Zootechnical and genetic aspects of a prolific merino program.

In a Prolific Merino nucleus herd of 200 ewes the ovulation rate (OR) test results obtained in 169 animals between 1988 and 1993 were compared with those of 113 ewes from the same herd in 1996. Whereas earlier the ratio of individuals showing an OR > or = 4 was only 32%, that of the group checked in 1996 was 59%. This increase could be attributed to 40 ewes, both of whose parents had proven to be homozygous carriers of the prolific gene. To develop the Prolific Merino breed, 21 Booroola Merino rams were imported from New Zealand, and mostly their frozen semen was used. Of these rams, one was not a prolific gene carrier, 8 were homozygous carriers, 10 were heterozygous carriers and two had not been identified yet. Of the 36 home-bred rams, 9 proved to be homozygous by parents, 11 heterozygous, 8 homozygous, one proved to be a non-carrier, and 7 rams and their frozen semen were to be progeny tested. Six thousand doses of frozen semen from a total of 33 animals (16 imported rams and their 17 home-bred offspring) are stored in plastic straws. Sixty-three % of this is semen reserve from rams of the FecBFecB genotype, belonging to 10 ram lines. The remaining 37% is gene reserve intended for creating homozygous ram lines. Only one ram (no. 3244) was bought for the nucleus herd, the other ram lines were introduced into the herd by assortative mating, using intrauterine insemination. The average conception rate found after 472 intrauterine inseminations was 53% with large (occasionally 10-100%) individual ram differences.

Astrocyte-targeted expression of IFN-alpha1 protects mice from acute ocular herpes simplex virus type 1 infection.

Type I IFNs (i.e., IFN-alpha and IFN-beta) play a key role in the host's innate defense against viral pathogens. To examine the biologic relevance of IFN-alpha to a viral pathogen within the confines of the nervous system, IFN-alpha1 transgenic mice whose transgene is under the control of the glial fibrillary acidic protein promoter (GFAP-IFN-alpha, astrocyte specific) were examined for resistance to an ocular herpes simplex virus type 1 (HSV-1) infection. GFAP-IFN-alpha mice expressed significantly higher levels of IFN-alphabeta (533 U) in the trigeminal ganglion compared with nontransgenic mice (70 U) 72 h postinfection that corresponded with a significant reduction in the mRNA expression of the HSV-1 immediate early gene infected cell polypeptide 27 and late gene VP16, as well as the chemokines monocyte-chemoattractant protein-1 and cytokine response gene-2 in the eye and trigeminal ganglion. Six days postinfection, the viral load and the expression of infected cell polypeptide 27, CD8, RANTES, IFN-gamma, and IFN-alpha mRNA levels were reduced in the trigeminal ganglion of GFAP-IFN-alpha mice compared with the wild-type mice. Following the establishment of HSV-1 latency (i.e., 30 days postinfection), only one of nine (11%) GFAP-IFN-alpha mice was found to be latent compared with seven of eight (88%) of the wild-type mice, as determined by the expression of the latency-associated transcript RNAs. Likewise, only three of nine GFAP-IFN-alpha mice screened showed seroconversion by day 30 postinfection compared with nine of ten wild-type mice screened. Collectively, the results show that the IFN-alpha1 transgenic mice are less susceptible to acute HSV-1 infection and the establishment of viral latency.

The persistent elevated cytokine mRNA levels in trigeminal ganglia of mice latently infected with HSV-1 are not due to the presence of latency associated transcript (LAT) RNAs.

