Self-hypnosis for intrapartum pain management in pregnant nulliparous women: a randomised controlled trial of clinical effectiveness.

OBJECTIVE: (Primary) To establish the effect of antenatal group self-hypnosis for nulliparous women on intra-partum epidural use. DESIGN: Multi-method randomised control trial (RCT). SETTING: Three NHS Trusts. POPULATION: Nulliparous women not planning elective caesarean, without medication for hypertension and without psychological illness. METHODS: Randomisation at 28-32 weeks' gestation to usual care, or to usual care plus brief self-hypnosis training (two × 90-minute groups at around 32 and 35 weeks' gestation; daily audio self-hypnosis CD). Follow up at 2 and 6 weeks postnatal. MAIN OUTCOME MEASURES: Primary: epidural analgesia. Secondary: associated clinical and psychological outcomes; cost analysis. RESULTS: Six hundred and eighty women were randomised. There was no statistically significant difference in epidural use: 27.9% (intervention), 30.3% (control), odds ratio (OR) 0.89 [95% confidence interval (CI): 0.64-1.24], or in 27 of 29 pre-specified secondary clinical and psychological outcomes. Women in the intervention group had lower actual than anticipated levels of fear and anxiety between baseline and 2 weeks post natal (anxiety: mean difference -0.72, 95% CI -1.16 to -0.28, P = 0.001); fear (mean difference -0.62, 95% CI -1.08 to -0.16, P = 0.009) [Correction added on 7 July 2015, after first online publication: 'Mean difference' replaced 'Odds ratio (OR)' in the preceding sentence.]. Postnatal response rates were 67% overall at 2 weeks. The additional cost in the intervention arm per woman was £4.83 (CI -£257.93 to £267.59). CONCLUSIONS: Allocation to two-third-trimester group self-hypnosis training sessions did not significantly reduce intra-partum epidural analgesia use or a range of other clinical and psychological variables. The impact of women's anxiety and fear about childbirth needs further investigation.

Analogous pleiotropic effects of insecticide resistance genotypes in peach-potato aphids and houseflies.

We show that single-point mutations conferring target-site resistance (kdr) to pyrethroids and DDT in aphids and houseflies, and gene amplification conferring metabolic resistance (carboxylesterase) to organophosphates and carbamates in aphids, can have deleterious pleiotropic effects on fitness. Behavioural studies on peach-potato aphids showed that a reduced response to alarm pheromone was associated with both gene amplification and the kdr target-site mutation. In this species, gene amplification was also associated with a decreased propensity to move from senescing leaves to fresh leaves at low temperature. Housefly genotypes possessing the identical kdr mutation were also shown to exhibit behavioural differences in comparison with susceptible insects. In this species, resistant individuals showed no positional preference along a temperature gradient while susceptible genotypes exhibited a strong preference for warmer temperatures.

Purification of matrix Gla protein from a marine teleost fish, Argyrosomus regius: calcified cartilage and not bone as the primary site of MGP accumulation in fish.

