Vibrational heat capacity of silver carp collagen.

The study of the experimental and calculated heat capacity, Cp of fish collagen (silver carp) with contents of several additive components was presented. The experimental low-temperature heat capacity was measured in the temperature range of 1.85 to 302.8 K using a Quantum Design Physical Property Measurement System (PPMS) and the higher temperature Cp from 223.15 K to 382.15 K by Differential Scanning Calorimetry (DSC) method. For an interpretation of the experimental, low-temperature data, the vibrational heat capacity of the pure silver carp collagen was calculated based on the contribution of a sum of the vibrational heat capacity of 4248 amino acids. The vibrational heat capacity for each amino acids was taken from Advanced Thermal Analysis System (ATHAS) Data Bank for individual poly (amino acid) residues based on their group and skeletal vibrational spectra. Comparing of the experimental heat capacity of the collagen with additive components and the calculated vibrational heat capacity of the pure silver carp collagen shows that the differences range from around 10% at 100 K to 14% at 300 K temperature. Such thermal analysis can provide information about the contribution to Cp of unknown components or impurities in the investigated system.

N-Acetyl-ß-D-hexosaminidase in gestational diabetes mellitus - a preliminary study.

PURPOSE: N-Acetyl-ß-D-hexosaminidase (HEX) is an exoglycosidase which has been extensively studied and which has been used as a marker for inflammation. It was therefore thought that measurement of the activity of this enzyme might be useful in diagnosing gestational diabetes mellitus (GDM) as this condition is frequently associated with inflammation. The main object of the study was the determination of N-acetyl-ß-D-hexosaminidase activity in women with GDM and 3 months postpartum in comparison with control groups of non-pregnant and healthy pregnant women. MATERIAL AND METHODS: Twenty-five blood serum samples from women with GDM and women 3 months postpartum; 20 blood serum samples from non-pregnant and healthy pregnant women (control groups) were enrolled into the study. Serum was prepared from all blood samples and HEX activity was measured by the method of Chateriee et al. (modified by Zwierz et al). RESULTS: A statistically significantly increase in the activity of HEX in the GDM blood serum was found as compared to the control groups (p<0.05). Further analysis showed a statistically significant decrease in the activity of HEX among postpartum women, but the level of enzyme activity was still above the normal control level in comparison to the control group of nonpregnant healthy women (p<0.05). CONCLUSIONS: Changes in the activity of HEX appear to be involved in the pathogenesis of gestational diabetes mellitus. Determination of HEX activity may have prognostic significance as an early indicator of diabetes mellitus among GDM women in the future.

Evaluation of interleukin concentrations in amniotic fluid in preterm and term parturition and in oligohydramnios.

BACKGROUND: The goal of our work was to evaluate the activity of interleukin 1 beta, 6, 8, 12, and 18 in the amniotic fluid in preterm and term deliveries, and to specify the dependency between the amniotic fluid index (AFI) and the concentration of these interleukins. MATERIAL AND METHODS: Amniotic fluid (27 samples from preterm deliveries and 49 from term deliveries) was collected during cesarean section or in the course of amnioinfusion performed when the pregnancy was accompanied by oligohydramnios (AFI< or = 5cm). The amniotic fluid thus obtained was centrifuged at 1000 xg for 15 min. at a temperature of 4 degrees C, and the filtrate was kept at a temperature of -80 degrees C until assayed. The assays were done by the ELISA method, using ready antibody kits from 'R&D Systems'. A comparative analysis was done for the concentrations (expressed in pg/ml) of the individual interleukins found in the amniotic fluid from preterm and term deliveries, and for the amniotic fluid index (AFI< or = L5 and >5). The statistical analysis was performed using the Wilcoxon test. RESULTS: The increased level of IL-12 we found in the amniotic fluid from preterm deliveries and those complicated by oligohydramnios may indicate the participation of immunological mechanisms in the pathogenesis of oligohydramnios.

[Prenatal and neonatal diagnosis of osteogenesis imperfecta in obstetrical practice]. / Diagnostyka przed i pourodzeniowa wrodzonej lamliwosci kosci (Osteogenesis imperfecta) w praktyce polozniczej.

