The immediate effects of cervical spine manipulation on pain and biochemical markers in females with acute non-specific mechanical neck pain: a randomized clinical trial.

Study Design: Randomized clinical trial with pre-test, post-test control group design. Objectives: To examine the immediate effects of cervical spinal manipulation (CSM) on serum concentration of biochemical markers (oxytocin, neurotensin, orexin A, and cortisol). Background: Several studies have found an association between spinal manipulation (SM) and pain perception. However, the mechanism by which SM modulates pain remains undefined. Methods: Twenty-eight female subjects with non-specific mechanical neck pain were randomly assigned to one of two interventions (CSM versus sham CSM). Blood samples were drawn before and immediately after the respective interventions. Oxytocin, neurotensin, orexin A, and cortisol were measured from the blood and serum using the Milliplex Map Magnetic Bead Panel Immunoassay on the Luminex 200 Platform. Results: In the CSM group, there were significant increases in pre- versus post-manipulation mean oxytocin (154.5 ± 60.1 vs. 185.1 ± 75.6, p = .012); neurotensin (116.0 ± 26.5 vs.136.4 ± 34.1, p < . 001); orexin A (52.2 ± 31.1 vs. 73.8 ± 38.8, p < .01) serum concentration; but no significant differences in mean cortisol (p = .052) serum concentration. In the sham group, there were no significant differences in any of the biomarkers (p > .05). Conclusion: The results of the current study suggest that the mechanical stimuli provided through a CSM may modify neuropeptide expression by immediately increasing the serum concentration of nociception-related biomarkers (oxytocin, neurotensin, orexin A, but not cortisol) in the blood of female subjects with non-specific mechanical neck pain.

Study of seleniferous soils using instrumental neutron activation analysis.

Soil samples from the seleniferous region of Punjab State in India were analyzed by instrumental neutron activation analysis (INAA) using reactor neutrons and high resolution γ-ray spectrometry. Samples were collected from three different depths namely surface, root and geological bed zones. Concentrations of 15 elements including selenium and arsenic were determined by relative method. For comparison purposes, soil samples collected from a non-seleniferous region were also analyzed.

Gene doping detection: evaluation of approach for direct detection of gene transfer using erythropoietin as a model system.

As clinical gene therapy has progressed toward realizing its potential, concern over misuse of the technology to enhance performance in athletes is growing. Although 'gene doping' is banned by the World Anti-Doping Agency, its detection remains a major challenge. In this study, we developed a methodology for direct detection of the transferred genetic material and evaluated its feasibility for gene doping detection in blood samples from athletes. Using erythropoietin (EPO) as a model gene and a simple in vitro system, we developed real-time PCR assays that target sequences within the transgene complementary DNA corresponding to exon/exon junctions. As these junctions are absent in the endogenous gene due to their interruption by introns, the approach allows detection of trace amounts of a transgene in a large background of the endogenous gene. Two developed assays and one commercial gene expression assay for EPO were validated. On the basis of ability of these assays to selectively amplify transgenic DNA and analysis of literature on testing of gene transfer in preclinical and clinical gene therapy, it is concluded that the developed approach would potentially be suitable to detect gene doping through gene transfer by analysis of small volumes of blood using regular out-of-competition testing.

Dry heat, moist heat and body fat: are heating modalities really effective in people who are overweight?

Surface heating modalities are commonly used in physical therapy and physical medicine for increasing circulation, especially in deep tissues, to promote healing. However, recent evidence seems to indicate that in people who are overweight, heat transfer is impaired by the subcutaneous fat layer. The present investigation was conducted on 10 subjects aged 22-54 years, whose body mass index averaged 25.8+/-4.6. Subcutaneous fat above the quadriceps muscle varied from 0.51 to 0.86 cm of thickness. Three heating modalities were examined: the application of dry heat with a commercial chemical heat pack, hydrocollator heat packs (providing a type of moist heat), and a whirlpool, where conductive heat loss through water contact would be very high. The temperature of the skin and the temperature in the muscle (25 mm below the skin surface) were assessed by thermocouples. The results of the experiments showed that for heating modalities that are maintained in skin contact for long periods of time, such as dry heat packs (in place for 6 hours), subcutaneous fat did not impair the change in deep muscle temperature. In contrast, when rapid heat modalities were used, such as the hydrocollator and the whirlpool (15 minutes of sustained skin contact), the transfer of heat from the skin to deep muscle was significantly impaired in people with thicker subcutaneous fat layers. We observed that the greater the impairment in heat transfer to muscle from skin covered by body fat, the warmer the skin temperature increase during the modality.

Does skin moisture influence the blood flow response to local heat? A re-evaluation of the Pennes model.