Matrix Gla protein (MGP) belongs to the family of vitamin K-dependent, Gla-containing proteins, and in mammals, birds, and Xenopus, its mRNA was previously detected in extracts of bone, cartilage, and soft tissues (mainly heart and kidney), whereas the protein was found to accumulate mainly in bone. However, at that time, it was not evaluated if this accumulation originated from protein synthesized in cartilage or in bone cells because both coexist in skeletal structures of higher vertebrates and Xenopus. Later reports showed that MGP also accumulated in costal calcified cartilage as well as at sites of heart valves and arterial calcification. Interestingly, MGP was also found to accumulate in vertebra of shark, a cartilaginous fish. However, to date, no information is available on sites of MGP expression or accumulation in teleost fishes, the ancestors of terrestrial vertebrates, who have in their skeleton mineralized structures with both bone and calcified cartilage. To analyze MGP structure and function in bony fish, MGP was acid-extracted from the mineralized matrix of either bone tissue (vertebra) or calcified cartilage (branchial arches) from the bony fish, Argyrosomus regius, separated from the mineral phase by dialysis, and purified by Sephacryl S-100 chromatography. No MGP was recovered from bone tissue, whereas a protein peak corresponding to the MGP position in this type of gel filtration was obtained from an extract of branchial arches, rich in calcified cartilage. MGP was identified by N-terminal amino acid sequence analysis, and the resulting protein sequence was used to design specific oligonucleotides suitable to amplify the corresponding DNA by a mixture of reverse transcription-polymerase chain reaction (RT-PCR) and 5'rapid amplification of cDNA (RACE)-PCR. In parallel, ArBGP (bone Gla protein, osteocalcin) was also identified in the same fish, and its complementary DNA cloned by an identical procedure. Tissue distribution/accumulation was analyzed by Northern blot, in situ hybridization, and immunohistochemistry. In mineralized tissues, the MGP gene was predominantly expressed in cartilage from branchial arches, with no expression detected in the different types of bone analyzed, whereas BGP mRNA was located in bone tissue as expected. Accordingly, the MGP protein was found to accumulate, by immunohistochemical analysis, mainly in the extracellular matrix of calcified cartilage. In soft tissues, MGP mRNA was mainly expressed in heart but in situ hybridization, indicated that cells expressing the MGP gene were located in the bulbus arteriosus and aortic wall, rich in smooth muscle and endothelial cells, whereas no expression was detected in the striated muscle myocardial fibers of the ventricle. These results show that in marine teleost fish, as in mammals, the MGP gene is expressed in cartilage, heart, and kidney tissues, but in contrast with results obtained in Xenopus and higher vertebrates, the protein does not accumulate in vertebra of non-osteocytic teleost fish, but only in calcified cartilage. In addition, our results also indicate that the presence of MGP mRNA in heart tissue is due, at least in fish, to the expression of the MGP gene in only two specific cell types, smooth muscle and endothelial cells, whereas no expression was found in the striated muscle fibers of the ventricle. In light of these results and recent information on expression of MGP gene in these same cell types in mammalian aorta, it is likely that the levels of MGP mRNA previously detected in Xenopus, birds, and mammalian heart tissue may be restricted to regions rich in smooth muscle and endothelial cells. Our results also emphasize the need to re-evaluate which cell types are involved in MGP gene expression in other soft tissues and bring further evidence that fish are a valuable model system to study MGP gene expression and regulation.

Characterization of acetylcholinesterases, and their genes, from the hemipteran species Myzus persicae (Sulzer), Aphis gossypii (Glover), Bemisia tabaci (Gennadius) and Trialeurodes vaporariorum (Westwood).

Gene sequences encoding putative acetylcholinesterases have been reported for four hemipteran insect species. Although acetylcholinesterase insensitivity occurs in insecticide-resistant populations of each of these species, no mutations were detected in the gene sequences from the resistant insects. This, coupled with a series of experiments using novel reversible inhibitors to compare the biochemical characteristics of acetylcholinesterase from a range of insect species, showed that the cloned cDNA fragments are unlikely to encode the hemipteran synaptic acetylcholinesterases, and there is likely to be a second ace locus.

Men and osteoporosis.

BACKGROUND: While strict criteria have been developed for defining osteoporosis in women (bone mineral density measurements more than 2.5 standard deviations below the mean for young adult normal women, i.e. t-score value < -2.5), there still remains a controversy regarding the definition in men. Spinal fractures occur in 5% and hip fractures in 6% of men older than 50 years. There are significant differences between men and women with respect to the pathogenesis of osteoporosis, underlying medical conditions and postfracture sequelae. OBJECTIVE: To provide an overview of the pathogenesis, diagnosis and prevention of osteoporosis in men. DISCUSSION: Osteoporosis is increasingly recognised. Data from the Dubbo Osteoporosis Epidemiology Study suggests that 30% of men in Australia aged over 60 years will suffer from an osteoporotic fracture. It is estimated that 30-60% of men presenting with spinal fractures will have another illness contributing to their bone loss. Osteoporotic fractures in men are associated with higher morbidity and mortality than in women. Lifestyle changes together with daily calcium supplementation should be implemented and vitamin D3 should be considered in men with osteopenia.

Matrix Gla protein in Xenopus laevis: molecular cloning, tissue distribution, and evolutionary considerations.