Trigeminal ganglia (TG) from mice latently infected with wild type HSV-1 contain detectable levels of cytokine transcripts that are not present in TG from uninfected mice. This suggests that during HSV-1 neuronal latency, the immune system is stimulated by the production of one or more viral proteins. Since the LAT (latency associated transcript) gene is essential for wild type levels of spontaneous reactivation and is the only highly active viral gene during latency, the stimulation of cytokines may indicate the presence of a LAT encoded latency protein. We therefore compared the cytokine transcript profiles in the TG of mice latently infected with wild type and LAT negative viruses. Mice were latently infected with either: (1) the LAT null mutant dLAT2903; (2) its marker rescued virus dLAT2903R; or (3) the parental wild type HSV-1 strain McKrae. As expected, reactivation following explant cultivation of TG from latently infected mice was significantly decreased with dLAT2903 (P < 0.05)(40 +/- 8%, n = 24) compared with dLAT2903R (85 +/- 7.6%, n = 36) or the parental virus (70 +/- 10.0%, n = 36). The relative levels of various cytokines was determined by RT-PCR analysis of TG extracts. None of the cytokine transcripts detected in mice latently infected with the wild type or marker rescued viruses were missing or decreased in mice latently infected with the LAT null mutant 30 or 60 days post infection. There were also no differences in the HSV-1 antibody titers induced by the LAT negative virus compared to the LAT positive viruses. Thus, although LAT facilitated reactivation of HSV-1 from explanted mouse TG, expression of LAT during latency did not appear to be involved in persistent cytokine expression in TG. This suggests that during latency, HSV-1 does not produce a highly antigenic abundant LAT encoded protein.

Role of the hypothalamic pituitary adrenal axis and IL-6 in stress-induced reactivation of latent herpes simplex virus type 1.

Hyperthermic stress induces reactivation of herpes simplex virus type 1 (HSV-1) in latently infected mice and also stimulates corticosterone release from the adrenals via activation of the hypothalamic pituitary adrenal axis. In the present study, we tested the hypothesis that stress-induced elevation of corticosterone potentiates HSV-1 reactivation in latently infected mice. Because of the putative role of IL-6 in facilitating HSV-1 reactivation in mice, the effect of hyperthermic stress and cyanoketone treatment on IL-6 expression in the trigeminal ganglion was also measured. Preadministration of cyanoketone, a glucocorticoid synthesis inhibitor, blocked the stress-induced elevation of corticosterone in a dose-dependent manner. Furthermore, inhibition of corticosterone synthesis was correlated with reduced levels of HSV-1 reactivation in latently infected mice. Hyperthermic stress elicited a transient rise in IL-6 mRNA levels in the trigeminal ganglion, but not other cytokine transcripts investigated. In addition, there was a significant reduction in MAC-3+, CD8+, and DX5+ (NK cell marker) cells in the trigeminal ganglion of latent HSV-1-infected mice 24 h after stress. Cyanoketone blocked the stress-induced rise in IL-6 mRNA and protein expression in the trigeminal ganglion latently infected with HSV-1. Collectively, the results indicate that the activation of the hypothalamic pituitary adrenal axis plays an important role in stimulating IL-6 expression and HSV-1 reactivation in the trigeminal ganglion following hyperthermic stress of mice.

Immunization with HSV-1 antigen rapidly protects against HSV-1-induced encephalitis and is IFN-gamma independent.

Herpes simplex virus type 1 (HSV-1) infection of mice frequently culminates in fatal encephalitis. Intraperitoneal administration of heat-inactivated HSV-1 0-5 days before infection (active immunization) protected mice from encephalitis. In addition, active immunization 2-5 days before ocular infection with HSV-1 reduced the frequency of establishment of latent HSV-1 infection in the trigeminal ganglion (TG). However, intraperitoneal administration of heat-inactivated HSV-1 did not induce interferon (IFN) production in the peritoneum or serum, as determined by bioassay and ELISA. Intraperitoneal administration of heat-attenuated HSV-1 elicited IFN-gamma but not type I IFN production in the peritoneum. The production of IFN-gamma correlated with the infiltration of CD4 and CD8 cells in the peritoneum as determined by RT-PCR. In addition, there was a significant increase in interleukin (IL)-12 p40, IL-12p35, IL-6, IL-10, and IFN-gamma mRNA in peritoneal cells, as determined by RT-PCR following immunization with heat-attenuated HSV-1, which was not observed using heat-inactivated HSV-1. The results suggest that resistance to HSV-1 is induced rapidly following immunization with viral antigen but that protection against encephalitis is independent of the cytokines that are generated in the peritoneum.

Innate and acquired immunity to herpes simplex virus type 1.

