Focus: Sex and Gender Health: Tobacco Smoking and the Resting Maternal Brain: A Preliminary Study of Frontal EEG

Tobacco smoking has been attributed to a wide range of detrimental health consequences for both women and their children. In addition to its known physical health effects, smoking may also impact maternal neural responses and subsequent caregiving behavior. To begin investigating this issue, we employed electroencephalography (EEG) to examine resting neural oscillations of tobacco-smoking mothers (n = 35) and non-smoking mothers (n = 35). We examined seven EEG frequency bands recorded from frontal electrode sites (delta, theta, alpha, alpha₁, alpha₂, beta, and gamma). While no between-group differences were present in high-frequency bands (alpha₂, beta, gamma), smokers showed greater spectral power in low-frequency bands (delta, theta, alpha, alpha₁) compared to non-smokers. This increased power in low-frequency bands of tobacco-smoking mothers is consistent with a less aroused state and may be one mechanism through which smoking might affect the maternal brain and caregiving behavior.     

Immunological studies of the anticryptococcal factor of normal human serum

Preliminary studies have shown a very high inhibitory activity in the alpha₂ and gamma zone of human serum towards the growth of Cryptococcus neoformans. These findings are now corroborated by single radial immunodiffusion tests, which showed the some loss of IgA and IgM globulins and of the other three globulin fractions (ceruloplasmin, alpha₂ macroglobulin and alpha₂ HS glycoprotein) which migrate in the alpha₂ zone. The data was obtained by single radial immunodiffusion tests. The losses were not statistically significant however. No change in the immunoglobulin content of the sera kept for 6 days in contact with a heat-killed suspension of C. neoformans was noted. These findings suggest, that the inhibitory activity of the normal human serum on the in-vitro growth of C. neoformans is due to the above mentioned globulin fractions and not to a single specific factor.     

Human alpha-chain globin messenger: prediction of a nucleotide sequence

The presence in human serum of inhibitory activity to rat liver insulin specific protease has been detected in an alpha₁ globulin preparation (Cohn Fraction IV₁). Separation into four components and partial purification (40 to 107 fold) has been achieved by heat denaturation of non-active protein, Sephadex G-100 gel filtration and ion-exchange chromatography upon QAE Sephadex. Each of the inhibitors was found to be competitive in nature. The molecular weight of the inhibitors is between 4,000–7,000 and the activity is destroyed for the most part by chymotrypsin.     

A serum protein affecting the regulation of the immune system

A thermostable alpha₂ globulin inhibiting the immunoglobulin/anti-immunoglobulin reaction was demonstrated working with subacute sclerosing panencephalitis (SSPE) and control serum IgG. This protein was isolated from SSPE and normal human blood, it inhibits the immunoglobulin/anti-immunoglobulin reaction but no other antigen/antibody reactions when applying different immunochemical methods such as nitrocellulose immunofixation, 2 site immunoradiometric assay, solid phase radioimmunoassay in coated cups. This was demonstrated, working on the one hand with measles virus strain Edmonston or SSPE virus strain D.R. and SSPE serum and on the other hand with IgG from SSPE and control serum. This alpha₂ globulin, an inhibiting protein, appears to be related to normal immunosuppressive protein.Special Issue dedicated to Dr. Elizabeth Roboz-Einstein.     

The human corticosteroid binding globulin gene is located on chromosome 14q31–q32.1 near two other serine protease inhibitor genes

Summary Human corticosteroid binding globulin (CBG) cDNA fragments were radiolabeled and hybridized in situ to metaphase chromosome preparations. The results localized the CBG gene to the q31–q32.1 region of human chromosome 14. This location also contains the genes for two closely related serine protease inhibitors: alpha₁-proteinase inhibitor and alpha₁-antichymotrypsin. It is therefore likely that these genes evolved by duplication events, and it would appear that this region contains a series of functionally related genes.     

Inhibition by serum of encephalitogenic activity of myelin basic protein: nature of the serum factor responsible.

