Functional role of histamine receptors in the renal cortical collecting duct cells.
Histamine is an important immunomodulator, as well as a regulator of allergic inflammation, gastric acid secretion, and neurotransmission. Although substantial histamine level has been reported in the kidney, renal pathological and physiological effects of this compound have not been clearly defined. The goal of this study was to provide insight into the role of histamine-related pathways in the kidney, with emphasis on the collecting duct (CD), a distal part of the nephron important for the regulation of blood pressure. We report that all four histamine receptors (HRs) as well as enzymes responsible for histamine metabolism and synthesis are expressed in cultured mouse mpkCCDcl4 cells, and histamine evokes a dose-dependent transient increase in intracellular Ca2+ in these cells. Furthermore, we observed a dose-dependent increase in cAMP in the CD cells in response to histamine. Short-circuit current studies aimed at measuring Na+ reabsorption via ENaC (epithelial Na+ channel) demonstrated inhibition of ENaC-mediated currents by histamine after a 4-h incubation, and single-channel patch-clamp analysis revealed similar ENaC open probability before and after acute histamine application. The long-term (4 h) effect on ENaC was corroborated in immunocytochemistry and qPCR, which showed a decrease in protein and gene expression for αENaC upon histamine treatment. In summary, our data highlight the functional importance of HRs in the CD cells and suggest potential implications of histamine in inflammation-related renal conditions. Further research is required to discern the molecular pathways downstream of HRs and assess the role of specific receptors in renal pathophysiology.


Identification of a lead small-molecule inhibitor of the Aurora kinases using a structure-assisted, fragment-based approach.
Aurora A and Aurora B are potential targets for anticancer drug development due to their roles in tumorigenesis and disease progression. To identify small-molecule inhibitors of the Aurora kinases, we undertook a structure-based design approach that used three-dimensional structural models of the Aurora A kinase and molecular docking simulations of chemical entities. Based on these computational methods, a new generation of inhibitors derived from quinazoline and pyrimidine-based tricyclic scaffolds were synthesized and evaluated for Aurora A kinase inhibitory activity, which led to the identification of 4-(6,7-dimethoxy-9H-1,3,9-triaza-fluoren-4-yl)-piperazine-1-carbothioic acid [4-(pyrimidin-2-ylsulfamoyl)-phenyl]-amide. The lead compound showed selectivity for the Aurora kinases when it was evaluated against a panel of diverse kinases. Additionally, the compound was evaluated in cell-based assays, showing a dose-dependent decrease in phospho-histone H3 levels and an arrest of the cell cycle in the G(2)-M fraction. Although biological effects were observed only at relatively high concentrations, this chemical series provides an excellent starting point for drug optimization and further development.


[Effect of rations containing fluorine ion on biochemical and hematological indices of body status].
A study was made of the fluorine-ion content in the vegetables and grain grown on experimental plots of land fertilized for 4-6 years with fluorine-containing fertilizers: active slurry potassium tetrafluoroborate and sodium fluorosilicate. It was found that the fluorine-ion content in individual samples of the vegetables and grain was increased 2-32-fold as compared to that in the control samples. As for the diet, the total content of fluorine-ion was increased 6.1-12.3-fold. Experiments were made on rabbits and rats whose diets contained the vegetables and grain from the experimental plots of land. The animals of the experimental groups manifested a lowering of the prothrombin index by the end of the 6th month of the experimental studies. There was a statistically significant fall in the cholinesterase activity and an increase in the activity of glutaminopyruvic aminotransferase (by 35-42 units). The activity of total alkaline phosphatase progressively increased at the expense of the elevated activity of liver and intestinal isozymes.


Cloning and expression of a cDNA encoding human sterol carrier protein 2.
We report the cloning and expression of a cDNA encoding human sterol carrier protein 2 (SCP2). The 1.3-kilobase (kb) cDNA contains an open reading frame which encompasses a 143-amino acid sequence which is 89% identical to the rat SCP2 amino acid sequence. The deduced amino acid sequence of the polypeptide reveals a 20-residue amino-terminal leader sequence in front of the mature polypeptide, which contains a carboxyl-terminal tripeptide (Ala-Lys-Leu) related to the peroxisome targeting sequence. The expressed cDNA in COS-7 cells yields a 15.3-kDa polypeptide and increased amounts of a 13.2-kDa polypeptide, both reacting with a specific rabbit antiserum to rat liver SCP2. The cDNA insert hybridizes with 3.2- and 1.8-kb mRNA species in human liver poly(A)+ RNA. In human fibroblasts and placenta the 1.8-kb mRNA was most abundant. Southern blot analysis suggests either that there are multiple copies of the SCP2 gene in the human genome or that the SCP2 gene is very large. Coexpression of the SCP2 cDNA with expression vectors for cholesterol side-chain cleavage enzyme and adrenodoxin resulted in a 2.5-fold enhancement of progestin synthesis over that obtained with expression of the steroidogenic enzyme system alone. These findings are concordant with the notion that SCP2 plays a role in regulating steroidogenesis, among other possible functions.