Immunization with heat-inactivated herpes simplex virus type 1 (HSV-1) 2-5 days before ocular infection reduced the frequency of establishment of latent HSV-1 infection in the trigeminal ganglion (TG); this induction of resistance coincided with reduced expression of IFN-gamma mRNA in the TG. Immunization with unrelated antigens was not protective. In part, this resistance to nervous system invasion correlated with the appearance of serum antibody to HSV-1. Immunization reduced viral replication in the eye and trigeminal ganglion, and prevented HSV-1 spread to the cerebellum. IFN-gamma was detected in immunized mice 4 days postocular infection as determined by plaque reduction using neutralizing Ab to IFN-alpha/beta and IFN-gamma. Injection of antibody (Ab) to IFN-alpha/beta and IFN-gamma administered at the time of immunization did not affect survival. Anti-IFN-gamma-treated mice had significantly reduced levels of IFN in their serum. Treatment with anti-IFN-alpha/beta Ab resulted in an elevation in viral replication as determined by the expression of latency associated transcripts in the TG of mice. Likewise, there was a significant increase in the CD8, IL-12 (p40), and TNF-alpha mRNA levels in the TG of the anti-IFN-alpha/beta-treated mice TG explant cultures demonstrated that viral load was significantly increased in the TG of anti-IFN-alpha/beta-treated mice relative to TG of control mice 7 days after infection. The results suggest that exposure to viral antigens 2-5 days before infection is an important determinant of the extent of HSV-1 spread to the nervous system. Moreover, the data suggest that both an antibody response and IFN-alpha/beta play a role in limiting the progress of infection from the peripheral tissues to the central nervous system.

[Biochemical and ultrasonic screening of chromosomal aneuploidies in the second trimester of pregnancy]. / Kromoszomális aneuploidiák biokémiai és ultrahangszürése a terhesség második trimeszterében.

For prenatal screening of chromosomal aneuploidies (primarily the most frequent Down syndrome) maternal serum AFP screening in the second trimester of pregnancy has been supplemented by the determination of hCG in Hajdú-Bihar county. In pregnancies at risk on the basis of biochemical tests, a thorough, aimed ultrasound examination for the detection of minor and major anomalies characteristic for chromosomal abnormalities was carried out. If both biochemical and ultrasound examinations were suggestive of high risk prenatal karyotyping was offered. During a two-years prospective study 14328 pregnancies were screened. Authors could detect 38% of Down-syndrome cases prenatally, 5 cases in pregnant women at age under 35 years and at the same time the number of amniocenteses increased only by 70. It was concluded that 66% of all Down syndrome cases could have been prenatally diagnosed if prenatal chromosome test were performed in all cases at high risk on the basis of screening tests and maternal age.

Ventriculomegaly with radial and renal defects: prenatal diagnosis in two consecutive sibs.

We describe two consecutive mid-trimester fetuses of different sexes with identical anomalies of the upper limbs and the kidneys in association with severe ventriculomegaly. We compare this apparently autosomal recessive syndrome to VACTERL-H association, Fanconi anemia, and two other, so far unparalleled syndromes. Taking into account the absence of chromosome breaks, the associated changes of the amniotic fluid, and the renal histology, we conclude that we are dealing with a different entity.

Impact of prenatal mid-trimester screening on the prevalence of fetal structural anomalies: a prospective epidemiological study.

The objective of this study was to evaluate the effectiveness of the measurement of maternal serum alpha-fetoprotein (MSAFP) at 16 weeks and a subsequent routine ultrasound screening at 18-20 weeks' gestation and the impact on the birth prevalence of congenital structural anomalies in an unselected pregnant population of Hungary in a prospective epidemiological study. A total of 63,794 pregnant women (representing one-sixth of the population of Hungary) were offered this screening program over 3 years (1988-90). Of the pregnant population, 75.7% (48,312) received MSAFP screening and in 81.0% (51,675), at least one ultrasound scan was performed. In the screened pregnancies, 496 craniospinal, thoraco-abdominal, urogenital and other severe major anomalies occurred; 317 were detected at 18-20 weeks (sensitivity 63.1%; specificity 100.0%; positive predictive value 100.0%). The sensitivity of ultrasound scanning was significantly higher (p < 0.05) than that of the MSAFP screening. (At the time of ultrasound scanning the MSAFP value was known.) In this study, the less serious anomalies such as hydrocele, hypospadias and undescended testicle were not systematically searched for, but the birth prevalences were calculated. The overall mid-trimester prevalence of severe plus less severe major anomalies was 2.26%. The birth prevalences of severe major anomalies were 0.57 (craniospinal), 4.36 (thoracoabdominal and urogenital) and 1.21 (other severe) per 1000. These values were lower than the mid-trimester prevalences which were 2.94, 5.20 and 2.06 per 1000, respectively. The prevalence values at the age of 1 year were also calculated (0.36, 2.21, 0.54 per 1000, respectively). We conclude that our screening program with availability of termination of pregnancy could significantly (p < 0.05) reduce the prevalence of severe major abnormalities at birth. Training programs in cardiac scanning are required.

