Group B streptococcus and pregnancy.

Group B streptococcus is a significant pathogen for both mother and child. routine urine culture in pregnancy will identify and allow treatment of women with asymptomatic bacteriuria. An optimal protocol for the prevention of neonatal sepsis has not yet been developed. While intrapartum antimicrobial prophylaxis appears to provide the best potential, each of the currently suggested protocols has significant drawbacks. Drawbacks include the potential for missing high-risk carriers, failure to treat a significant proportion of those destined to deliver an affected infant because no risk factors are present, and empirical treatment of a large proportion of the population in order to present significant disease in a few. Until an effective program of immunization becomes available, intrapartum prophylaxis of group B streptococcal carriers appears to offer the best hope of reducing the incidence of neonatal disease. Caregivers should adopt a uniform practice with regard to screening and prophylaxis. It is essential that any broad-based screening program include an evaluation of efficacy as well as complications including the development of new etiologic agents as causes of neonatal sepsis and the emergence of resistant bacteria. Further, mothers and newborns should be evaluated for drug adverse reactions and the impact of intrapartum prophylaxis on the use of prolonged empirical broad-spectrum antimicrobial therapy on the asymptomatic infant. Additional research is necessary regarding the required duration of therapy for optimal effect of intrapartum prophylaxis, the need for postnatal prophylaxis of the asymptomatic neonate, and the optimal agent for neonatal prophylaxis (penicillin versus broad-spectrum agents) if neonatal therapy is necessary after intrapartum prophylaxis.

Atlantoaxial instability in Down's syndrome: a case report and review.

Down's syndrome is the most common autosomal chromosomal abnormality in humans and is associated with a number of well known clinical findings. Atlantoaxial instability is a less recognized, yet potentially significant, manifestation of Down's that has gained importance because of the widespread participation of Down's syndrome individuals in athletic events. Early recognition and appropriate management of patients with atlantoaxial instability can significantly reduce the morbidity and mortality associated with this condition and guide patients and their parents toward continued safe participation in athletics.

Preparation, metabolic stability and biological properties of omega-trifluorinated analog of 12-hydroxyeicosatetraenoic acid.

12-Hydroxyeicosatetraenoic acid (HETE) is associated with a variety of inflammatory conditions. For studies on pathophysiological function of 12-HETE, metabolically more stable analogs of 12-HETE would be useful. We biologically synthesized 20,20,20-trifluoro-12-HETE (20-F3-12-HETE) by incubating enantioselectively synthesized 20,20,20-trifluoro-arachidonic acid with human platelets. The product was identified by UV absorption spectrophotometry and gas chromatography-mass spectrometry. When 1 microgram 20-F3-12-HETE was incubated with 5 X 10(6) human neutrophils for 45 min, only 5% of the analog was metabolized while 66% of 12-HETE was metabolized in the same incubation condition. With 2 X 10(7) neutrophils, 37% of the analog was metabolized at the same incubation condition while 87% of 12-HETE was metabolized. Thus, by blocking omega-oxidation of 12-HETE with fluorine atoms, the stability of 12-HETE was greatly increased. This result indicates that the omega-oxidation is a major pathway for 12-HETE metabolism. The analog demonstrated as much chemotactic activity on human neutrophils as 12-HETE, and binding affinity of the analog for 12-HETE receptor in human epidermal cell was equal to that of 12-HETE. An analog of 12-HETE, which has extended metabolic stability without alteration of neutrophil chemotactic activity and binding affinity, would be a useful tool for studies on pathophysiological role of 12-HETE in inflammatory conditions.

Angular deformities of the tibia in children: a review of three cases.

Children born with prominent bowing of the tibia are an obvious concern to parents and physicians. Managing these congenital angulations requires early and accurate diagnosis. Classification is based on the direction of the angulation and its associated pathology. The three basic classes of tibial angulations commonly accepted are based on the work of Heyman and Herndon. The success of treating these children depends on the type and severity of the angulation. This article presents three cases of tibial bowing and describes the treatment and prognosis of each.