Temperature and growth phase influence the outer-membrane proteome and the expression of a type VI secretion system in Yersinia pestis.
Yersinia pestis cells were grown in vitro at 26 and 37 degrees C, the ambient temperatures of its flea vector and its mammalian hosts, respectively, and subjected to subcellular fractionation. Abundance changes at 26 vs 37 degrees C were observed for many outer-membrane (OM) proteins. The cell adhesion protein Ail (y1324) and three putative small beta-barrel OM proteins (y1795, y2167 and y4083) were strongly increased at 37 degrees C. The Ail/Lom family protein y1682 (OmpX) was strongly increased at 26 degrees C. Several porins and TonB-dependent receptors, which control small molecule transport through the OM, were also altered in abundance in a temperature-dependent manner. These marked differences in the composition of the OM proteome are probably important for the adaptation of Y. pestis to its in vivo life stages. Thirteen proteins that appear to be part of an intact type VI secretion system (T6SS) were identified in membrane fractions of stationary-phase cells grown at 26 degrees C, but not at 37 degrees C. The corresponding genes are clustered in the Y. pestis KIM gene locus y3658-y3677. The proteins y3674 and y3675 were particularly abundant and co-fractionated in a Mr range indicative of participation in a multi-subunit complex. The soluble haemolysin-coregulated protein y3673 was even more abundant. Its release into the extracellular medium was triggered by treatment of Y. pestis cells with trypsin. Proteases and other stress-response-inducing factors may constitute environmental cues resulting in the activation of the T6SS in Y. pestis.


Snooker: a structure-based pharmacophore generation tool applied to class A GPCRs.
G-protein coupled receptors (GPCRs) are important drug targets for various diseases and of major interest to pharmaceutical companies. The function of individual members of this protein family can be modulated by the binding of small molecules at the extracellular side of the structurally conserved transmembrane (TM) domain. Here, we present Snooker, a structure-based approach to generate pharmacophore hypotheses for compounds binding to this extracellular side of the TM domain. Snooker does not require knowledge of ligands, is therefore suitable for apo-proteins, and can be applied to all receptors of the GPCR protein family. The method comprises the construction of a homology model of the TM domains and prioritization of residues on the probability of being ligand binding. Subsequently, protein properties are converted to ligand space, and pharmacophore features are generated at positions where protein ligand interactions are likely. Using this semiautomated knowledge-driven bioinformatics approach we have created pharmacophore hypotheses for 15 different GPCRs from several different subfamilies. For the beta-2-adrenergic receptor we show that ligand poses predicted by Snooker pharmacophore hypotheses reproduce literature supported binding modes for ∼75% of compounds fulfilling pharmacophore constraints. All 15 pharmacophore hypotheses represent interactions with essential residues for ligand binding as observed in mutagenesis experiments and compound selections based on these hypotheses are shown to be target specific. For 8 out of 15 targets enrichment factors above 10-fold are observed in the top 0.5% ranked compounds in a virtual screen. Additionally, prospectively predicted ligand binding poses in the human dopamine D3 receptor based on Snooker pharmacophores were ranked among the best models in the community wide GPCR dock 2010.


Oxidation by DNA charge transport damages conserved sequence block II, a regulatory element in mitochondrial DNA.
Sites of oxidative damage in mitochondrial DNA have been identified on the basis of DNA-mediated charge transport. Our goal is to understand which sites in mitochondrial DNA are prone to oxidation at long range and whether such oxidative damage correlates with cancerous transformation. Here we show that a primer extension reaction can be used to monitor directly oxidative damage to authentic mitochondrial DNA through photoreactions with a rhodium intercalator. The complex [Rh(phi)2bpy]Cl3 (phi = 9,10-phenanthrenequinone diimine) binds to DNA without sequence specificity and, upon photoactivation, either promotes strand breaks directly at the binding site or promotes one-electron oxidative damage; comparing the sites of base oxidation to direct strand breaks reveals the oxidative damage that arises from a distance through DNA-mediated charge transport. Significantly, base oxidation by charge transport overlaps with known mutational hot spots associated with cancers at nucleotides surrounding positions 263 and 303; the latter is known as conserved sequence block II and is vital to DNA replication. Since DNA base oxidation at conserved sequence block II should weaken the ability of damaged mitochondrial genomes to be replicated, DNA-mediated charge transport may provide a protection mechanism for excluding damaged DNA.