Maternal age-dependent and sex-related changes of gestational serum alpha-fetoprotein.

Maternal serum alpha-fetoprotein (MSAFP) concentration values were measured in relation to maternal age and fetal sex in the 16th to 20th gestational weeks in samples taken from 9,556 pregnancies with the outcome of live, mature and healthy infants. Our results show positive significance between MSAFP concentration and maternal age (p < 0.001); we also found significantly higher AFP values in male fetuses than in female fetuses (p < 0.001). This specificity is by all probability due to the change in the physiologic AFP concentration of the pregnant woman. Considering maternal age and the new percentile AFP values, cut-off concentration values can be corrected, thus the quality of routine AFP screening can be improved. At the same time, with the application of the above parameters, a more effective selection of pregnancies at high risk for Down syndrome can be achieved.

[Screening for congenital anomalies in mid-term pregnancy. Prospective epidemiologic study. Fetal trisomy]. / Congenitális anomáliák szürése a terhesség középsö harmadában. Prospektív epidemiológiai tanulmány. Magzati trisomiák.

The authors have presented their experiences on prenatal screening of fetal trisomies in this second part of a prospective study between 1988 and 1990. They gained their conclusions by processing 63,496 pregnancies during three years. The results show that maternal age plays the most important role in the prenatal screening of fetal trisomies in Hungary. They recommend fetal karyotyping for every pregnant woman aged 35 years or more. They emphasized that using of a combined screening method (i.e. maternal age, serum alpha-fetoprotein, human chorionic gonadotropin, oestriol) is only permissible if the hormonal and cytogenetic laboratory background are provided under standard circumstances. Since these are not available for the vast majority of pregnant women in Hungary they concluded that, at least for the time being, the main criteria for prenatal screening of fetal trisomies is the maternal age. By applying this recommendation 25-30 percentage of Down syndrome fetuses can be detected.

[Possibilities for modifying risk factors for chromosome abnormalities--advantages of the so-called "triple" marker studies in comparison with pure "maternal age screening"]. / Möglichkeiten der Modifizierung der Risikoangaben für Chromosomenstörungen--Vorteile der sog. "Triple"-Marker-Untersuchungen im Vergleich zum reinen "Altersscreening".

We have offered the so-called "triple-marker screening" since May 1991 to all patients who came for prenatal care and did not select an invasive procedure primarily. First evaluation of 5210 cases revealed that 3.7% were test-positive for neural tube defects, 13.8% for Down's syndrome and 0.5% for both at the same time. The explanation for the comparatively high "test-positive" rate of 13.5% is the maternal age distribution with the median at 31.4 years. The highest number of women selecting triple-marker determinations was in the age group of 35 years. We detected 16 cases of Down's syndrome and in this group the majority of women was below 35 years. The decision of women to have an invasive procedure was obviously very much influenced by the actual risk assessment, because amniocentesis was chosen by 72/85 (84.8%) of women with a risk of more than 1:50, 192/290 (66.2%) of women in the risk category of 1:51 to 1:200 and 182/333 (54.6%) in the risk category of 1:201 to 1:400. The follow-up is not yet complete, but there is already good evidence for the efficiency of the screening program. Triple-marker screening also proved predictive in 10 cases of trisomy 18 and 8 cases of triploidy in this series. As a cut-off value we chose the risk of 1:386 which is equivalent to the odds of a 35 year old to have a child with Down's syndrome at birth.(ABSTRACT TRUNCATED AT 250 WORDS)

[Screening of congenital anomalies in the mid-trimester of pregnancy. Prospective epidemiologic study. Developmental anomalies]. / Congenitalis anomáliák szürése a terhesség középsö harmadában. Prospektív epidemiológiai tanulmány. Fejlödési rendellenességek.