The management of intoeing: a review.

Children presenting with an intoeing gait are the most common pediatric problem seen by orthopedists. Most parents are concerned that this rotational problem may result in a permanent disability or impediment of their child's physical performance. The etiologies of these conditions are still debated, although the structural conditions giving rise to intoeing can be correlated with the age of onset. The extent of the rotational problem is determined through physical examination and the measurement of indicative angles. A rotational profile is compiled from the values obtained, allowing the child's progress to be followed. Many children spontaneously resolve the structural problems responsible for their intoeing gait. The success one attributed to the traditional methods for treating intoeing is now believed to be the result of this natural resolution.

Biosynthesis of 20,20,20-trifluoroleukotriene B4 from 20,20,20-trifluoroarachidonic acid: a metabolically stable analog of leukotriene B4 and its application to a study of stimulation of leukotriene B4 synthesis by immunoglobulin G1,2.

Leukotriene B4 is rapidly metabolized through omega-oxidation, preventing its detection when it is produced under certain biological conditions. To investigate leukotriene B4 production in various physiological conditions, analogs of arachidonic acid which are converted to metabolically stable analogs of leukotriene B4 would be useful. We have synthesized 20,20,20-trifluoroarachidonic acid by the cis-selective Wittig reaction of the C12-C20 fragment with phosphonium salt. 20,20,20-trifluoroarachidonic acid was transformed into 20,20,20-trifluoroleukotriene B4 when incubated with human neutrophils in the presence of the calcium ionophore A23187. The product was identified by uv absorption spectrophotometry, gas chromatography-mass spectrometry, and coelution on high-performance liquid chromatography with 20,20,20-trifluoroleukotriene B4, which was enantioselectively synthesized by the reaction of the fluorine-containing C11-C20 fragment with the C1-C10 phosphonate. The fluorinated leukotriene B4 demonstrated as much chemotactic activity on human neutrophils as natural leukotriene B4 and was metabolically stable when incubated with human neutrophils, probably by blocking omega-oxidation. Also, enzymes catalyzing the transformation of arachidonic acid (AA) into leukotriene B4 did not discriminate the fluorinated precursors from the natural, nonfluorinated AA, thus 20-F3-AA is a valid analog of AA to be used in the study of AA metabolism. When 50 microM of the fluorinated acid was incubated with neutrophils stimulated with heat-aggregated human immunoglobulin G, a significant amount of fluorinated leukotriene B4 (4.3 ng/10(6) cells/40 min, at most) was formed in a dose-dependent manner while little leukotriene B4 was detected with incubation with 50 microM arachidonic acid, probably due to omega-oxidation of the product, leukotriene B4. 20,20,20-Trifluoroarachidonic acid appears to be a useful tool for studying the capacity of leukotriene B4 synthesis in various biological systems while long-lasting 20,20,20-trifluoroleukotriene B4 would serve as an excellent analog of leukotriene B4 in pharmacological studies to understand functions of leukotrienes B4.

Comparative neurotoxicity and pyrrole-forming potential of 2,5-hexanedione and perdeuterio-2,5-hexanedione in the rat.