A basal alvarezsauroid theropod from the early Late Jurassic of Xinjiang, China.
The fossil record of Jurassic theropod dinosaurs closely related to birds remains poor. A new theropod from the earliest Late Jurassic of western China represents the earliest diverging member of the enigmatic theropod group Alvarezsauroidea and confirms that this group is a basal member of Maniraptora, the clade containing birds and their closest theropod relatives. It extends the fossil record of Alvarezsauroidea by 63 million years and provides evidence for maniraptorans earlier in the fossil record than Archaeopteryx. The new taxon confirms extreme morphological convergence between birds and derived alvarezsauroids and illuminates incipient stages of the highly modified alvarezsaurid forelimb.


Who does not participate in a follow-up postal study? a survey of infertile couples treated by in vitro fertilization.
A good response rate has been considered as a proof of a study's quality. Decreasing participation and its potential impact on the internal validity of the study are of growing interest. Our objective was to assess factors associated with contact and response to a postal survey in a epidemiological study of the long-term outcome of IVF couples. The DAIFI study is a retrospective cohort including 6,507 couples who began an IVF program in 2000-2002 in one of the eight participating French IVF centers. Medical data on all 6,507 couples were obtained from IVF center databases, and information on long-term outcome was available only for participants in the postal survey (n = 2,321). Logistic regressions were used to assess firstly factors associated with contact and secondly factors associated with response to the postal questionnaire among contacted couples. Sixty-two percent of the 6,507 couples were contacted and 58% of these responded to the postal questionnaire. Contacted couples were more likely to have had a child during IVF treatment than non-contactable couples, and the same was true of respondents compared with non-respondents. Demographic and medical characteristics were both associated with probability of contact and probability of response. After adjustment, having a live birth during IVF treatment remained associated with both probabilities, and more strongly with probability of response. Having a child during IVF treatment was a major factor impacting on participation rate. Non-response as well as non-contact were linked to the outcome of interest, i.e. long-term parenthood success of infertile couples. Our study illustrates that an a priori hypothesis may be too simplistic and may underestimate potential bias. In the context of growing use of analytical methods that take attrition into account (such as multiple imputation), we need to better understand the mechanisms that underlie attrition in order to choose the most appropriate method.


On the importance of beta carotene supplementation for calves.
It is evident from the results, that the beta carotene and vitamin A levels in blood serum of calves of different age could be significantly increased by Rovimix--beta carotene supplementation. High utilization rate of beta carotene from Rovimix as well as considerable beta carotene conversion into vitamin A was also found in calves from dams supplemented with Rovimix--beta carotene in the period of late pregnancy. The contribution of beta carotene reserves of the mother to the vitamin A stores of the foetus and to the beta carotene and vitamin A concentrations in blood serum of calves suggest that the main preventive method of beta carotene and vitamin A deficiency in calves should be high intake of beta carotene from daily ration fed to cows or Rovimix--beta carotene supplementation during late pregnancy of cows.


Acylation of lysolecithin in the intestinal mucosa of rats.
1. The presence of an active acyl-CoA-lysolecithin (1-acylglycerophosphorylcholine) acyltransferase was demonstrated in rat intestinal mucosa. 2. ATP and CoA were necessary for the incorporation of free [1-(14)C]oleic acid into lecithin (phosphatidylcholine). 3. The reaction was about 20 times as fast with [1-(14)C]oleoyl-CoA as with free oleic acid, CoA and ATP. 4. With 1-acylglycerophosphorylcholine as the acceptor, both oleic acid and palmitic acid were incorporated into the beta-position of lecithin; the incorporation of palmitic acid was 60% of that of oleic acid. 5. Of the various analogues of lysolecithin tested as acyl acceptors from [1-(14)C]oleoyl CoA, a lysolecithin with a long-chain fatty acid at the 1-position was most efficient. 6. The enzyme was mostly present in the brush-border-free particulate fraction of the intestinal mucosa. 7. Of the various tissues of rats tested for the activity, intestinal mucosa was found to be the most active, with testes, liver, kidneys and spleen following it in decreasing order.


Whistle Abrasion: A Case Report.
Abrasion is the physical wearing of a tooth surface which can involve the presence of a foreign object repeatedly being in contact with the tooth. A 40-year-old male patient reported to our dental clinic with a 2-3 mm uneven gap between his upper and lower front teeth on occlusion. A detailed history revealed that he was a physical education teacher, and the habitual placement of the whistle for the last 15 years caused an indentation on the whistle which coincided with the abraded teeth. Conditions such as abrasion may need active restorations. A general dental practitioner should accurately identify the cause and treat the esthetic and functional impairment as required.


