on contraception for women after childbirth]. / Diskussionnye voprosy kontratseptsii u zhenshchin posle rodov.

PIP: Questionnaires were administered to 300 postpartum women in 3 delivery wards in Moscow to determine their opinions about contraceptives. Similarly, 100 gynecologists-obstetricians were queried in 2 wards, at 2 counseling sites for women, and at regional seminars of gynecologists-obstetricians. 52% of the physicians recommended the condom to nursing mothers whenever sexual activity was resumed and the IUD from the 3rd and 4th month on. 60% of the doctors thought that women who were not breast feeding could similarly start using the condom and from the 3rd and 4th month either the IUD (82%) or oral contraceptives (36%). 71.1% of the women were under 30 years of age, and were relatively well educated. 68% of them had their first birth, 28.3% had their second, only 3.7% had their third or higher order of births. 35% of the women did not plan the present pregnancy. 81% of the women were using contraception up to the time of the present pregnancy, and the most popular methods were coitus interruptus (36.3%), the rhythm method (35.3%), the condom (28.3%), the IUD (13%), and hormonal methods (10.6%). Although 46.3% of the women preferred the IUD after birth, only 7.3% of them preferred hormonal preparations. In this study 71.5% of the mothers were breast feeding their infants. 13.5% of the women had gestosis in the 2nd half of the pregnancy, 5.8% had anemia, and 21.3% experienced the threat of miscarriage. In 12.9% of the cases there were other pathologies unrelated to pregnancy, and 14% of the births were accomplished by means of cesarean section. Only 12.1% and 13% of the women, respectively, received information about postpartum contraception at the women's counseling sites and delivery wards. 46.3% of the women wanted to use IUDs, but only 7.3% wanted to use hormonal preparations. Only about half of the women expressed their opinion about sterilization: 23% were against it, while 29.7% were for it. The results indicated that the preparation of women for postpartum contraception should already start at the counseling place during pregnancy.^ieng

Internucleosomal chromatin degradation in myeloma and B-hybridoma cell cultures.

The activity of Ca/Mg-dependent endonuclease (CME) is strongly inhibited in myeloma X-63.Ag8.653 and B-hybridoma MLC-1c as compared with mouse splenocytes. Nevertheless, pronounced internucleosomal chromatin degradation occurs in both cell lines during long-term cultivation without passing. In isolated cell nuclei of X-63 the activation of CME, which precedes chromatin fragmentation in vivo and loss of cell viability, is revealed. The time-course of CME activation is opposite to cell proliferation and is not accompanied by alterations in enzyme quantity. The results suggest that cell death of X-63 and MLC-1c occurs via apoptosis, and involves the mechanisms controlling the activation and/or interaction of CME with chromatin.

[The organizational aspects of reducing mortality from abortions]. / Organizatsionnye aspekty snizheniia smertnosti ot abortov.

PIP: Maternal mortality in the USSR in 1988 stood at 43/100,000 population. this figure broke down into mortality from ectopic pregnancy (5%); from induced and self-induced abortion (29.1%); and, from 28 weeks of gestation, mortality from births as well as during the postpartum period (65.9%). Prior to 1991, in 3 years 2020 women died of abortion-related causes, 76.7% of them in connection with non-medical abortions. The original medical documentation of 342 women who died of abortion was analyzed. 69.6% of the women died in connection with non-medical abortion. 72.6% of these women died during the period of 13-27 weeks of gestation. It was especially noteworthy that 58.8% of the women died in cases where their pregnancy was up to 12 weeks of duration and termination of pregnancy could have been performed. Contributory factors were intervention with the purpose of abortion outside of a medical facility, belated provision of medical help, and inadequate equipment at the medical facility. 78/1% of the deaths of non-medical abortions were caused by introduction of catheters or solutions into the uterine cavity and by the opening of the fetal sack through the use of drugs. 32 women carried out the intervention themselves, and 34 were done by strangers, including 18 medical personnel. Sepsis and peritonitis caused 88.6% of the deaths, and hemorrhage caused 6.7%. After induced medical abortions, 44.2% of women died from peritonitis and sepsis, 17.4% from extragenital diseases, and 26.9% from other causes. Only 22.7% of the cases were selected as case studies for medical conferences. Measures regarding the lowering of mortality from abortions have to be directed primarily at the prevention of non-medical or criminal abortions.^ieng

[Abortion as a cause of maternal mortality]. / Abort kak prichina materinskoi smertnosti.

It has been shown that 65.8% of 343 women died in the second trimester and 69.6% after an illegal abortion. Sepsis and peritonitis were the main causes in the general population (73.9%) and in women who died from illegal abortions (88.6%). Defects in medical care were found.