2,5-Hexanedione (2,5-HD), the neurotoxic metabolite of n-hexane, reacts with protein amines to form alkylpyrrole adducts. Pyrrolylation of neurofilament protein may be the initiating molecular event in 2,5-HD neuropathy. The present study compares the neurotoxic and pyrrole-forming potentials of 2,5-HD with those of perdeuterio-2,5-HD ([D10]-2,5-HD) in the rat. Due to a requirement for C-H bond breaking in the reaction mechanism, the latter derivative was expected to exhibit a primary isotope effect, thus forming the pyrrole at a slower rate. In vitro studies confirmed that [D10]-2,5-HD pyrrolylated protein at only one-third of the initial rate seen with native 2,5-HD. Prolonged incubation resulted in similar pyrrole concentrations with both derivatives. Adult, male Wistar rats were administered daily (5 days/week) ip doses of either 3.5 mmol 2,5-HD or [D10]-2,5-HD/kg/day for 17 days or 2.5 mmol/kg/day for 38 days. At termination, animals administered 2,5-HD and [D10]-2,5-HD exhibited 27 and 8% body weight loss, respectively. Moderate to severe hindlimb paralysis was present in the 2,5-HD groups while only mild effects were seen in [D10]-2,5-HD-dosed rats. Neuropathological changes were prominent in spinal cord sections from 2,5-HD-treated animals, while no effects were present in rats given the deuterated derivative. Pyrrole adduct concentrations in serum and axonal cytoskeletal proteins from 2,5-HD-treated animals were two- to threefold higher than in rats given equimolar doses of [D10]-2,5-HD. Levels of covalent crosslinking of axonal cytoskeletal proteins (assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) appeared to correlate with pyrrole concentrations. Tissue concentrations of each diketone isomer were not significantly different, indicating similar uptake of native and deuterated 2,5-HD. Mass spectrometry revealed rapid back exchange of the terminal (methyl) but not of the internal (methylene) deuteriums of [D10]-2,5-HD in vivo. These findings support an absolute requirement for pyrrole formation in gamma-diketone neurotoxicity.

Identification of vitamin K1 chromenol--a novel metabolite of vitamin K1 formed in vitro by a component in blood.

A newly recognized metabolite of vitamin K1, vitamin K1 chromenol, is produced when the vitamin is added to the plasma or serum of a number of species. The metabolite was identified by comparison of its uv and mass spectra and high-performance liquid chromatographic retention times with those of the synthetic vitamin K1 chromenol. In aqueous solution vitamin K chromenol decomposed to a variety of products and reacted with nucleophilic substances. Optimal conditions for its formation and evidence that chromenol formation may be an enzyme catalyzed reaction are presented.

The effects of Love Canal soil extracts on maternal health and fetal development in rats.

The effects of a solvent extract of the surface soil of the Love Canal chemical dump site, Niagara Falls, New York, and of a natural extract, or leachate, which is drained from the canal for treatment, on the maternal health and fetal development were determined in rats. The solvent extract, which was contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (2, 3,7,8-TCDD) at 170 ppb and numerous other chlorinated organic compounds with the primary identified components being the isomers of benzenehexachloride (BHC), was dissolved in corn oil and administered by gavage to pregnant rats at 0,25,75, or 150 mg crude extract/kg/day on Days 6-15 of gestation. A 67% mortality was observed at the highest dose. The rats were sacrificed on Day 20. Dose-related increases in relative liver weight accompanied by hepatocyte hypertrophy were observed at all dose levels. Fetal birthweight was decreased at 75 and 150 mg extract/kg/day. No major treatment-related soft tissue or skeletal malformations, except for delayed ossification, were observed. Based on literature values for BHC, all of the observed toxicity could be accounted for by the BHC contaminants of the extract. The crude organic phase of the leachate was administered to pregnant rats at 0,10,100, or 250 mg/kg/day as described above. Maternal weight gain decreased at 100 and 250 mg/kg/day, accompanied by 5 and 14% maternal mortality, and 1 and 3 dead fetuses, respectively. Early resorptions and the percentage of dead implants increased whereas fetal birthweights were decreased at 250 mg/kg/day. No major treatment-related soft tissue or skeletal malformations, except for delayed ossification, were observed. The primary components of the complex leachate by mass were tetrachloroethanes; however, 2,3,7,8-TCDD, which was present at 3 ppm, probably accounted for all the observed toxicity.