Circumnutation behavior of an exotic honeysuckle vine and its native congener: influence on clonal mobility.
Virtually all plant parts rotate slightly about a central axis, a movement called circumnutation, but vines show exaggerated circumnutation. This study contrasts circumnutation in two congeneric twining vines, specifically focusing on differences in erect and prostrate shoots, and examines the impact of circumnutation on exploitation of available climbing supports and exploration for more distant ones. Elongating shoots of Lonicera japonica and L. sempervirens growing in a common garden were classed as (1) erect but not climbing, (2) climbing on a trellis, or (3) prostrate, and their circumnutation quantified by tracking the compass direction of shoot tips. To quantify the impact of different circumnutation behaviors, the climbing success of erect shoots and the maximum dispersion and rooting success of prostrate shoots were measured. Erect shoots of both species circumnutated at similar rates (averaging 31°/h), and did not differ in their success rate of exploiting trellises (averaging 76.8%). Prostrate shoots differed, with those produced by L. japonica having reduced circumnutation. In contrast, prostrate shoots of L. sempervirens continued to circumnutate as much as erect shoots. The specialized circumnutation behavior of the prostrate shoots of L. japonica results in increased rooting success and maximum dispersion compared to the unspecialized shoots of L. sempervirens.


Ergometer training volume and previous injury predict back pain in rowing; strategies for injury prevention and rehabilitation.
The most commonly reported injury site in rowers is the lower back. Research in recent years has focused on epidemiology and biomechanical analyses to try and understand mechanisms that contribute to this injury's onset. Injury surveillance mainly comprises retrospective questionnaires and reviews of medical records with a lack of prospective data. Of studies that reported 12-month data, the incidence of low back pain ranged from 31.8 to 51% of the cohort. Of the limited studies that specifically examined low back pain in rowers, (1) history of lumbar spine injury and (2) volume of ergometer training were the most significant risk factors for injury onset. Studies of technique on the rowing ergometer have indicated the importance of lumbopelvic rotation during rowing. Greater pelvic rotation at either end of the stroke is ideal-as opposed to lumbar flexion and extension; this tends to be poorly demonstrated in novice rowers on ergometers. Furthermore, technique can deteriorate with the demands of rowing intensity and duration, which puts the rower returning from injury at additional risk.


Knees with anteromedial osteoarthritis show a substantial phenotypic variation prior and following medial unicompartmental knee arthroplasty.
The primary aim of this study was to evaluate the phenotypic variation using the Coronal Plane Alignment of the Knee (CPAK) classification among 1000 knees with anteromedial osteoarthritis (OA) both prior to and following medial unicompartmental knee arthroplasty (UKA). The secondary aim of this study was to investigate whether knees maintained their preoperative CPAK phenotype and to evaluate the phenotypic alterations following medial UKA. The CPAK classification was used to analyze 1000 knees that underwent medial UKA as treatment for anteromedial OA. Knees were categorized into nine distinct CPAK phenotypes based on their arithmetic hip-knee-ankle angle (aHKA), which estimates the pre-arthritic alignment, and joint line obliquity (JLO), both pre- and postoperatively. Phenotypic variation was analyzed by sex and age, and the phenotypic alterations following medial UKA were evaluated by phenotype. Preoperatively, CPAK phenotype I had the highest prevalence (45.0%). Among males, the preoperative prevalence of CPAK phenotype I was significantly higher compared to females (53.2% vs. 35.0%, respectively; p ≤ .001), whereas females exhibited a significantly higher occurrence of CPAK phenotype V compared to males (9.8% vs. 4.4%, respectively; p ≤ .015). Following medial UKA, CPAK phenotype II had the highest prevalence (53.3%). Overall, 45.1% of knees maintained their preoperative CPAK phenotype following medial UKA, which was most frequently observed among CPAK phenotype II (67.7%) and III (65.8%). There is a substantial variation in CPAK phenotypes among knees with anteromedial OA, as well as following treatment with medial UKA. This variability challenges the assumption of uniform characteristics among knees with an identical wear pattern associated with anteromedial OA and emphasizes the complexity and variability of this specific form of OA. III, Retrospective cohort study.


Non-syndromic multiple supernumerary premolars: Clinicoradiographic report of five cases.
Hyperdontia or supernumerary teeth in both arches without any syndromic manifestation are extremely rare. Supernumerary teeth are commonly associated with Gardner's syndrome, cleft lip and palate, cleidocranial dysplasia and trichorhinopha-langeal syndrome. Five cases of non-syndromic multiple premolars of maxillary and mandibular arches in Indian patients are presented here. This case series reports three cases with multiple (9 in maximum), bilaterally impacted and erupted supernumerary teeth and two cases with supernumerary premolars in non-syndromic cases from Indian patients. Supernumerary teeth can be present in any region of the oral cavity. Although the occurrence of maxillary para-premolars is rare, radio-logical investigations play a major and decisive role in determining the management of such cases.


