Multiplex RT-PCR-ELISA compared with bioassay for the detection of four apple viruses.

A sensitive and reliable multiplex RT-PCR-ELISA technique for the detection of Apple chlorotic leaf spot virus, Apple stem pitting virus, Apple mosaic virus and Apple stem grooving virus was developed. This technique is compared with the method used commonly for indexing by woody indicators, which is time consuming and expensive. For the RT-PCR-ELISA technique, the amplified products were labeled with digoxigenin during the RT-PCR by incorporation of a digoxigenin labeled primer. After hybridization of the PCR products to specific capture oligonucleotides, which were bound covalently to the surface of NucleoLink strips, anti-digoxigenin antibodies were used for detection. More than 100 samples were tested in parallel by indexing and multiplex-RT-PCR-ELISA. All infections detected by woody indicators were also detected by multiplex RT-PCR-ELISA. Furthermore, additional infections were only found by multiplex RT-PCR-ELISA. The colourimetric detection of multiplex-RT-PCR products was at least as sensitive and sometimes slightly more sensitive than detection by gel electrophoresis. The results show that this molecular technique is more reliable for the detection of the above mentioned apple viruses than indexing by woody indicators, thereby helping to reduce cost and time during the certification of plant material.

[Circadian rhythm of pregnancy contractions]. / Circadiane Rhythmik von Schwangerschaftskontraktionen.

Continuous measurement of normal contractions in 57 primiparae and multiparae showed a minimum in the mornings and a marked peaking of frequency between 8:30 PM and 2:00 AM. This could be due to a periodic fluctuation in the parameters influencing uterine motility. At night, motility-enhancing factors such as estrogens and prostaglandins predominate. These results suggest that in cases at high risk for premature delivery, with high pelvic scores, therapy should be continued at night. In addition, the time when the contractions were recorded should be taken into account when assessing their frequency, and the measurement repeated if necessary.

[Waste prevention and recovery of valuable raw materials in the hospital]. / Abfallvermeiden und Wertstoffsammeln im Krankenhaus.

Quite a number of the diseases with which Public Health services are concerned are due to environmental factors. It is our aim that especially the Public Health staff should tackle with efficiency and dispatch all the problems connected with damage caused by such factors. Environmental protection is in fact Medical Science. The volume of the waste deposits has been exhausted to the full. Burning of refuse is not the proper alternative: natural science tells us that it is definitely no solution of the problem. The best kind of waste is not to waste anything, i.e. waste that does not arise at all. The belief that disposable or throwaway goods improve hygiene, is totally erroneous and a misconception. All persons engaged in Public Health must unanimously demand from industry the production of high-quality, well sterilisable and easy-to-clean package items for repeated use. Active environmental protection motivates the staff and enhances the reputation of their work.

[The Cerviport--a device for the continuous measurement of the dilatation of the ostium uteri during labor]. / Cerviport--ein Gerät zur kontinuierlichen Messung der Muttermundsweite unter der Geburt.

In the past few years obstetrics has experienced substantial improvements due to the apparatus supervision of the fetus. What remained an unsolved problem was the continuous measuring of the dilatation of the cervix in labor, although various methods have been developed. The forceps given by Richardson et al. 1978 have been further developed by us. These cervical forceps are attached to the cervix at 3 and 9 o'clock. The patulousness of the cervix corresponds to the angular aperture of the forceps and is directly converted into an electric signal by means of a dilatation-measuring strip attached to the branches of the forceps and said signal is continuously recorded by a compensation writer which is synchronized with the cardiotocograph. 110 women between the 34th and the 41st week of pregnancy were examined. In the course of the measurements carried out, the application of the cervical forceps became easier for the obstetrician, because he acquired more and more routine in applying such forceps. The indication for cervical measuring includes delayed labor, premature rupture of the fetal membranes, the fetus letalis ante partum as well as the attempt of a spontaneous partus after a Caesarean section in combination with measuring the intrauterine pressure. Because no increased danger of infection was observed, cervical measuring is considered to be an enrichment in the field of obstetrics. The inconveniences caused to the patient by applying the cervical forceps are by far made good for by the absence of manual examinations. Hence, one can assume that cervical measuring in the case of risk patients during pregnancy constitutes a useful additional measure.