Differences in the chromatographic and mass spectral properties of 3,3',5,5'-tetrachlorodiphenoquinone and 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Tetrachlorodiphenoquinones have the same exact mass and elemental composition as the toxic environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin. However, analysis of 3,3'-5,5'-tetrachlorodiphenoquinone showed a pronounced tendency toward chemical reduction in the mass spectrometer to the quinol compound, producing a molecular ion two mass units higher than 2,3,7,8-tetrachlorodibenzo-p-dioxin. Distinct differences were also apparent between the mass spectral fragmentation patterns of 3,3',5,5'-tetrachlorodiphenoquinone and 2,3,7,8-tetrachloridibenzo-p-dioxin. The 3,3',5,5'-tetrachlorodiphenoquinone spectrum shows a successive loss of carbon monoxide, with the most prominent fragment corresponding to loss of two molecules of carbon monoxide plus chlorine. In the mass fragmentation of 2,3,7,8-tetrachlorodibenzo-p-dioxin carbon monoxide loss is suppressed, but loss of one molecule of carbon monoxide plus chlorine is a major fragment ion. During an alumina column clean-up procedure 3,3',5,5'-tetrachlorodiphenoquinone did not coelute with the fraction containing 2,3,7,8-tetrachlorodibenzo-p-dioxin. This evidence indicates that tetrachlorodiphenoquinones are unlikely to interfere with mass spectrometric determination of 2,3,7,8-tetrachlorodibenzo-p-dioxin in environmental samples.

Subchronic exposure of mice to Love Canal soil contaminants.

The health hazard potential of soil collected from the surface of the Love Canal chemical dump site in Niagara Falls, New York, was assessed in 90-day exposure studies. Female CD-1 mice were exposed to two concentrations of the volatile components of 1 kg of soil with and without direct soil contact. Control mice were identically housed but without soil. The soil was replaced weekly and 87 compounds were detected in the air in the cages above fresh and 7-day-old soil as analyzed by gas chromatography/mass spectrometry. The concentration of many of these compounds decreased during the 7-day exposure cycle. Histopathologic, hematologic, and serum enzyme studies followed necropsy of all mice. There was no mortality of mice exposed for up to 90 days under any condition. Thymus and spleen weights relative to body weight were increased after 4 weeks of exposure by inhalation but not after 8 or 12 weeks of exposure. alpha-, beta-, and delta- Benzenehexachlorides , pentachlorobenzene, and hexachlorobenzene were detected in liver tissue from these animals. Mice exposed to 5- to 10-fold elevated concentration of volatiles had increased body and relative kidney weights. There was no chemically induced lesion in any animal exposed only to the volatile soil contaminants. Mice exposed by direct contact with the soil without elevated volatile exposure had increased body (10%) and relative liver weights (169%). Centrolobular hepatocyte hypertrophy, which involved 40 to 70% of the lobules, was observed in all mice in this group.(ABSTRACT TRUNCATED AT 250 WORDS)

Increased responsiveness of the hepatic guanylate cyclase-guanosine 3',5'-monophosphate system to nitrosoguanidine following partial hepatectomy.

When tested at concentrations producing submaximal responses, the N-nitroso carcinogen, N-methyl-N'-nitro-N-nitrosoguanidine (methylnitro-nitrosoguanidine) elicited a 2-fold greater increase in guanosine 3',5'-monophosphate (cyclic GMP) accumulation in slices and a 5-fold greater stimulation of guanylate cyclase activity in whole homogenates of rat liver examined 24 h after 75% hepatectomy compared to the corresponding methylnitro-nitrosoguanidine responses in sham-operated and unoperated controls. Enhanced methylnitro-nitrosoguanidine sensitivity of guanylate cyclase in whole homogenates of regenerating liver was attributable to altered responsiveness of the enzyme activity of the 100 000 X g soluble fraction, which contained 98% of the methylnitro-nitrosoguanidine responsive activity. Basal cyclic GMP accumulation and guanylate cyclase activities of these systems, and their responses to concentrations of methylnitro-nitrosoguanidine eliciting maximal stimulation were unchanged after partial hepatectomy or sham operation, compared to unoperated controls. The findings of (a) increased heme concentrations in the supernatant and the high molecular weight Sephadex G-25 fraction of sham operated, compared to regenerating liver, (b) suppression of methylnitro-nitrosoguanidine responsive activity after addition of exogenous hemoglobin to supernatants from regenerating liver, and (c) enhancement of the responsiveness of soluble guanylate cyclase from sham operated liver to submaximal methylnitro-nitrosoguanidine after reduction of endogenous heme content by in situ perfusion, all suggested that the difference in methylnitro-nitrosoguanidine action observed in control vs. regenerating liver are related to a lower heme-protein content of the latter. These results emphasize the importance of endogenous heme as a factor modulating the response of the hepatic guanylate cyclase system to methylnitro-nitrosoguanidine.