Laboratory diagnostic techniques for Entamoeba species.
The genus Entamoeba contains many species, six of which (Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Entamoeba polecki, Entamoeba coli, and Entamoeba hartmanni) reside in the human intestinal lumen. Entamoeba histolytica is the causative agent of amebiasis and is considered a leading parasitic cause of death worldwide in humans. Although recent studies highlight the recovery of E. dispar and E. moshkovskii from patients with gastrointestinal symptoms, there is still no convincing evidence of a causal link between the presence of these two species and the symptoms of the host. New approaches to the identification of E. histolytica are based on detection of E. histolytica-specific antigen and DNA in stool and other clinical samples. Several molecular diagnostic tests, including conventional and real-time PCR, have been developed for the detection and differentiation of E. histolytica, E. dispar, and E. moshkovskii in clinical samples. The purpose of this review is to discuss different methods that exist for the identification of E. histolytica, E. dispar, and E. moshkovskii which are available to the clinical diagnostic laboratory. To address the need for a specific diagnostic test for amebiasis, a substantial amount of work has been carried out over the last decade in different parts of the world. The molecular diagnostic tests are increasingly being used for both clinical and research purposes. In order to minimize undue treatment of individuals infected with other species of Entamoeba such as E. dispar and E. moshkovskii, efforts have been made for specific diagnosis of E. histolytica infection and not to treat based simply on the microscopic examination of Entamoeba species in the stool. The incorporation of many new technologies into the diagnostic laboratory will lead to a better understanding of the public health problem and measures to control the disease.


Metallic Burden of Deciduous Teeth and Childhood Behavioral Deficits.
Attention Deficit/Hyperactivity Disorder (ADHD) affects 5%-8% of children in the U.S. (10% of males and 4% of females). The contributions of multiple metal exposures to the childhood behavioral deficits are unclear, although particular metals have been implicated through their neurotoxicity. The objective of this study was to test the hypothesis that the body burden of Mn is positively correlated with ADHD symptoms. We also investigated the putative roles of Ca, Fe, Pb, and Hg. We collected shed molars from 266 children (138 boys and 128 girls) who lost a tooth between 11 and 13 years of age. The molars were analyzed for metals using ICP-OES. The third grade teacher of each child completed the Teacher's Disruptive Behavior Disorders Rating Scale (DBD) to produce a score for "Total Disruptive Behavior" and subscale scores for "Attention Deficit Hyperactivity Disorder", Hyperactivity/Impulsivity, Inattention, and Oppositional/Defiant. The mean Mn, Fe, Pb and Ca concentrations found in teeth was 6.1 ± 5.7 µg/g, 22.7 ± 24.1 µg/g, 0.9 ± 1.4 µg/g, and 6.0 × 105 ± 1.6 × 105 µg/g, respectively. Hg was not detected. No significant association was found between Mn and behavioral deficits. Ca was significantly negatively associated, and Pb showed a significant positive association with Hyperactivity/Impulsivity, Inattention, and Oppositional/Defiant Disorders. These findings call into question the putative independent association of manganese exposure and behavioral deficits in children, when the balance of other metallic burden, particularly Ca and Pb burdens play significant roles.


The influence of travel time on emergency obstetric care seeking behavior in the urban poor of Bangladesh: a GIS study.
Availability of Emergency Obstetric Care (EmOC) is crucial to avert maternal death due to life-threatening complications potentially arising during delivery. Research on the determinants of utilization of EmOC has neglected urban settings, where traffic congestion can pose a significant barrier to the access of EmOC facilities, particularly for the urban poor due to costly and limited transportation options. This study investigates the impact of travel time to EmOC facilities on the utilization of facility-based delivery services among mothers living in urban poor settlements in Sylhet, Bangladesh. A cross-sectional EmOC health-seeking behavior survey from 39 poor urban clusters was geo-spatially linked to a comprehensive geo-referenced dataset of EmOC facility locations. Geo-spatial techniques and logistic regression were then applied to quantify the impact of travel time on place of delivery (EmOC facility or home), while controlling for confounding socio-cultural and economic factors. Increasing travel time to the nearest EmOC facility is found to act as a strong deterrent to seeking care for the urban poor in Sylhet. Logistic regression results indicate that a 5-min increase in travel time to the nearest EmOC facility is associated with a 30 % decrease (0.655 odds ratio, 95 % CI: 0.529-0.811) in the likelihood of delivery at an EmOC facility rather than at home. Moreover, the impact of travel time varies substantially between public, NGO and private facilities. A 5-min increase in travel time from a private EmOC facility is associated with a 32.9 % decrease in the likelihood of delivering at a private facility, while for public and Non-Government Organizations (NGO) EmOC facilities, the impact is lower (28.2 and 28.6 % decrease respectively). Other strong determinants of delivery at an EmOC facility are the use of antenatal care and mother's formal education, while Muslim mothers are found to be more likely to deliver at home. Geospatial evidence points to the need to strengthen referral and emergency transport systems in order to reduce urban travel time, and establish or relocate EmOC facilities closer to where the poor reside. However, female education and antenatal care coverage remain the most important determinants of facility delivery.