The capacity of basic protein of myelin (BPM) from various species to produce experimental autoimmune encephalomyelitis (EAE) in the guinea pig or the Lewis rat, was abrogated by mixing BPM with alpha₂ macroglobulin, transferrin, and ceruloplasmin. Inhibition of EAE was observed only when the encephalitogen was held in vitro directly with the inhibitor and not when each was injected separately. Guinea pigs which failed to develop EAE after an injection of a mixture of BPM with alpha₂ macroglobulin nevertheless developed cell-mediated immunity and antibodies to BPM. However, animals injected with a mixture of the encephalitogenic tryptophan peptide of BPM with alpha₂ macroglobulin developed neither cell-mediated immunity nor humoral antibody to BPM. Inhibition of EAE in the present studies was attributed to binding between the inhibitor and BPM, which possibly blocked its main encephalitogenic determinant, rather than enzymatic cleavage of BPM. This inhibitory effect of alpha globulin on immunogenicity could represent a further example of homeostatic control over immune responses.     

Gc,Tf, Pi,PLG, AHS and F13B in two Czech populations

Six serum protein systems, Gc globulin (Gc), transferrin (Tf), alpha₁-antitrypsin (Pi), plasminogen (PLG), alpha2-HS-glycoprotein (AHS), and coaglulation factor XIII B (F13B) have been studied in two Czech populations from Prague (n=243) and Olomouc (n=205). The results are discussed with reference to other investigations in Middle Europe.     

Differential regulation of integrin-mediated proplatelet formation and megakaryocyte spreading

Guinea pig bone marrow megakaryocytes were cultured on a type I rat tail collagen gel which stimulated proplatelet formation. Proplatelet formation was inhibited by monoclonal antibody LM609 to the alpha_v beta₃ integrin (VnR), but not by monoclonal antibodies to the alpha₅, alpha₆, beta₁, or IIb beta_{3(GPIIb–IIIa)} integrin proteins. Megakaryocytes cultured on a plastic surface and stimulated with thrombin undergo a spreading and an adhesion reaction. This reaction is blocked in a dose-dependent manner by the tetrapeptide RGDS and by the monoclonal antibody PG2 to the GPIIb–IIIa integrin, but not by the monoclonal antibody LM609 to the VnR. Immunoprecipitation and affinity chromatography experiments demonstrate that guinea pig megakaryocytes have distinct GPIIb–IIIa and VnR integrins with similar electrophoretic mobility. Spreading was significantly inhibited in a dose-dependent fashion by drugs which elevate cellular cyclic AMP, including forskolin, dibutyryl cAMP, and isobutylmethylxanthine. In contrast to spreading, megakaryocyte proplatelet formation was stimulated by these agents in a dose-dependent manner. Megakaryocyte spreading was stimulated by the protein kinase C (PKC) activator phorbol myristate acetate (PMA) and inhibited by the PKC inhibitors Calphostin C and K5720 in a dose-dependent manner. PKC inhibitors did not inhibit megakaryocyte proplatelet formation. These results demonstrate that the closely related VnR and GPIIb-IIIa integrins regulate different aspects of megakaryocyte morphological change and appear to be associated with different second messenger systems. © 1995 Wiley-Liss, Inc.     

Alpha-adrenergic receptors in liver membranes: delineation with subtype selective radioligands

Alpha₁ and alpha₂ adrenergic receptors have previously been demonstrated in rat liver membranes by competition curves of [³H]dihydroergocryptine ([³H]DHE) with the alpha₁ selective antagonist prazosin (B.B. Hoffman, D. Mullikin-Kilpatrick and R.J. Lefkowitz, J. Biol. Chem. $\text{255}$:4645–4652, 1980). The present studies have utilized the radioligands [³H]prazosin and [³H]yohimbine to further define alpha receptors in rat liver membranes. [³H]Prazosin was found to label alpha₁ receptors whereas [³H]yohimbine labelled alpha₂ receptors. The proportion of alpha₁ and alpha₂ receptors determined directly with these radioligands (79% and 20% respectively) was in good agreement with the proportions determined previously with [³H]DHE. Guanine nucleotides were found to reduce the affinity of the agonist epinephrine at the alpha₂ sites labelled by [³H]yohimbine but not at the alpha₁ sites labelled by [³H]prazosin. These results have implications for the interpretation of agonist interactions with alpha receptors in liver membranes.     

Alpha-adrenergic receptor subtypes: quantitative assessment by ligand binding.