[Holoprosencephaly--clinical picture and genetic counseling in 6 cases]. / Holoprosenzephalie--Klinisches Erscheinungsbild und genetische Beratung in sechs Fällen.

Inadequate midfacial and forebrain development may result from constitutional chromosome aberrations, gene defects and, possibly, from as yet unidentified terato-genetic agents. The spectrum of clinical manifestations ranges from cyclopia with grossly incomplete organogenesis of the forebrain to apparently minor deviation from normal midface morphogenesis presenting as hypotelorism or absence of the philtrum. Such minor facial dysmorphias may, however, be likewise accompanied by severe anomaly in brain development and function. We report six cases of holoprosencephaly defects in children without demonstrable chromosomal anomalies. The presenting clinical symptoms in these cases were anomalies of cranio-facial shape, hypotelorism; nasal and ocular malformations, as well as median clefts. Some cases presented additional defects in extra craniofacial regions. Two infants who survived for several hours showed evidence of forebrain defects on CT-scans. Three of the cases suggest autosomal-recessive inheritance with 25% recurrence risk on the basis of proven or highly probable parental consanguinity. The remaining, presumably sporadic cases carry a low empirical recurrence risk. Three of the six cases received direct or indirect hormone treatments during early pregnancy.

Uterine contractions during pregnancy.

The principal function of muscular tissue is contraction. This is also true for the uterus this function begins with sexual maturation. During pregnancy special conditions are brought about by the increasing distention of the uterine cavity. For clinical practice, external tocography is of importance for the recognition of uterine motility Data in the literature on the normal frequency of contractions during pregnancy vary greatly and caused us to study this topic. Moreover, most cases of prematurity independent of other causes suppose a prematurely increased uterine motility. In order to register this activity a portable recording apparatus was developed. We studied in particular the question for how long and how often uterine contractions should be recorded. A one hour daily recording was found to be sufficient. In order to gain information on the average number of contractions it suffices to record during one week. In 26 healthy primiparous and 28 multiparous women the number of contractions from the 25th to 41st gestation week was recorded. A notable finding was the frequency peak about the 32nd week. The number of pregnancy contractions as a sole parameter for the recognition of impending premature delivery is not sufficient. However, the information may be important when taking in conjunction with the previous history and the state of the cervix. We considered tocolysis indicated when as a weekly average there are more than two contractions per hour with beginning cervical dilation or more than three contractions per hour regardless of cervical findings.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effect of exercise during pregnancy on uterine contractions]. / Uber die Einwirkung der Schwangerengymnastik auf die Schwangerschaftskontraktionen.

External tocometry was performed one hour before and one hour after pregnancy gymnastics in 102 healthy pregnant women between 25th and 38th gestational week. There were no statistically significant differences in the number of contractions. The short-term increase in frequency of contraction was without any clinical importance. No prolongation of the duration of contractions could be observed with advanced pregnancy. 34 per cent of the women were delivered in the 41st gestational week, 32 per cent before the 40th one. In case of threatening premature delivery gymnastics are contraindicated with exclusion of relaxation exercises. They are allowed only in case that a good attitude of contractions before and after gymnastics can be demonstrated by objective external tocometry.

[Antibiotic prevention in gynecology. Concentrations of ceftizoxime in the serum, myometrium, endometrium and fallopian tubes]. / Antibiotikaprophylaxe in der Gynäkologie. Konzentrationen von Ceftizoxim in Serum, Myometrium, Endometrium und Tuben.

34 patients received an intravenous bolus injection of 2 g ceftizoxime over 5 min at various times before abdominal or vaginal hysterectomy. Ceftizoxime concentrations in myometrium were substantially higher than in salpinges and endometrium. Tissue concentrations of 4 micrograms/g could be maintained for at least 3 hours. The ceftizoxime tissue concentrations attained in myometrium, endometrium and salpinges justify therapeutic studies and controlled prospective clinical trials on the prevention of postoperative wound infections with this antibiotic.

Registration of contractions in pregnant women using a microprocessor controlled digital recorder.