[Efficacy and tolerance of a five day course of ciprofloxacin in the treatment of urinary tract infection in women. Multicenter study].
There is no consensus about the ideal antimicrobial agent and duration of treatment for urinary tract infection in women. To assess the efficacy of a five days course of ciprofloxacin for the treatment of urinary tract infection in women. Women with urinary tract infection were treated with ciprofloxacin (Baycip, Bayer) 250 mg bid during 5 days. Patients were evaluated three to four days after treatment start, two to seven days and one month after treatment end. Of 101 eligible women, 96 aged 18 to 65 years old, coming from three major Chilean cities, participated in the study and 80 completed the follow up period. There was a 95% clinical success, 2.5% partial improvement and 2.5% treatment failure. The causal microorganism was erradicated in 90% of cases, in 1.2% treatment failed and in 8.7% a re-infection occurred. Adverse effects attributable to the drug were observed in 12 patients (headache in 3, gastrointestinal disturbances in 8, somnolence in 1 and irritability in 1). Ciprofloxacin is an useful antimicrobial for the treatment of lower urinary tract infection in women.


Cementless total hip arthroplasty with extended sliding trochanteric osteotomy for high congenital hip dislocation: A retrospective study.
Total hip arthroplasty (THA) for high congenital hip dislocation (CHD) is technically demanding. The purpose of this retrospective study was to evaluate the results of cementless THA combined with extended sliding trochanteric osteotomy. We also assessed whether chronic low back pain was relieved after surgery.The study included 19 patients (23 hips) with high CHD treated with cementless THA using extended sliding trochanteric osteotomy technique. Clinical and radiographic outcomes were evaluated.Harris Hip Score, WOMAC score, visual analog scale for low back pain and Trendelenburg sign were significantly improved (P < 0.01) compared with the preoperative. Average limb-length discrepancy in the 15 unilateral hips was reduced from 38.2 ± 7.9 mm to 6.7 ± 4.1 mm (P < 0.01). No dislocation, deep vein thrombosis, or infection occurred. Two patients (8.7%) developed sciatic nerve palsy. One (4.3%) developed symptomatic greater trochanteric bursitis. Two (8.7%) sustained proximal femur shaft fracture during implantation of the femoral component. All femoral components showed successful bony ingrowth at the final follow-up. No stem subsidence was detected. There was no acetabular loosening. Bony union of the reattached greater trochanter was obtained in all hips. Wire breakage occurred in 3 hips (13%).Cementless THA with extended sliding trochanteric osteotomy may be appropriate options for patients with high CHD.


Combined effects of Er:YAG laser and nano-carbonate apatite dentifrice on dentinal tubule occlusion: in vitro study.
The aim of this study was to evaluate the occluding effects of Er:YAG laser and dentifrice containing nano-carbonate apatite (n-CAP) on the dentinal tubules. Although there are various treatment methods for dentin hypersensitivity (DH), more effective methods are still being sought. Sound human root dentin specimens were etched with 6% citric acid to open the dentinal tubules. The specimens were divided randomly into four groups: no treatment (control group), toothbrushing using 20% n-CAP dentifrice (n-CAP group), Er:YAG laser irradiation (LI group), and LI after n-CAP (combined group). The degree of occlusion of the dentinal tubules and the thickness of the deposition layer were evaluated by scanning electron microscopy and an image analyzer. It was shown that the occlusion effect of the n-CAP group was 87%, that of the combined group was 54%, and that of the LI group was 52%, compared with that of the control group. And the n-CAP particles were observed not only under the irradiated surface and but also above the surface in the combined group. This combination method has the potential for enhancement of the dentinal tubule occlusion. However, further study is needed to present the combination effects with additional analysis, and the long-term effects.


The Relationship between Wormlike Micelle Scission Free Energy and Micellar Composition: The Case of Sodium Lauryl Ether Sulfate and Cocamidopropyl Betaine.
The scission energy is the difference in free energy between two hemispherical caps and the cylindrical region of a wormlike micelle. This energy difference determines the logarithm of the average micelle length, which affects several macroscopic properties such as the viscosity of viscoelastic fluids. Here we use a recently published method by Wang et al. ( Langmuir, 2018, 34, 1564-1573) to directly calculate the scission energy of micelles composed of monodisperse sodium lauryl ether sulfate (SLESnEO), an anionic surfactant. Utilizing dissipative particle dynamics (DPD), we perform a systematic study varying the number of ethoxyl groups (n) and salt concentration. The scission energy increases with increasing salt concentration, indicating that the formation of longer micelles is favored. We attribute this to the increased charge screening that reduces the repulsion between head groups. However, the scission energy decreases with increasing number of ethoxyl groups as the flexibility of the head group increases and the sodium ion becomes less tightly bound to the head group. We then extend the analysis to look at the effect of a common cosurfactant, cocamidopropyl betaine (CAPB), and find that its addition stabilizes wormlike micelles at a lower salt concentration.