PIP: The medical documents of 342 women who died as a result of abortion (induced, self-induced, and outside of hospitals) were analyzed. 24.2% of the women were under 24 years of age, 51.3% were 25-34 years old, and the rest were 35 or older. 69.9% of the women died after abortion outside of hospitals and the rest after self-induced and induced abortions. 65.8% of the women were pregnant at 13-27 weeks of gestation. The death of 186 of 238 women was due to abortions performed outside of hospitals. Most of these were the result of invasive methods (the introduction of the catheter, solutions, rupture of the amniotic sac). These women subsequently received medical assistance. 93.3% of them were hospitalized with delay, and only 2.6% were admitted in satisfactory condition. In a significant proportion of patients the examination was belated and inadequate. Various laboratory hematological tests that were essential for diagnosis and determination of the severity of the process were not done. The bacteriological tests to select the appropriate antibacterial drug were rarely performed--the tests were done for only 19.8% of women who had died of sepsis and for 14.3% of those who had died of peritonitis. Inadequate care manifested itself in the fact that diagnosis was established for only 46.8% of patients. Sepsis and peritonitis were the cause of death for 73.9% of all women and for 88.6% of those who died of abortion outside the hospital. Hemorrhage was the second most frequent cause, with 42 women dying because of it. Other causes were anaphylactic shock and thromboembolism of the pulmonary arteries. The elimination of such causes of death mandate goal-oriented preventive measures accompanied by diagnosis and therapy and the raising of the knowledge level of the medical personnel about purulent-septic infections. A radical improvement in bacteriological care also must be effected.

[Parturient and puerperal mortality in preterm labor]. / Smertnost' rozhenits i rodil'nits pri prezhdevremennykh rodakh.

The peculiarities of mortality patterns in preterm labour are overviewed, including early and late, induced and spontaneous labour. The revealed drawbacks in the management of future mothers, both in counseling centers and maternity clinics, are discussed.

[Use of monoclonal antibodies in comparative studies of Ca, Mg-dependent endonuclease in cell nuclei]. / Primenenie monoklonal'nykh antitel v sravnitel'nykh issledovaniiakh Ca, Mg-zavisimoi éndonukleazy kletochnykh iader.

Eight hybridoma cell lines derived from fusion between myeloma X-63 and mouse splenocytes were found to secrete monoclonal antibodies against Ca/Mg-dependent endonuclease of human spleen cell nuclei. Two of them, termed N and S, were used in comparative research of enzymes from different organs and species of animals. The data obtained show that N and S antibodies recognize different antigenic determinants of the enzyme molecule. Cross-reactions of antibodies with different antigens having similar antigenic determinants, exist in Ca/Mg-endonuclease of such species as man, mouse, rat and cattle. The evolutionary conservatism of this enzyme is suggested. The data show that the existence of tissue-specific (thymus-specific and spleen-specific) isoforms of Ca/Mg-endonuclease of cell nuclei is possible.

[Dynamics of the endo-DNAase activity of cell nuclei of mouse thymus and spleen lymphocytes during the immune response]. / Dinamika aktivnosti éndo-DNKaz kletochnykh iader limfotsitov timusa i selezenki myshei v protsesse immunnogo otveta.

Endo-DNAse (mostly Ca/Mg-dependent endonuclease) activity was studied in extracts of lymphocyte cellular nuclei from the spleen and thymus of mice upon their immunization with sheep red blood cells. Endo-DNAses were detected by their action on super-stranded DNA pBR 322. It has been established that endo-DNAse activity considerably changes in the course of immune response. The changes start in the early (induction) phase of immune response, are characterized by certain regularities and are distinct in thymus and spleen lymphocytes. It is assumed that endo-DNAses of lymphocyte cellular nuclei are involved in antigen-dependent lymphocyte differentiation.

[Isolation of Ca/Mg-dependent endonuclease of cell nuclei as the basic component in the chromatin nuclease complex of human lymphocytes]. / Vydelenie Ca/Mg-zavisimoi éndonukleazy kletochnykh iader kak osnovnogo komponenta nukleaznofo kompleksa khromatina limfotsitov cheloveka.

Highly purified preparations of Ca/Mg-dependent cell nuclear endonuclease have been obtained from human spleen lymphocytes. The purification was 2000-fold, and the enzyme was a polypeptide with a molecular weight of 57-54 kD. The study of its binding with monoclonal antibody enzyme, produced by various hybridoma strains obtained upon the immunization of mice with non-fractionated nuclear extract has shown that Ca/Mg-dependent endonuclease was the best expressed endonuclease of lymphocyte cellular nuclei. Possible production of some other cell nuclear endonucleases during processing of CH/Mg-dependent endonuclease is suggested.