We describe a method for quantitatively determining the alpha- adrenergic receptor subtypes in membrane fractions by studying the displacement of [³H] dihydroergocryptine by selective alpha antagonists and analyzing this data by a computer modeling technique. Alpha₁ receptors are those with a higher affinity for prazosin than for yohimbine; alpha₂ receptors have a higher affinity for yohimbine than for prazosin. Phentolamine does not discriminate between the two receptor subtypes present in rabbit uterus. The alpha receptor population of rabbit uterus was found to be 37% alpha₁ receptors and 63% alpha₂ receptors. The human platelet and rat liver alpha receptors were determined to be exclusively alpha₂ and alpha₁, respectively. In the uterus, prazosin had a 8800 fold greater affinity for alpha₁ than alpha₂ receptors while yohimbine had a 510 fold greater affinity for alpha₂ than alpha₁ receptors. The use of [³H] dihydroergocryptine displacement curves generated with selective alpha receptor antagonists coupled with subsequent computer modeling provides a precise and powerful method for quantifying the alpha receptor population of a tissue; this technique should be of value in studying the detailed regulation of alpha receptors in tissues which have both alpha₁ and alpha₂ receptors.     

The Electrophoretical Determination of Serum Protein Fractions in Lycopene Treated Experimental Diabetic Rats

This study was planned to determine the effects of lycopene treatment on serum protein fractions in experimental diabetic rats. In order to induce diabetes in rats in the diabetes (D) and diabetes + lycopene (DL) groups, rats were given 45 mg/kg single-dose streptozotocin intraperitoneally. Lycopene (10 mg/kg/day dissolved in sunflower oil) was administered to the rats in the lycopene-only (L) and DL groups. Blood glucose levels and HbA1c% in DL group and diabetes group increased (p < 0.05) compared to control and L group. Total protein, albumin, α₁, α₂, and β globulin fractions of diabetic and DL groups were lower than control and L groups (p < 0.05). D group had lowest gamma (γ) globulin levels among other groups (p < 0.05). The γ globulin levels was slightly increased than diabetic groups (D and DL), but it was still lower than control and L groups (p < 0.05). The highest value of A/G ratio was observed in diabetic group. Similarly, the % level of A/G ratio of D group was higher than other groups. It was noted that the A/G ratio decreased and reached to control group levels after lycopene treatment.     

Evidence for a novel circulating alpha 1 protein in tumour-bearing rats. Differentiation from acute phase alpha 1 proteins and from alpha 1 fetoprotein

It has been found in this study that the serum from rats bearing a transplanted dibenzanthracene-induced tumour ($\text{RD}\text{3}$), has a high concentration of alpha₁ proteins compared with normal rat serum. These alpha₁ proteins have been isolated by an immunoabsorption method and have been compared by immunological methods with the acute phase alpha₁ proteins isolated by the same method from the serum of rats presenting an inflammatory reaction. It has been found that the isolated $\text{RD}\text{3}$ alpha₁ proteins were composed of two major proteins: one of these corresponded to an inflammatory protein, the alpha₁-AP-globulin. The other may be a new protein, as it is absent from the serum of rats with an acute phase inflammatory reaction and nor does it correspond to alpha₁ feto-protein, a carcino-embryonic protein presenting the same electrophoretic mobility.     

Contribution to the structural characterization of gamma globulin from pig colostrum

From the results obtained in the present work it is concluded that gamma globulin of the 7 S type (γG) which represents the main immunoglobulin component of pig colostrum, differs from serum γG globulin by the presence of another type of polypeptide chain, designed L₂; the latter was detected after S-sulfonation in starch gel electrophoresis as the fastest moving component and its immunoelectrophoretic pattern shows the presence of two precipitating components. Preparation of soluble heavy and light chains enabled us to study them imunochemically. The heavy chain is represented by two zones; the fainter one was not detected in comparative analysis of the heavy chain of serum γG globulin. The L₁ chain of colostral gamma globulin and the light chain of serum gamma globulin seem to contain three precipitating components, one of which appears due to aging of the chain solution in the presence of glycine. The material from colostrum seems to contain a larger amount of this component than serum. Further it was shown that the component arising in this way is antigenically closely related to the heavy chain.     