Osteogenesis imperfecta (OI types I, II, III, IV) is a heterogeneous group of genetically disorders of connective tissue. Quantitative or qualitative abnormalities of type I collagen form pathogenetical basis of the disease. They are caused by mutations in genes encoding collagen proteins or enzymes involved in collagen biosynthesis. The clinical features of each type usually correspond to the type of mutation. Typical manifestations are fragile bones with multiple bone fractures and bone deformities. Currently applied diagnosis in utero of OI II and sometimes OI III may be performed. Diagnosis of other OI phenotypes cannot be made until after birth. We present three cases of OI II (four children) diagnosed, in utero, by ultrasound examination. The analysis in work include: 1. the prenatal sonographic features of OI type II 2. the biochemical properties of collagen in the above cases 3. genetic counselling of the families affected by OI.

[Relationship between Doppler velocimetry at middle cerebral artery and umbilical artery and status of newborn]. / Relacja miedzy parametrami dopplerowskimi przeplywu krwi w tetnicach srodkowej mózgu i pepowinowej a stanem noworodka.

Aim of investigations was qualification of account between Doppler parameters in estimation of fetal state. Investigations one passed on 30 fetuses and newborn children in pregnancies brought. Doppler parameters one priced at use of sonographic device Toshiba SSH 140 A/G and searchers of type convex about working frequency 3.75 MHz. Following Doppler flow blood parameters were analyzed: maximum blood speed (V1) average blood speed (V2) and minimum blood speed (V3), systolic/diastolic ratio (S/D), resistance index (RI), pulsatile index (PI) and proper flow in umbilical cord vein (MF) in following dishes of feto-placental circulation: middle cerebral artery (MCA) and umbilical cord artery (UA). Acid-base equilibrium and gasometry of blood in umbilical cord dishes one marked at use of device Ciba-Corning 278 Blood Gas System and parameters of oxygenation of blood at use of device Ciba-Corning 270 CO-OXIMETER. At new-born children one priced pH-metry (pH) and gasometry (pO2, pCO2, BAA) in blood umbilical cord arterial and venous were measured. The newborn children were estimated by Apgar score. There were following essential statistical correlations between Doppler parameters of fetal blood flow and with parameters of acid-base equilibrium of new-born child: 1/ between V2 and V3 in UA and with supply of rules (BAA) in UV (p = 0.027; p = 0.009) and UA (p = 0.035; p = 0.003) and venous pH (p = 0.022; p = 0.009); 2/ between RI in UA and BAA in UV (p = 0.006) and UA (p = 0.010); 3/ between PI in UA and BAA in UV (p < 0.0001) and UA (p < 0.0001) and pH venous (p < 0.0001). We can conclude that Doppler investigations only by measure of parameters of blood flow in middle cerebral artery and umbilical artery in expectation of state of birth new-born child priced across parameters of acid-base equilibrium and Apgar score are not very useful, however they are helpful.

Doppler examinations in the prognosis of birth status of the newborn.

AIM OF THE STUDY: Determination of correlation between Doppler parameters in the evaluation of fetal state and biochemical and clinical determinants of birth status of the newborn. MATERIAL: 30 full-term pregnancies. METHODS: Doppler parameters were determined in maternal vessels (uterine and arcuate arteries), fetal vessels (middle cerebral arteries, renal artery and descending aorta) and in umbilical arteries and veins. Levels of uric acid, xanthine and hypoxanthine, which reflect oxygen deficiency, were assessed in amniotic fluid by means of HPLC method. Furthermore, clinical condition of a newborn (rating according to the Apgar scoring system), pH-metry and gasometry in umbilical blood were assessed. RESULTS: Significant correlation between determinants of fetal and neonatal condition and Doppler parameters were determined, that is: between uric acid concentration and blood flow in fetal cerebral arteries, between base excess, pH and blood flow in umbilical vessels, between clinical examination of birth status of the newborn and renal and arcuate arterial flow and between pO2 and S/D ratio in renal artery. CONCLUSIONS: On the basis of this correlation, the following conclusions were drawn: Doppler examinations are highly useful in the prediction of birth status of the newborn if the specific conditions and appropriate range of the examination are maintained, i.e.: 1) examination must be multivascular, 2) it should involve cerebral artery, parenchymal resistance vessels (for example renal artery), umbilical artery and vein, intramural uterine vessels (for example arcuate or spiral artery), 3) among blood flow parameters not only standard indices (S/D, RI, PI), but also arterial blood flow waves velocity, specific flow and transverse section field should be considered.

Uterine and fetal Doppler flow changes after misoprostol and oxytocin therapy for induction of labor in post-term pregnancies.