Calcium-dependent modulation of guanosine 3',5'-monophosphate in renal cortex. Possible relationship to calcium-dependent release of fatty acid.

The effects of Ca(2+) on cGMP accumulation in rat renal cortical slices were correlated with the effects on (14)C-fatty acid release in tissue prelabeled with (14)C arachidonate. Ca(2+) in the presence and absence of ionophore A23187 exerted parallel effects on the release of labeled arachidonate from slices and on slice cGMP content. Thus, Ca(2+) stimulated both arachidonate release and tissue cGMP accumulation 2 to 3-fold when added to slices of renal cortex previously deprived of Ca(2+) and Mg(2+), whereas Mg(2+) had no stimulatory effect on either arachidonate release or tissue cGMP content. In the presence of A23187, Ca(2+) increased arachidonate release and tissue cGMP accumulation 4 to 6-fold. Tetracaine partially inhibited Ca(2+)-induced arachidonate release and completely blocked Ca(2+)-induced cGMP accumulation. Ca(2+)-induced arachidonate release was unaffected by the absence of O(2). Addition of exogenous arachidonate to slices of renal cortex increased tissue cGMP content 2-fold. Linoleate exerted a lesser effect on tissue cGMP, while palmitate and oleate had no effect. Ca(2+)- and arachidonateinduced cGMP contents in renal cortical slices were not additive, and both were abolished by exclusion of O(2). Since nitroprusside increased cGMP accumulation 10- to 15-fold in O(2)-deprived slices, loss of the Ca(2+) and arachidonate responses under these incubation conditions was selective. Ca(2+)-induced cGMP accumulation was unaffected by indomethacin (100 microg/ml), but was abolished by 200 microM 5,8,11,14-eicosatetraynoic acid (TYA). The results are consistent with the possibility that the Ca(2+)-dependent processes regulating cGMP in renal cortex include Ca(2+)-dependent acyl hydrolase activity, which limits the availability of free polyunsaturated fatty acids. A role for fatty acid oxygenation products in the stimulation of cGMP is suggested, but not established, by the O(2) dependence of the actions of both Ca(2+) and exogenous fatty acids. The failure of exogenous arachidonate or linoleate to mimic quantitatively the actions of Ca(2+) on cGMP may reflect the involvement of other Ca(2+)- and O(2)-dependent processes in modulation of cGMP in this tissue or limited access of exogenous fatty acid to cGMP regulatory sites in the cell.

Further characterization of human erythrocyte superoxide dismutase.

1. A simplified procedure for the preparation of highly purified human superoxide dismutase from erythrocytes was developed which avoided extremes of pH and ionic strength and the use of organic solvents; the properties of human and bovine proteins, prepared by the method, were compared. 2. Using the two dimensional electrophoretic procedure of O'Farrell, the human superoxide dismutase was found to consist of a single type of polypeptide. 3. The human protein was found to have a total of eight half-cystine residues per mole of protein, compared to six such residues for the bovine protein. The human protein has two sulfhydryl groups which are reactive toward mercurials when dissolved in 1M guanidine-hydrochloride and approximately 3 reactive sulfhydrls when the protein is dissolved in 6 M guanidine hydrochloride. The distribution of the eight sulfur atoms appears to consist of four involved in disulfide linkages, two deeply buried within the molecule and unreactive except under strongly denaturing conditions, and two which are reactive under mildly denaturing conditions. No zero-valent sulfur was found. 4. The visible optical absorption, the visible circular dichroism, and the electron paramagnetic resonance spectra are essentially identical with those of the bovine protein. No unusual absorbance was found at 330 nm. The near ultraviolet spectrum is different from that of the bovine protein, and this appears to be due to differing amino acid compositions. 5. Two fractions of superoxide dismutase activity were observed during chromatography of partially purified solutions on diethylaminoethyl-cellulose. The minor, less mobile form, was found to revert to the less mobile species on aging; the reverse process was not observed to occur. The minor component was found to contain equimolar amounts of Zn and Cu and to have a specific dismutase activity somewhat higher than that of the purified major fraction.