Immunohistochemical localization of zona pellucida proteins ZPA, ZPB and ZPC in human, cynomolgus monkey and mouse ovaries.
The zona pellucida of mammalian oocytes plays an important role in binding and activation of sperm cells during the molecular events leading to fertilization. The genes coding for the three zona pellucida glycoproteins ZPA, ZPB, and ZPC of various species including mouse, dog, and human have been cloned and sequenced by several groups. However, it has remained a matter of debate as to whether the oocytes alone or in conjunction with the surrounding granulosa cells express and deposit these proteins to form the zona pellucida matrix. Addressing this unresolved issue, we assessed the expression and localization of all three zona pellucida proteins in ovaries of human, cynomolgus monkey and mice using immunohistochemical methods. In addition, oocyte-specific expression of ZPC from the primordial stage onward was confirmed by in situ hybridization. In sections of human ovaries, ZPA, ZPB, and ZPC proteins were immunohistochemically detected in the cytoplasms of primordial oocytes and during later stages of folliculogenesis in the zona pellucida matrices of oocytes. In sections fixed with formalin, a clear homogeneous ring was visible around the oocyte and no staining of granulosa cells was observed. In contrast, staining of ZP proteins was also observed between granulosa cells when Bouin's reagent had been used for tissue fixation. Thus, the original zona pellucida architecture was better preserved by formalin fixation. We further demonstrated that dissolution of the zona pellucida of isolated bovine oocytes occurred after they were exposed to Bouin's reagent. In summary, these results demonstrate that in mice, monkeys and humans, zona proteins are expressed and assembled exclusively by the oocyte and not by the granulosa cells. Previously observed results of ZP expression by an involvement of granulosa cells might therefore be the result of an improper fixation of the tissues leading to the disruption of the zona pellucida. Additionally this study highlights the importance of choosing the correct fixative for immunohistochemistry, not only for the usual reason of retaining antigenicity, but rather to retain the entire architectural structure.


[Efficacy of PGF2 alpha on milk ejection during the sexual cycle of the cow].
After 30 days of lactation, 15 non-pregnant French Friesian cows were divided into groups A and B, balanced by the milk yield and lactation number of the cows. The 7 cows of group A were given a series of intrajugular prostaglandin F2 alpha (Upjohn Dinolytic) injections every morning, administered in increasing logarithmic doses (1, 4, 16, 64, 256, 1 024 micrograms). The aim of the experiment was to determine the minimal dose that would induce an increase in intramammary pressure (IMP) during the sexual cycle and milk let-down through a cannula in the teat cup. The 8 cows of group B were given a single injection of 256 micrograms of PGF2 alpha every morning to determine the changes during the sexual cycle in the response parameters of IMP (latency and amplitude) or of milk let-down (latency and collected volume) through a cannula. The following results were obtained: 1. The minimal dose of prostaglandin inducing milk let-down varied during the cycle; it was lower (1 to 16 micrograms) during the luteal phase (D4 to D12) than during the perioestral phase (256 to greater than 1 024 micrograms from D - 2 to D + 3) (figs. 2, 3). 2. During daily administration of the same dose (256 micrograms) of PGF2 alpha no response was obtained during the perioestral phase, while during the luteal phase; - maximal IMP deflection reached 12 to 14 mmHg (figs. 5, 6); - the volume of milk ejected was on an average higher than 3 liters per quarter (figs. 5, 6); - the latency time between injection and let down response was about 1.8 times shorter between D10 and D12 than between D4 and D5 (fig. 7). 3. The cyclic changes in the milk ejection parameters caused by PGF2 alpha were closely related to the plasma progesterone level (figs. 2, 3, 5, 6). The coefficients of correlation between the progesterone level and the - threshold dose: r = - 0.809** (group A), - volume of ejected milk: r = + 0.872*** (group B), - deflection amplitude: r = + 0.805*** (group B) were always significantly higher than the threshold of 0.001. Various hypotheses concerning the endocrine control of cyclic variations in milk let-down under the effect of PGF2 alpha are discussed.


Lessons from unified Germany and their implications for healthcare in the unification of the Korean Peninsula.
This study investigated the German experience in the transition to a unified health care system and suggests the following implications for Korea. First, Germany could have made use of the unification process better if there had been a good road map. Therefore Korea must develop a well prepared road map that considers all possible situations. Second, Germany saw an opportunity for the improvement of the health care system in the early stage of unification but could not take advantage of it because the situation changed dramatically and they had not sufficiently prepared for it. Korea should take into account the opportunity for improvement of the present health care system, such as the roles of public health and traditional medicine. Thirdly, the conditions of North Korea seem to be far worse than those of former East Germany and also worse than even those of other transition countries [corrected].Therefore Korea should design a long-term road map taking as many variables into account as possible, including the different rigid way of thinking and the interrelationship among the social sectors. Fourthly, during the German reunification unexpected factors changed the direction of the events. Korea should have a separate plan for the unexpected factors.