Authors report about the data of 60,000 pregnant women from three district counties of Eastern-Hungary between 1988 and 1990, on whom both ultrasound and alfa-fetoprotein screenings were performed. They demonstrate the value of this screening for the detection of fetal malformations and for the birth prevalence rate as well. This effective screening-protocol is proposed for a nation-wide application.

[Retrospective survey of nursery-age children 5 years after Chernobyl]. / Ovodások retrospektív felmérése Csernobil után 5 évvel.

In their paper, the authors are dealing with the effect of the Chernobyl's nuclear catastrophe (1986) for the outcome of pregnancy and some parameters of the status of health. Their empirical study is based on a sample (sample size is 1168) of pregnant of the critical year 1986, and also based on a control sample (sample size is 1068) of the year 1985. They compared the five old year children's parameters of status of health between the critical and control groups. Their retrospective epidemiologic survey is extended to study some prenatal, perinatal and childhood parameters also. Finally, they summarise the biometric evaluation of their questionnaire survey. No significant difference of health conditions of the two groups defined above was found.

[Oligohydramnios in mid-term pregnancy: analysis of 182 cases]. / Oligohydramnion középidös terhességben: 182 eset elemzése.

The outcome and pathological background of 182 pregnancies with mid-trimester oligohydramnios are discussed. Maternal serum alpha-fetoprotein (AFP) concentration in the 16th week of gestation was also determined in 119 cases. MSAFP in pregnancies with oligohydramnios associated urinary tract malformations was found to be mostly in the normal range, but it is often elevated in the cases without malformation. In addition, normal AFP was found in most cases, where the newborns survived the perinatal period. It can be concluded, that the elevated maternal serum AFP without ultrasonically detectable malformation refers to the extrafetal origin of the oligohydramnios, and it is recommended to take it into consideration in the genetic counselling practice.

Prenatal diagnosis of cystic fibrosis by microvillar membrane enzyme analysis in amniotic fluid.

Prenatal diagnosis was performed in 92 pregnancies high-risk for cystic fibrosis during six years. Amniotic fluid samples obtained by amniocentesis were examined with regard to their microvillar membrane enzyme activity. Though trehalase, alkaline phosphatase isoenzymes and L-gamma-glutamyltransferase in the amniotic fluid are not specific markers of cystic fibrosis, their activity is significantly lower than in normal pregnancies. By measuring the three enzymes simultaneously, sensitivity, specificity and reliability of the method were found to be over 92%. It is concluded that mid-trimester amniotic fluid diagnosis is indispensable for some heterozygotic couples for cystic fibrosis even in the possession of DNA (desoxyrobonucleic acid) methods.

Discriminant analysis for assessing the value of amniotic fluid microvillar enzymes in the prenatal diagnosis of cystic fibrosis.

We have analysed the sensitivity, specificity, and reliability of biochemical diagnosis based on microvillar membrane enzyme assay and using discriminant analysis in amniotic fluid samples obtained from 54 pregnancies at high risk for cystic fibrosis and 125 normal pregnancies. Our results show that amniotic fluid trehalase, alkaline phosphatase, alkaline phosphatase isoenzymes and gamma-glutamyltransferase enzyme activities measured during 16-20 gestational weeks, in spite of their non-specificity for cystic fibrosis, have a very good predictive value for fetal cystic fibrosis or exclude the possibility of the disease. Overall enzyme activity analysis provided over 90 per cent reliability of the method.

Amniotic fluid microvillar enzyme activity in fetal malformations.

Prenatal diagnosis of cystic fibrosis based on amniotic fluid microvillar enzyme activity assay has become routine practice in the past few years. Normal (median) values of these enzymes were determined in 177 normal healthy pregnancies between 15-20 gestational weeks and were related to enzyme values measured in 50 pregnancies complicated with congenital malformations, 6 monogenic inherited diseases and 4 chromosomal aberrations. It is concluded that increased trehalase activity has diagnostic importance in detecting fetal kidney diseases, and radial-renal syndrome (with elevated GGT activity), while low enzyme activities may indicate chromosomal aberrations (with no signs of intestinal obstruction). With the collection of further data, the analysis of these enzymes might provide an opportunity to set up diagnostic procedures for the detection of other, non-CF-related cases.