OBJECTIVES: To compare the effect of misoprostol vs. oxytocin on blood flow in uteroplacental circulation during labor induction. METHODS: Ninety-one women with indications for induction of labor were assigned to receive either misoprostol 50 microg per vagina every 4 h as needed or intravenous oxytocin by means of a randomization table generated by computer. Doppler velocimetry of umbilical, uterine and arcuate arteries was performed immediately before and 2-3 h after administration of misoprostol or oxytocin. Pulsatility index (PI), resistance index (RI) and systolic/diastolic (S/D) ratios were measured for these arteries. The SAS system was used to perform statistical analysis. RESULTS: There were no significant changes of PI, RI and S/D ratios in umbilical arteries during misoprostol and oxytocin inductions. Vaginal application of misoprostol significantly increased PI, RI and S/D ratios in arcuate arteries and S/D ratio in uterine arteries. CONCLUSIONS: Our results indicate that intravaginal misoprostol administration increases uteroplacental resistance but probably does not substantially affect placental perfusion.

[Antepartum amniotic fluid index and pregnancy outcome]. / Przedporodowy indeks plynu owodniowego w przewidywaniu wyniku ciazy.

Our purpose was to evaluate prognostic value of antepartum amniotic fluid index (AFI) on fetal and newborn outcome, because oligohydramnion is associated with increased risk of fetal and newborn distress. Perinatal outcome was defined by 1' and 5' Apgar score, intrapartum fetal heart rate (FHR), blood gas, acid-base status in umbilical artery, mode of delivery and meconium-stained amniotic fluid. Data was analysed with CSS Statistica for Windows package by test t for independent samples and chi 2. Prospective observations was conducted with 84 singleton pregnancies complicated by oligohydramnion (AFI < or = 5 cm). A 336 pregnant women, with normal amniotic fluid index and no complicated pregnancy, was considered as controls. No increased operative delivery and meconium-stained amniotic fluid rate was noted. There was no significant effects AFI on blood gases in umbilical artery and Apgar score. Oligohydramnion was associated with increased risk of fetal acidosis (pH < 7.16) (p = 0.009) and abnormalities of FHR (variable decelerations) (p = 0.004). Reduced AFI had poor predictive value of fetal acidosis and newborn depression positive predictive value (8-13%). Summarizing the results, our investigation showed that amniotic fluid index is only one point in the prognosis of perinatal outcome.

Turnip yellow mosaic virus isolates with experimentally produced recombinant virion proteins.

The consensus secondary structure of the virion proteins (VPs) of tymoviruses was predicted from their amino acid sequences using a combination of computer methods: profile alignment, hydrophobicity moment and 'PredictProtein'. All methods predicted that they were eight-stranded anti-parallel beta-barrels with two alpha-helical regions. The predicted structure was used to design recombinants of turnip yellow mosaic virus (TYMV) in which selected parts of its VP were replaced with homologous regions of belladonna mottle virus (BeMV) in a cDNA clone encoding the genome of TYMV. Six of ten such recombinants were fully viable and most gave symptoms in Chinese cabbage indistinguishable from those of TYMV, although they did not always infect plants systemically and none infected hosts of BeMV or of other tymoviruses. A TYMV recombinant with the N-terminal part of its VP replaced with the E71 epitope of Plasmodium falciparum was also viable, but others with the same region replaced with the V3 region of the lentivirus human immunodeficiency virus type 1 were not. Epitope analysis of antisera prepared against the virions of parental TYMV and some of the recombinants showed that, although the N terminus of the VP is immunogenically dominant, it is not exposed at the surface of the virion, a finding confirmed by comparing the electrophoretic mobilities of the virions. The recently published structure of the TYMV VP determined by X-ray crystallography confirms the accuracy of the predicted secondary structure of the VP, and hence the utility of the methods used.

Genetic variation in populations of kennedya yellow mosaic tymovirus.

Kennedya yellow mosaic tymovirus (KYMV) occurs along the eastern Australian seaboard in the perennial legumes Desmodium triflorum and D. scorpiurus in the north, and Kennedya rubicunda in the south. The genetic variation of more than 100 isolates of KYMV, most of them from the north, has been studied using an RNA hybrid mismatch polymorphism (RHMP) method. The method clearly separated the isolates into two groups; all the northern Desmodium isolates formed one group and all the Kennedya isolates from the south another. These sub-populations were themselves variable and the Desmodium population alone was more variable than that of the related turnip yellow mosaic tymovirus in the Kosciusko alpine area.