Matrix Gla protein (MGP) belongs to the family of vitamin K-dependent, Gla-containing proteins and in higher vertebrates, is found in the extracellular matrix of mineralized tissues and soft tissues. MGP synthesis is highly regulated at the transcription and posttranscription levels and is now known to be involved in the regulation of extracellular matrix calcification and maintenance of cartilage and soft tissue integrity during growth and development. However, its mode of action at the molecular level remains unknown. Because there is a large degree of conservation between amino acid sequences of shark and human MGP, the function of MGP probably has been conserved throughout evolution. Given the complexity of the mammalian system, the study of MGP in a lower vertebrate might be advantageous to relate the onset of MGP expression with specific events during development. Toward this goal, MGP was purified from Xenopus long bones and its N-terminal amino acid sequence was determined and used to clone the Xenopus MGP complementary DNA (cDNA) by a mixture of reverse-transcription (RT)- and 5'- rapid amplification of cDNA ends (RACE)-polymerase chain reaction (PCR). MGP messenger RNA (mRNA) was present in all tissues analyzed although predominantly expressed in Xenopus bone and heart and its presence was detected early in development at the onset of chondrocranium development and long before the appearance of the first calcified structures and metamorphosis. These results show that in this system, as in mammals, MGP may be required to delay or prevent mineralization of cartilage and soft tissues during the early stages of development and indicate that Xenopus is an adequate model organism to further study MGP function during growth and development.

Identification and characterization of mutations in housefly (Musca domestica) acetylcholinesterase involved in insecticide resistance.

Acetylcholinesterase (AChE) insensitive to organophosphate and carbamate insecticides has been identified as a major resistance mechanism in numerous arthropod species. However, the associated genetic changes have been reported in the AChE genes from only three insect species; their role in conferring insecticide insensitivity has been confirmed, using functional expression, only for those in Drosophila melanogaster. The housefly, Musca domestica, was one of the first insects shown to have this mechanism; here we report the occurrence of five mutations (Val-180-->Leu, Gly-262-->Ala, Gly-262-->Val, Phe-327-->Tyr and Gly-365-->Ala) in the AChE gene of this species that, either singly or in combination, confer different spectra of insecticide resistance. The baculovirus expression of wild-type and mutated housefly AChE proteins has confirmed that the mutations each confer relatively modest levels of insecticide insensitivity except the novel Gly-262-->Val mutation, which results in much stronger resistance (up to 100-fold) to certain compounds. In all cases the effects of mutation combinations are additive. The mutations introduce amino acid substitutions that are larger than the corresponding wild-type residues and are located within the active site of the enzyme, close to the catalytic triad. The likely influence of these substitutions on the accessibility of the different types of inhibitor and the orientation of key catalytic residues are discussed in the light of the three-dimensional structures of the AChE protein from Torpedo californica and D. melanogaster.

Molecular cloning, genomic organization, and expression of a C-type (Manduca sexta-type) allatostatin preprohormone from Drosophila melanogaster.

The insect allatostatins are a diverse group of neuropeptides that obtained their names by their inhibitory actions on the corpora allata (two endocrine glands near the insect brain), where they block the biosynthesis of juvenile hormone (a terpenoid important for development and reproduction). Chemically, the allatostatins can be subdivided into three different peptide groups: the large group of A-type (cockroach-type) allatostatins, which have the common C-terminal sequence Y/FXFGLamide; the B-type (cricket-type) allatostatins, which have the C-terminal sequence W(X(6))Wamide in common; and a single allatostatin that we now call C-type allatostatin that was first discovered in the moth Manduca sexta, and which has a nonamidated C terminus, and a structure unrelated to the A- and B-type allatostatins. We have previously cloned the preprohormones for the A- and B-type allatostatins from Drosophila melanogaster. Here we report on the cloning of a Drosophila C-type allatostatin preprohormone (DAP-C). DAP-C is 121 amino acid residues long and contains one copy of a peptide sequence that in its processed form has the sequence Y in position 4) from the Manduca sexta C-type allatostatin. The DAP-C gene has three introns and four exons and is located at position 32D2-3 on the left arm of the second chromosome. Northern blots show that the gene is strongly expressed in larvae and adult flies, but less in pupae and embryos. In situ hybridizations of larvae show that the gene is expressed in various neurons of the brain and abdominal ganglia and in endocrine cells of the midgut. This is the first publication on the structure of a C-type allatostatin from insects other than moths and the first report on the presence of all three types of allatostatins in a representative of the insect order Diptera (flies).