Pennes first described a model of heat transfer through the limb based only on calories delivered from a heat source, calories produced by metabolism and skin blood flow. The purpose of this study was to determine the effect of a moist versus a dry heat source on the skin in eliciting a blood flow response to add data to this model. Ten subjects were examined, both male and female, with a mean age of 32.5 +/- 11.6 years, mean height of 172.8 +/- 12.3 cm, and mean weight of 77.6 +/- 19.5 kg. Skin temperature was measured by a thermocouple placed on the skin and skin blood flow measured by a laser Doppler flow meter. The results of the experiments using a dry heat pack (commercially available chemical 42 degrees C cell dry heat source), moist hydrocollator pack (72.8 degrees C) separated from the skin by eight layers of towels, and whirlpool at 40 degrees C, showed that moist heat caused a significantly higher skin blood flow (about 500% greater) than dry heat (p < 0.01). Most of the greater increase in skin blood flow with moist heat was due to the greater rate of rise of skin temperature with moist versus dry heat while some of the increase in blood flow was due to the moisture itself. This could either be related to the greater heat flux across the skin with moist air or due to changing the ionic environment around skin thermo receptors by keeping the skin moist during heating. Skin thermo receptors are believed to be temperature sensitive calcium gated channels in endothelial cells which couple calcium influx to a release of nitric oxide. If true, reducing moisture in the skin might have the effect of altering ionic flux through these receptors. A correct model of skin heat flux should therefore take heat moisture content into consideration.

Crystal structure of the hexameric replicative helicase RepA of plasmid RSF1010.

Unwinding of double-stranded DNA into single-stranded intermediates required for various fundamental life processes is catalyzed by helicases, a family of mono-, di- or hexameric motor proteins fueled by nucleoside triphosphate hydrolysis. The three-dimensional crystal structure of the hexameric helicase RepA encoded by plasmid RSF1010 has been determined by X-ray diffraction at 2.4 A resolution. The hexamer shows an annular structure with 6-fold rotational symmetry and a approximately 17 A wide central hole, suggesting that single-stranded DNA may be threaded during unwinding. Homologs of all five conserved sequence motifs of the DnaB-like helicase family are found in RepA, and the topography of the monomer resembles RecA and the helicase domain of the bacteriophage T7 gp4 protein. In a modeled complex, ATP molecules are located at the subunit interfaces and clearly define adenine-binding and ATPase catalytic sites formed by amino acid residues located on adjacent monomers; most remarkable is the "arginine finger" Arg207 contributing to the active site in the adjacent monomer. This arrangement of active-site residues suggests cooperativity between monomers in ATP hydrolysis and helicase activity of RepA. The mechanism of DNA unwinding remains elusive, as RepA is 6-fold symmetric, contrasting the recently published asymmetric structure of the bacteriophage T7 gp4 helicase domain.

The Tom channel in the mitochondrial outer membrane: alive and kicking.

The targeting of newly-made polypeptides to specific membranes, and the subsequent ability of a membrane to allow only certain polypeptides into its compartment, are essential to maintain the ultrastructure of Eukaryotic cells. Distinct oligomeric protein complexes in each cellular membrane catalyse these translocation processes. A recent report [Hill K et al. Nature 1998;395:516-521 (Ref. 1)] of the reconstitution of the translocation channel from the mitochondrial outer membrane, after producing the major structural component of the channel by recombinant means, promises a system to dissect in molecular detail the exact working of one of these protein translocation machines.

The three-dimensional structure of bovine odorant binding protein and its mechanism of odor recognition.

Odorant binding protein (OBP) is the major odorant binding component of mammalian nasal mucosa. The two structures of bovine OBP reported in this paper (one crystallized as purified and one soaked in the presence of a selenium-containing odorant) show that: (i) the OBP dimer is composed of two compact domains related by an approximate two-fold axis of symmetry; (ii) between residues 122 and 123 the polypeptide chains cross from one domain to the other such that each domain is formed by residues from both monomers; (iii) purified OBP already contains two bound odorant molecules (one per monomer)-odorant binding occurs by replacement of these molecules with the added odorant; and (iv) the structure of the odorant binding site can explain OBP's extraordinarily broad odorant specificity.

Microcalorimetric study of wheat germ agglutinin binding to N-acetylglucosamine and its oligomers.

The energetics of association of wheat germ agglutinin (WGA) with N-acetylglucosamine (GlcNAc) and its beta(1,4) oligomers have been measured using isothermal titration calorimetry. Association constants of 0.4, 5.3, 11.1, 12.3, and 19.1 mM-1 and enthalpies of binding of -6.1, -15.6, -19.4, -19.3, and -18.2 kcal mol-1 were obtained at 26 degrees C for the titration of WGA with GlcNAc, (GlcNAc)2, (GlcNAc)3, (GlcNAc)4, and (GlcNAc)5, respectively. The term T delta S was always of negative value, indicating that the binding process is enthalpically driven. Titrations of WGA performed at pH 4.5 did not differ significantly from those performed at pH 7.0, suggesting that no groups with a pKa in this range are directly involved in the binding event. Also, performing the titration in a buffer system with a higher enthalpy of protonation did not change the enthalpy of binding confirming that there is no net protonation or deprotonation when WGA binds GlcNAc residues at pH 7. A model of four independent binding sites was found to adequately describe the binding curves, except in the case of (GlcNAc)4 which exhibited positive cooperativity. The energetic values are discussed within the context of the structure of the WGA-(GlcNAc)2 complex.