Biosynthesis of bacterial glycogen: purification and characterization of ADPglucose pyrophosphorylase with modified regulatory properties from Escherichia coli B mutant CL1136-504

The l-thyroxine binding site in human serum thyroxine-binding globulin was investigated by affinity labeling with N-bromoacetyl-l-thyroxine (BrAcT₄). Competitive binding studies showed that, in the presence of 100 molar excess of BrAcT₄, binding of thyroxine to thyroxine-binding globulin was nearly totally abolished. The reaction of BrAcT₄ to form covalent binding was inhibited in the presence of thyroxine and the affinity-labeled thyroxinebinding globulin lost its ability to bind thyroxine. These results indicate BrAcT₄ and thyroxine competed for the same binding site. Affinity labeling with 2 mol of BrAcT₄/mol of thyroxine-binding globulin resulted in the covalent attachment of 0.7 mol of ligand. By amino acid analysis and high voltage paper electrophoresis, methionine was identified as the major residue labeled (75%). Lysine, tyrosine, and histidine were also found to be labeled to the extent of 8, 8, and 5%, respectively.     

Neurochemical evidence for two types of presynaptic alpha2-adrenoceptors

Neurochemical and pharmacological evidence has been obtained that noradrenergic varicosities (in mouse and rat vas deferens) and cholinergic varicosities (in the Auerbach's plexus) contain heterogenous alpha₂-adrenoceptors through which the release of [³H]noradrenaline and [³H]acetylcholine can be modulated. The quantitative data also support the hypothesis that different noradrenaline and xylazine sensitive alpha₂-adrenoceptors are present prejunctionally in the vas deferens and Auerbach's plexus preparations. Prazosin, although it has a presynaptic inhibitory effect on alpha₂-adrenoceptors of noradrenergic axon terminals, has no effect on cholinergic axon terminals. These data suggest that there are two different types of alpha₂-adrenoceptors at the presynaptic axon terminals.Special Issue Dedicated to Dr. Abel Lajtha     

Possible interaction of PGF2 alpha with hypothalamus-pituitary-thyroid axis in man

The possible interactions of PGF_{2 alpha} on the hypothalamus-pituitary-thyroid axis are the object of this study.Firstly a significant direct effect of PGF_{2 alpha} infusion (mg2, 5/270 min) on TSH,PRL,LH,FSH and GH pituitary secretion was excluded.Thereafter the possible PGF_{2 alpha} on PRL and TSH pituitary response to TRH was considered: in only two cases PGF_{2 alpha} was able to increase the TSH response.Finally the Authors studied T₃ response to endogenous TSH rise induced by TRH: if they consider the mean peak responses of T₃ the increase is significant only when PGF_{2 alpha} infusion is performed.     

Impact of Ty3/<Emphasis Type="Italic">Gypsy</Emphasis> group retrotransposon <Emphasis Type="Italic">Lila</Emphasis> on the D-Genome specificity of common wheat <Emphasis Type="Italic">Triticum aestivum</Emphasis> L.

An analysis of the primary structure of BAC clone 112D20 T. aestivum, that contains D-genome specific Ty3-Gypsy-retrotransposon Lila is presented. PCR analysis of nulli-tetrasomic and deletion lines of T. aestivum allowed to localize this BAC clone in the distal region of the long arm of chromosome 5D. Characteristic feature of BAC clone 112D20 is a high concentration of Ty3-Gypsy-retrotransposons (61.7%), and low content of the genes (1.2%). Only a single open reading frame was revealed homologous to an unknown gene of Ae. tauschii. Specific to the D-genome Ty3-Gypsy-retrotransposon Lila in the BAC clone 112D20 is 14 kb in length and contains unequal in size long terminal repeats. The data of in situ hybridization and PCR analysis of different Triticeae species suggest that this retroelement was amplified within the ancestral species of Ae. tauschii, the donor D-genome. The suggested time of amplification based on estimation of insertion time of Lila 112D20 is 1.7 million years, which corresponds to the formation of the first allopolyploid forms of wheat. Based on comparison with the previously obtained data, it is concluded that the amplification of retroelements specific to each genome of wheat took place during formation of the diploid progenitors of these genomes.     