New graduate RN internship program: a collaborative approach for system-wide integration.

Population growth and aging increase the demand for skilled nurses. Hospitals face the challenge of supplying experienced and qualified caregivers. The cost of training new nurses remains significant. Nursing educators are pivotal in developing the nursing workforce, especially under tightening constraints. This article examines the development of a nursing internship program aimed at attracting and retaining newly graduated nurses and its effect on retention to the institution.

Dealing with diversity: incorporating cultural sensitivity into professional midwifery practice.

In the Australian College of Midwives, Code of Ethics, Section 11. Practice of Midwifery, the following is stated "A. Midwives provide care for women and childbearing families with respect for cultural diversity while also working to eliminate harmful practices within those same cultures." However, it is difficult to know what is meant by "respect for cultural diversity". This paper presents the results of a critical review of the health literature. There is surprisingly little consensus about the meaning of terms such as cultural sensitivity and cultural appropriate care. Nor are there reflections on incorporating these concepts into practice. It could be argued that until there is greater clarity about these concepts and more discussion of how they may be used in practice, midwives would have to continue to rely on their individual knowledge and experience.

Cellular and humoral factors in colostrum of HIV infected and uninfected lactating mothers.

OBJECTIVE: To compare the cellular and humoral factors in colostrum from HIV infected and uninfected lactating mothers. DESIGN: Cross sectional study. SETTING: Maternity Ward. METHODS: Colostrum was collected from 130 mothers (62 HIV seropositives and 68 HIV seronegatives). These colostrum samples were tested for total cell count, cell viability, differential count, phagocytic activity of macrophages, 'T' cell counts, IgA, IgM and IgG levels. RESULTS: There was a statistically significant decrease in the phagocytosis and 'T' cell number (p <0.001) and in the IgA and IgG levels (p<0. 05) in the colostrum obtained from HIV seropositive mothers as compared to HIV seronegative ones. CONCLUSION: Some of the cellular and humoral factors are reduced in colostrum samples obtained from HIV seropositives as compared to normals.

Molecular characterization of a neuronal low-voltage-activated T-type calcium channel.

The molecular diversity of voltage-activated calcium channels was established by studies showing that channels could be distinguished by their voltage-dependence, deactivation and single-channel conductance. Low-voltage-activated channels are called 'T' type because their currents are both transient (owing to fast inactivation) and tiny (owing to small conductance). T-type channels are thought to be involved in pacemaker activity, low-threshold calcium spikes, neuronal oscillations and resonance, and rebound burst firing. Here we report the identification of a neuronal T-type channel. Our cloning strategy began with an analysis of Genbank sequences defined as sharing homology with calcium channels. We sequenced an expressed sequence tag (EST), then used it to clone a full-length complementary DNA from rat brain. Northern blot analysis indicated that this gene is expressed predominantly in brain, in particular the amygdala, cerebellum and thalamus. We mapped the human gene to chromosome 17q22, and the mouse gene to chromosome 11. Functional expression of the channel was measured in Xenopus oocytes. Based on the channel's distinctive voltage dependence, slow deactivation kinetics, and 7.5-pS single-channel conductance, we conclude that this channel is a low-voltage-activated T-type calcium channel.

Effect of storage of colostrum in various containers.

OBJECTIVE: To compare the effect of storage on expressed colostrum kept at room and refrigeration temperature in 2 different types of containers (steel and plastic) at different time intervals (0 hours to 7 hours). DESIGN: Prospective immunological study. SETTING: Maternity ward. METHODS: Colostrum was collected from 60 healthy lactating mothers and tested for total and differential cell counts and cell viability in plastic (polypropylene) and steel containers at 0 hours and 7 hours after storage at 28 degrees C and 4 degrees C. RESULTS: Colostrum stored for 7 hours in plastic containers had a significantly (p < 0.001) higher cell count and viability compared to that stored in steel containers both at 4 degrees C and 28 degrees C. The differential cell count did not vary with time, temperature or storage vessel. CONCLUSION: If required, colostrum should be stored at 4 degrees C in plastic (polypropylene) containers to maintain its protective quality.