As a further development of the two component contraction recorder TOKOPORT, a digital microprocessor controlled contraction memory has been built which registers, analyses and displays contractions in pregnant women. The memorized data can be read out in graphic form. The possibility of conveying the data by telephone is under development. The device is small, simple to operate and portable.

Clenbuterol--a long term uterine relaxant.

Clenbuterol (C.) is a bronchospasmolytic agent which selectively stimulates beta 2-receptors; side-effects of the drug are virtually minimal. C. effects a strong uterine relaxation. The renal elimination after oral administration is 34 hours. In a pilot study 26 women during labor received C. because of uterine hypermotility and increased fetal risk. After oral doses of 40 microgram, 60 microgram and 100 microgram a marked uterine relaxation was achieved in 20 women. Six women did not respond, because the dilatation of the uterine orifice was too much progressed. The effect of an intravenous injection was not superior to that of an oral administration. 21 women with signs of imminent premature birth received C. orally during pregnancy. There were a number of high risk cases among these women. In average a period of uterine relaxation of 63 days was achieved. 18 women reached the 38th week of pregnancy. No cardiovascular side effects were noted in the fetuses, only at the begin of the treatment the mothers showed mild transient palpitations or also transient tremor.

[Pregnancies after ineffective interruption or intrauterine procedure in early pregnancy (author's transl)]. / Schwangerschaftsverlauf nach misslungener Abruptio oder intrauteriner Manipulation in der Frühschwangerschaft.

We report about 13 pregnancies after intra-uterine procedure in early pregnancy. In 12 cases infant and placenta were without any lesions. One infant had a traumatic loss of the left arm and shoulder caused by the ineffective interruption of pregnancy. Into discussion come the conditions in which we can support the wish to carry on pregnancy after an early intra-uterine procedure.

[On the feto-maternal transfusion of red blood cells during transplacental and para-placental genetic amniocenteses (author's transl)]. / Uber die Einschwemmung fetaler Erythrozyten bei para- und transplazentarer Amniozentese in der Frühschwangerschaft.

The concentrations of fetal hemoglobin (HbF) containing red blood cells in the peripheral blood of the mothers was studied in 113 cases by the method of Kleihauer & Betke prior and following trans-abdominal amniocenteses for genetic reasons between 15 and 18 weeks of pregnancy. Prior to the amniocentesis, 25% of the patients had HbF positive cells. Following the amniocentesis 36% of the patients showed HbF positive cells. In one out of six amniocenteses a measurable increase of the concentration of HbF containing red blood cells by approximately 0.22% was detected. These increases are within the limits found prior to amniocentesis and are not significant. There was no difference in the incidence and level of microtransfusions between paraplacental and transplacental amniocenteses. Because of the possibility of Rh sensitization Rh negative mothers should receive Rh immunoglobulin. None of the 112 patients had an abortion within two weeks following the amniocentesis.

Prenatal diagnosis of beta-thalassemia.

Prenatal diagnosis was attempted in 14 fetuses at risk for homozygous beta-thalassemia, gestational age 18-22 weeks. In 4 cases the placenta was entirely anterior, placental aspiration under ultrasonic control only had to be used for fetal blood sampling. In 10 fetuses fetoscopy was used for puncture of a chorionic blood vessel. Diagnoses were based on the rate of in vitro synthesis of beta-globin related to total non-alphaglobin synthesis. With the aid of fetoscopy, nearly pure fetal blood was obtained in general. Placental aspiration resulted in samples which contained a low percentage of fetal and a high percentage of maternal cells. The attempt of fetal blood sampling resulted in fetal loss in two cases. In 2 aspiration cases no diagnosis could be made because the samples were inadequate. In 2 cases the diagnosis was established in spite of low fetal cell content through determination of the specific radioactivity in the placental and pure maternal blood. Until now 6 children have been born in whom prenatal diagnosis had been attempted, none of them has homozygous thalassemia. Present efforts are directed toward improving the safety of fetal blood sampling and the biochemical methods for the diagnosis in placental samples with low fetal cell content. Although the prenatal diagnosis of beta-thalassemia is possible, the procedure has still to be considered experimental.