The genomic sequence of cardamine chlorotic fleck carmovirus.

The complete genomic sequence of cardamine chlorotic fleck carmovirus (CCFV) has been determined. The genome is a positive-sense ssRNA molecule 4041 nucleotides in length, and has 47 to 64% sequence identity with turnip crinkle, carnation mottle and melon necrotic spot carmoviruses. CCFV and these other carmoviruses have four similar open reading frames (ORFs), and CCFV has large regions of amino acid identity in all of these ORFs with a European isolate of turnip crinkle virus. CCFV, which replicates well in Arabidopsis thaliana, has only been found so far in Australia in the wild perennial brassica Cardamine lilacina.

RNA hybrid mismatch polymorphisms in Australian populations of turnip yellow mosaic tymovirus.

In the Mt. Kosciusko alpine area of Australia there are three well-separated populations of Cardamine lilacina, an endemic sward-forming perennial brassica, and these are infected with turnip yellow mosaic tymovirus. The genetic variation in these viral populations has been assessed by an RNA hybrid mismatch polymorphism method. About 100 isolates were examined; the genomic RNA of each isolate was prepared from a shoot of a single wild C. lilacina plant. RNA hybrid mismatch polymorphisms (RHMPs) were assessed in six regions of the genomes using labelled negative-strand probes transcribed from selected portions of a cloned TYMV genome. The probed region at the 3' end of the genome showed little variation and over 95% of the isolates gave the same pattern. However, other parts of the genome, including the 5' non-coding region, were much more variable. There was no significant correlation between groupings based on the RHMP patterns, and the location from which the isolates were collected, nor with the symptom type or severity shown by their host plants. The patterns of variation suggested that all three populations of the virus are a single quasi-species; at most one tenth of the isolates gave similar RHMP patterns, those of the "master copy".

Infectious eggplant mosaic tymovirus and ononis yellow mosaic tymovirus from cloned cDNA.

Eggplant mosaic virus (EMV) and ononis yellow mosaic virus (OYMV) are two tymoviruses that have ssRNA genomes of about 6.2 kb and 6.3 kb, and which infect solanaceous and leguminous hosts, respectively. Full-length cDNA clones of these viruses were constructed with a T7 promoter adjacent to the 5' terminus of the DNA copy of the viral genome, and with unique restriction endonuclease sites at the 3' terminus. This allowed RNA to be transcribed from the DNA encoding the genome. The transcript RNA was infectious when inoculated to Nicotiana glutinosa (for EMV) and Pisum sativum (for OYMV). These clones, together with clones of turnip yellow mosaic tymovirus, which infects brassicas, have been used to construct hybrids in which the virion protein gene was exchanged between EMV or OYMV and turnip yellow mosaic virus. These and other hybrids are being used to investigate the molecular basis for host range differences in tymoviruses.

Turnip yellow mosaic virus variants produced from DNA clones encoding their genomes.

Full-length dsDNA clones that encode the genomes of two Australian turnip yellow mosaic isolates, TYMV-BL and TYMV-CL have been constructed. These clones were transcribed to give 6.3 kb capped ssRNA which infects Chinese cabbages to give symptoms indistinguishable from those produced by the parental viruses. Extensions of up to 26 nucleotides at the 3' end of the TYMV-BL clone delay infections, but virus particles isolated from these plants 4 weeks after inoculation contain RNA with the original TYMV-BL 3' terminus. A 90 nucleotide-long portion of the virion protein gene of TYMV-BL was replaced by a synthetic 90-mer primer with 16 nucleotide changes to decrease the large cytosine content (34-42%) characteristic of tymovirus genomic RNA. No reversion of any of the mutated nucleotides to cytosine occurred during 7 passages in Chinese cabbage. Hybrids between the TYMV-BL and TYMV-CL clones were also constructed, by exchanging various portions of the genome. However, it was not possible to determine definitively which part of the viral genome is responsible for the more severe symptoms caused by TYMV-BL as the hybrids gave intermediate symptoms.

Coronary artery disease and apolipoprotein A-I/C-III gene polymorphism: a study of Saudi Arabians.