Characterization of the acetylcholinesterase gene from insecticide-resistant houseflies (Musca domestica).

Acetylcholinesterase (AChE) is the target site for the organophosphates and carbamates in insects. Widespread use of these two classes of insecticides has led to the selection of resistance. Target modification was regarded as a molecular mechanism in some resistance species. The altered AChEs with reduced sensitivity to inhibition are related to this resistance. AChE genes from two insecticide-resistant housefly (Musca domestica) strains D3 and Kash were isolated and sequenced using RT-PCR and streptavidin-linked magnetic bead techniques. The cDNAs have a 2082-bp open reading frame from which the complete amino acid sequence of AChE has been deduced. Some differences in nucleotide sequence and four-point mutations of amino acid were found compared to a susceptible strain, i.e., the Cooper strain. Three substitutions are likely to confer insecticide insensitivity, which seems that D3 and Kash belong to CH2 pattern of resistance.

Effects of lifelong consumption of fluoridated water or use of fluoride supplements on dental caries prevalence.

This study was undertaken to determine the prevalence of dental caries in children ages 6-14, from fluoridated and non-fluoridated areas in British Columbia, Canada, and the effects of receiving certain fluoride preventive procedures during childhood. Children from two communities were surveyed using a modified Decayed, Missing and Filled Tooth Surface Index (D1D2MFS). Questionnaires on the use of various fluoride preventive practices and residence histories during childhood were collected. Completed questionnaires were returned and exams were performed on 1131 children. Crude caries prevalence scores for the different fluoride exposure groups were tested for differences in dental age and the level of educational attainment of parents and/or guardians. No significant group differences were found. The 110 children with lifelong exposure only to fluoridated water had 35%, or 0.88 (S.D. = 2.91), fewer decayed or filled tooth surfaces per child (P < 0.07) than children with no reported exposure to systemic fluorides. For the 122 children who had taken fluoride supplements for 4 yr or more, 0.67 fewer decayed and filled tooth surfaces (26% reduction) were observed per child when compared to children with no exposure to fluoridated water or supplements. For children who used fluoride supplements for less than 4 yr, no significant benefits were observed. Approximately 75% of the caries prevalence for control and fluoride-exposed groups was on pit and fissured surfaces. Reductions by surface type showed savings on both smooth and pit and fissured surfaces.

Polyphenols, astringency and proline-rich proteins.

Recent, NMR and precipitation, studies of molecular recognition of proline-rich proteins and peptides by plant polyphenols are described and rationalized. The action of polysaccharides and caseins in the moderation of the astringent response, which is engendered by polyphenols present in foodstuffs and beverages, is described. The possible influence of plant cell wall glycoproteins on the process of lignification is discussed in the light of the observed affinity of phenolic substrates for prolyl residues in protein structures.

Functional analysis of the promoter region of a nodule-enhanced glutamine synthetase gene from Phaseolus vulgaris L.

The 5'-flanking region of gln-gamma, the nodule-enhanced glutamine synthetase gene from Phaseolus vulgaris L., has been analysed for cis-regulatory elements using a series of 5' deletions and hybrid gln-gamma:: CaMV 35S promoters. The promoters were fused to the uidA reporter gene and their activities tested in two heterologous expression systems. In the first system, the chimaeric genes were transferred to Lotus corniculatus L. using Agrobacterium rhizogenes and their expression was studied in nodulated hairy roots. In the second system, the constructs were electroporated into tobacco mesophyll protoplasts. The results of the 5' deletion analysis showed that the sequence between -597 and -21 (relative to the ATG codon) was sufficient for nodule-specific expression of the chimaeric gene in nodulated hairy roots, and revealed the existence of at least two positive regulatory elements. Sequences located between -2000 and -597 were able to stimulate expression in nodules but not protoplasts, while the region from -597 to -354 enhanced expression in both nodules and protoplasts. Results obtained with the hybrid gln-gamma::35S promoters showed that two overlapping restriction fragments (-516/-343 and -474/-293) were able to stimulate expression from a heterologous promoter in an orientation-dependent manner. Previous work has demonstrated the presence of conserved A/T-rich binding sites for nuclear proteins in the region between -516 and -446, and their possible role in regulating gln-gamma expression is discussed.