Late effects of cyclophosphamide treatment of neonatal mice.

Swiss mice surviving early onset of wasting disease at 4-6 weeks following cyclophosphamide administration at birth suffer from delayed effects of this immunosuppressive drug. The late wasting syndrome developing at 6-8 months post-inoculation is characterized clinically by loss of weight, hunched posture, ruffled fur and diarrhoea. Lymphocyte and granulocyte levels are raised. The lymphocyte/granulocyte ratio is significantly inhibited. The development of various pathological lesions in thymus, spleen, lymph nodes and bone-marrow is frequently observed. Infiltration of lymphoid tissue in lungs, liver and kidneys is a common feature. It is hoped that further experimental studies would provide more insight into the delayed adverse effects of cyclophosphamide therapy.

Pathogenesis of wasting disease after cyclophosphamide treatment of neonatal mice.

Cyclophosphamide is a potent immunosuppressive agent and is being widely used in organ transplantation. The effects of this anti-rejection drug on lymphoid organs are poorly understood. Newborn Swiss mice injected with various doses of cyclophosphamide suffered from wasting disease at 4 weeks post treatment. The incidence of wasting disease was dose dependent. Haematological picture of the wasting animals revealed leukocytosis of variable degree. Lymphocyte/granulocyte ratio was not inhibited. The cyclophosphamide treatment caused shrinkage of lymphoid organs. Bone marrow showed degeneration of haematopoietic cells. The failure to sustain lymphopoiesis by the potential lymphoid sites following cyclophosphamide treatment and the associated immunological insufficiency resulted in the fatal wasting disease.

Calorimetric determination of cooperative interactions in high affinity binding processes.

It is demonstrated that isothermal titration calorimetry can be used to determine cooperative interaction energetics even for extremely tight binding processes in which the binding affinity constants are beyond the limits of experimental determination. The approach is based on the capability of calorimetry to measure the apparent binding enthalpy at any degree of ligand saturation. When calorimetric measurements are performed under conditions of total association at partial saturation, the dependence of the apparent binding enthalpy on the degree of saturation is a function only of the cooperative binding interactions. The method developed in this paper allows an independent estimation of cooperative energetic parameters without the need to simultaneously estimate or precisely know the value of the association constants. Since total ligand association at partial saturation is achieved only at macromolecular concentrations much larger than the dissociation constants, the method is especially suited for high and very high affinity processes. Biological associations in this category include fundamental cellular processes like cell surface receptor binding or protein-DNA interactions.

Attributional patterns, moods, and the menstrual cycle.

Sixty women attending a family planning clinic completed assessments of their attributions for negative and positive mood states for an individual at different menstrual cycle stages. Negative moods occurring premenstrually were almost invariably viewed as related to health factors. Positive moods tended to be attributed to environmental events and the person's lifestyle details. Personality tended to be used as an attribution for moods occurring intermenstrually. There was no association between attributional patterns and menstrual cycle complaints or attitudes to the menstrual cycle as measured by the Menstrual Attitude Questionnaire. The results are discussed within the framework of the social cognition theory of menstrual cycle complaints.

Effect of extrusion processing on the nutritional quality of protein in rice-legume blends.

The effect of extrusion processing using Wenger X-5 extruder on the quality of products of rice-legume (75:25) blends was investigated. The products were palatable and had a good texture. The nutritional quality of protein in the extrudates was significantly improved. The protein efficiency ratio values of rice-soybean, rice-bengal gram and rice-black gram products were: 2.25, 2.30 and 2.28 as compared to 2.10, 1.89 and 1.98 for the respective raw blends. Extrusion processing reduced the phytates in the products to the extent of 20.3 to 26.8%.

Effects of neonatal thymic exposure to high doses of X-irradiation.

The thymic region of neonatal Swiss mice was exposed to doses varying from 1000 R to 2000 R of X-irradiation. The animals did not show any signs of wasting syndrome up to 6 months after irradiation. At this time hyperplasia of the thymus with an associated lymphocytosis was evident in irradiated animals. Antibody production to sheep red blood cells (SRBC) was not affected. However, at 12 months post-irradiation the animals showed signs of wasting disease with a progressive increase in their numbers at 18 and 24 months of age. The percentage incidence of animals with wasting disease was dose dependent. At this stage in the majority of the animals with the disease the thymus showed varying degrees of atrophy along with splenomegaly. There were no significant differences in the number of lymphocytes but the number of granulocytes showed a substantial increase. This was more evident in animals exposed to 2000 R to the thymic region. Though one observed a lowered ability to form antibodies to bovine serum albumin (BSA) with advancing age, the thymic irradiation did not affect the immune response to BSA even in animals manifesting wasting disease. An interesting observation has been the development of a severe loss of muscle power and tone in the hind limbs in a large majority of animals.