The infrequent band of 3.2-kb of the apolipoprotein A-I/C-III polymorphic region has previously been found to be associated with coronary artery disease and with hypertriglyceridaemia in Caucasians. We studied the apolipoprotein A-I/C-III gene cluster polymorphism in 97 Saudi Arabians in relation to coronary artery disease. Patients were categorized as being with or without coronary artery disease on the basis of coronary angiography. Genomic blotting of Sac I-digested chromosomal DNA with the use of an apolipoprotein A-I gene probe revealed 4.2-kb and 3.2-kb hybridization bands. The genotype frequency of patients with and without coronary artery disease was not different. The frequency of the 3.2-kb allele occurred in 16% of patients with coronary artery disease and in 21% of patients with normal coronary arteries (non-significant). In conclusion, we have not been able to confirm in Saudi Arabians associations previously reported in Caucasians of the 3.2-kb band and coronary artery disease.

The tymobox, a sequence shared by most tymoviruses: its use in molecular studies of tymoviruses.

The 5'-terminal sequences of the virion protein mRNAs of ononis yellow mosaic and kennedya yellow mosaic tymoviruses were determined, and also the positions in the genomes of the transcription initiation sites of those mRNAs. Comparisons of the available genomic sequences of tymoviruses revealed two conserved regions, one at the initiation site and another longer sequence of sixteen nucleotides to the 5' side of it. The longer sequence, which we call the tymobox, was tested as a target for a designed ribozyme, which cleaved appropriate genomic fragments of three tymoviruses. A synthetic oligonucleotide with sequence complementary to the tymobox was shown to be a tymovirus-specific probe for diagnosing and identifying tymoviruses, except for wild cucumber mosaic tymovirus. The tymobox sequence was also used as a primer for the second strand DNA synthesis of dsDNA representing the virion protein gene of cacao yellow mosaic tymovirus, a tymovirus with unknown sequence. Thus, the tymobox is a useful tool in molecular studies of tymoviruses.

Apolipoprotein A-I/C-III gene cluster polymorphism in Saudi Arabians, Filipinos and Caucasians.

We studied the polymorphic locus in the A-I/C-III gene cluster on the long arm of chromosome 11 in 147 Saudi Arabian, in 84 Filipino and in 69 Caucasian blood donors. Digestion of DNA yielded two fragments 4.2 kb and 3.2 kb long. The genotype distribution was the same in Arabs and Filipinos, but both were significantly different from Caucasians (p = 0.005 and 0.0005). The 3.2-kb allele occurred in 18% of the Saudi Arabians, in 23% of the Filipinos and in 4% of Caucasians. The frequency of the 3.2-kb allele was significantly higher in Arabs and Filipinos compared to Caucasians (p = 0.0005).

Coronary artery disease, HDL cholesterol, and insulin-gene flanking sequences.

The relationship between coronary artery disease, HDL cholesterol, and the hypervariable region flanking the human insulin gene was studied in Saudi Arabian non-diabetic subjects (n = 68) and in patients with Type 2 diabetes (n = 35). A locus of insulin-gene-linked DNA polymorphism with three average size classes of alleles was found: a small class 1, a rare medium size class 2, and a large class 3 allele. In the total group of subjects (n = 103), those with the class 3 allele had a lower plasma HDL cholesterol concentration than those without the class 3 allele (0.93 +/- 0.26 vs 1.12 +/- 0.30 mmol l-1, 2p less than 0.003). No difference in genotype and allelic frequency was found between patients with and without coronary artery disease in the combined group of subjects, with and without diabetes. Similarly the genotype distribution was not different between non-diabetic subjects and patients with Type 2 diabetes in the combined group of subjects, both with and without coronary artery disease. In conclusion, the study did not confirm a previous study showing an association between the class 3 allele and atherosclerosis in a Caucasian population. However, the class 3 allele was associated with a low plasma HDL cholesterol concentration.

Incidence and transfer of R-plasmids at a hospital in Saudi Arabia.

One hundred and fifty Gram-negative bacteria isolated from patient specimens at King Faisal Specialist Hospital were examined for their ability to transfer antibiotic resistance plasmids to a sensitive Escherichia coli recipient in conjugation and transformation experiments. Agarose gel electrophoresis was used to enumerate and size the R-plasmids found, and Southern DNA hybridization was used to assess similarities between antibiotic resistance plasmids from different bacteria and sources. Of the bacterial isolates tested 65% contained plasmids, 70% of these transferred antibiotic resistance to E. coli, and 40% transferred multiple, linked resistances on R-plasmids. DNA hybridization of these R-plasmids demonstrated widespread similarities between plasmids from different bacterial genera and from different hospital locations. In particular, a gene encoding ampicillin resistance appeared especially widespread, indicating that a transposon may be mediating transmission of this resistance.