Sulfhydryl reactivity of human erythrocyte superoxide dismutase. On the origin of the unusual spectral properties of the protein when prepared by a procedure utilizing chloroform and ethanol for the precipitation of hemoglobin.

1. During purification of human superoxide dismutase by the McCord-Fridovich procedure (McCord, J.M. and Fridovich, I. (1969) J. Biol. Chem. 244, 6049--6055) the "extra" sulfhydryl groups react with a variety of sulfur containing compounds including zero-valent sulfur to yield several dismutase fractions containing excess sulfur atoms and having a unique absorption band in the region of 325 nm. This is shown to be artefact of the purification procedure. 2. Cysteine trisulfide and glutathione polysulfide were found to react with native human superoxide dismutase to yield derivatives having no reactive sulfhydryl groups and possessing spectral properties similar to the various fractions obtainable from the above purification procedure. A structure of the type protein-CH2-S-(S)n R is proposed to account for the results. The value of n is variable, and the additional sulfur reactive toward thiol reagents is thought to be due to persulfides (R = H). The 325 nm band is probably due to a n leads to sigma ss transition associated with a strained S-S bound.

Lipid accumulation cells derived from porcine aorta and grown under anaerobic conditions.

Fibroblast-like cells, derived from porcine aorta, were cultured under aerobic and anaerobic conditions. Light and electron microscopic examinations, lipid composition measurements, and incorporation of radioactive precursors into lipids of these cells were performed. Anaerobically grown cells accumulated oil red O stainable droplets and within 6 hr the triacylglycerol content increased to 4 times the level determined in cells grown under aerobic conditions. This ratio remained constant throughout an additional 12 hr of growth. The fatty acid composition of the triacylglycerols which accumulated under anaerobic conditions differed from the composition of fatty acids in the triacylglycerols present in the growth medium. The cellular unesterified fatty acids of the anaerobically grown cells differed only slightly in composition from the fatty acids in the growth medium, while the unesterfied fatty acids of aerobically grown cells differed to a greater extent from those of the growth medium.

Studies on the reconstitution of bovine erythrocyte superoxide dismutase. V. Preparation and properties of derivatives in which both zinc and copper sites contain copper.

1. We have developed a procedure for preparing derivatives of bovine superoxide dismutase in which primarily the Cu binding sites are occupied by Cu2+ (2 Cu2+-) and in which both the Zn and Cu binding sites are occupied by Cu2+ (4 Cu2+-). 2. The 2 Cu2+ protein shows approximately one-half the superoxide dismutase activity of an equivalent amount of native protein. A two-fold enhancement of the activity of 2 Cu2+-dismutase was observed upon occupation of the Zn sites either with Zn2+ or Cu2+. 3. The electron paramagnetic resonance spectrum of 4 Cu2+ protein was recorded over the temperature range 5-100 degrees K and the results suggest an antiferro-magnetic interaction between Cu2+ in the Zn site and Cu2+ in the Cu site having a coupling constant of approx. 52 cm-1. 4. The binuclear Cu2+ complex was found to accept only one electron from ferrocyanide. 5. One-half the total Cu+ of dithionite reduced 4 Cu+ protein was found to react rapidly with bathocupreine sulfonate whereas the other half reacted slowly. Reduced native protein did not react with bathocupreine sulfonate below 70 degrees C.