Phorbol esters inhibit alpha1 adrenergic stimulation of glycogenolysis in isolated rat hepatocytes

Tumor promoting phorbol esters can stimulate Ca⁺⁺-phospholipid-dependent protein kinase. It has been suggested that this enzyme may mediate the effects of calcium-dependent hormones. In this paper the effects of phorbol 12-myristate 13-acetate (TPA) on isolated rat hepatocyte metabolism were studied. Phorbol esters completely blocked alpha₁-adrenergic stimulation of glycogenolysis. This effect is quite specific for alpha₁-adrenergic actions, as the stimulations of glycogenolysis by vasopressin, angiotensin II, ionophore A-23187 and glucagon were unaffected by TPA. The potencies of the different phorbol esters used in this study suggests that the inhibitory effects of these agents may be due to activation of protein kinase C. The effect of phorbol esters on alpha₁-adrenergic actions seems to occur at an early step of the alpha₁-adrenergic action. TPA (10^?11–10^?6M) was unable to stimulate glycogenolysis. Urea synthesis, which is stimulated by vasopressin and alpha₁-adrenergic agents, was not stimulated by phorbol ester, neither alone nor in combination with the Ca⁺⁺ ionophore A-23187.     

Unusual outcome of in utero infection and subsequent postnatal super-infection with different PCV2b strains

VC2002, isolated from postweaning multisystemic wasting syndrome (PMWS)-affected pig, is a mixture of two porcine circovirus genotype 2b (PCV2b) viruses, K2 and K39. Preliminary experiments disclosed short-term adverse effects of K39, but not K2, on porcine foetuses. These findings led to the hypothesis that infection of immuno-incompetent foetuses with K2 confers a status of immunotolerance, and postnatal super-infection with K39 triggers PMWS. To explore this hypothesis, nine 55-day-old foetuses were inoculated in utero (three with K2-10^4.3TCID₅₀, three with K39-10^4.3TCID₅₀ and three with medium), and foeto-pathogenicity examined. At 21 days post-inoculation (dpi), K2 did not induce pathology, whereas pathological effects of K39 were evident. Twenty-four 45-day-old foetuses were subsequently inoculated to examine the long-term effect of K2, including six with K2-high dose-10^4.3TCID₅₀, six with K2-low dose-10^2.3TCID₅₀ and 12 mock-inoculated controls. Both doses resulted in five mummified foetuses and one live-born piglet each (69dpi). K2 was recovered from all mummies. K2 and K2-specific antibodies were not detected in serum of the two live-born piglets at birth, indicating full control of K2 infection. The K2-low dose-infected piglet was immunostimulated at day 2, but not the K2-high dose-infected piglet. Both non-stimulated and stimulated K2-infected piglets were super-inoculated with K39 at day 6 or 8 (taken as 0 days post super-inoculation). Low viral replication was observed in the non-stimulated K2-K39 piglet (up to 10^3.3TCID₅₀/g; identified as K39). In contrast, viral replication was extremely high in the stimulated K2-K39 piglet (up to 10^5.6TCID₅₀/g) and identified as K2, indicating that K2 infection is controlled during foetal life, but emerges after birth upon immunostimulation. However, none of the piglets showed any signs of PMWS.     

Paenibacillus brassicae sp. nov., isolated from cabbage rhizosphere in Beijing, China

A novel Gram-positive, rod-shaped, motile, spore-forming, nitrogen-fixing bacterium, designated strain 112^T, was isolated from cabbage rhizosphere in Beijing, China. The strain was found to grow at 10–40 °C and pH 4–11, with an optimum of 30 °C and pH 7.0, respectively. Phylogenetic analysis based on a fragment of the full-length 16S rRNA gene sequence revealed that strain 112^T is a member of the genus Paenibacillus. High levels of 16S rRNA gene similarities were found between strain 112^T, Paenibacillus sabinae DSM 17841^T (97.82 %) and Paenibacillus forsythiae DSM 17842^T (97.22 %). However, the DNA–DNA hybridization values between strain 112^T and the type strains of these two species were 10.36 and 6.28 %, respectively. The predominant menaquinone was found to be menaquinone 7 (MK-7). The major fatty acids were determined to be anteiso-C_15:0 and C_16:0. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unknown aminophospholipids. The cell wall peptidoglycan was found to contain meso-diaminopimelic acid. The DNA G+C content was determined to be 55.4 mol%. On the basis of its phenotypic characteristics, 16S rRNA gene sequence analysis and the value of DNA–DNA hybridization, strain 112^T is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus brassicae sp. nov. is proposed. The type strain is 112^T (= ACCC 01125^T = DSM 24983^T).