Downregulation of miR-23a and miR-27a following experimental traumatic brain injury induces neuronal cell death through activation of proapoptotic Bcl-2 proteins.
MicroRNAs (miRs) are small noncoding RNAs that negatively regulate gene expression at the post-transcriptional level. To identify miRs that may regulate neuronal cell death after experimental traumatic brain injury (TBI), we profiled miR expression changes during the first several days after controlled cortical impact (CCI) in mice. miR-23a and miR-27a were rapidly downregulated in the injured cortex in the first hour after TBI. These changes coincided with increased expression of the proapoptotic Bcl-2 family members Noxa, Puma, and Bax. In an etoposide-induced in vitro model of apoptosis in primary cortical neurons, miR-23a and miR-27a were markedly downregulated as early as 1 h after exposure, before the upregulation of proapoptotic Bcl-2 family molecules. Administration of miR-23a and miR-27a mimics attenuated etoposide-induced changes in Noxa, Puma, and Bax, reduced downstream markers of caspase-dependent (cytochrome c release and caspase activation) and caspase-independent (apoptosis-inducing factor release) pathways, and limited neuronal cell death. In contrast, miRs hairpin inhibitors enhanced etoposide-induced neuronal apoptosis and caspase activation. Importantly, administration of miR-23a and miR-27a mimics significantly reduced activation of Puma, Noxa, and Bax as well as attenuated markers of caspase-dependent and -independent apoptosis after TBI. Furthermore, miR-23a and miR-27a mimics significantly attenuated cortical lesion volume and neuronal cell loss in the hippocampus after TBI. These findings indicate that post-traumatic decreases in miR-23a and miR-27a contribute to neuronal cell death after TBI by upregulating proapoptotic Bcl-2 family members, thus providing a novel therapeutic target.


Support and negation of colorectal cancer risk prevention behaviors: analysis of spousal discussions.
The shared social context created in a marriage may be important in motivating engagement in health behaviors, but spousal influence may not be uniformly applied. Our goal was to examine how spouses discuss health behaviors relevant for colorectal cancer (CRC) risk-reduction to better understand how spouses exert or fail to exert influence. In this pilot study, first degree relatives of CRC patients and their spouses completed demographic and self-reported health questionnaires. After a genetic counseling session regarding risk and risk reduction, couples engaged in a semi-structured discussion task to discuss lifestyle choices they currently undertake or could undertake to reduce risk. Demographic and health data was analyzed using descriptive statistics. Using a directed content analysis based on the transtheoretical model of behavior change, we coded discussion transcripts for depth and direction of talk for seven behavioral CRC risk factors. Spouses engaged in several strategies to reduce their risk for CRC, and problem-solved together to increase these preventative efforts. All couples mentioned diet and exercise as important factors in CRC risk; however, other risk factors received less attention. Despite evidence of support and encouragement, spouses ignored, minimized, or negated the importance of some health behaviors. Spousal influence could be an important tool to improve participation in health behaviors, but more guidance may be necessary to hold couples accountable to evidence-based guidelines to reduce risk. Health care providers should address couples as a unit to assess and address health behaviors.


Investigation of lignocellulolytic enzymes during different growth phases of Ganoderma lucidum strain G0119 using genomic, transcriptomic and secretomic analyses.
Ganoderma lucidum is a medicinal mushroom that is well known for its ability to enhance human health, and products made from this fungus have been highly profitable. The substrate-degrading ability of G. lucidum could be related to its growth. CAZy proteins were more abundant in its genome than in the other white rot fungi models. Among these CAZy proteins, changes in lignocellulolytic enzymes during growth have not been well studied. Using genomic, transcriptomic and secretomic analyses, this study focuses on the lignocellulolytic enzymes of G. lucidum strain G0119 to determine which of these degradative enzymes contribute to its growth. From the genome sequencing data, genes belonging to CAZy protein families, especially genes involved in lignocellulose degradation, were investigated. The gene expression, protein abundance and enzymatic activity of lignocellulolytic enzymes in mycelia over a growth cycle were analysed. The overall expression cellulase was higher than that of hemicellulase and lignin-modifying enzymes, particularly during the development of fruiting bodies. The cellulase and hemicellulase abundances and activities increased after the fruiting bodies matured, when basidiospores were produced in massive quantities till the end of the growth cycle. Additionally, the protein abundances of the lignin-modifying enzymes and the expression of their corresponding genes, including laccases and lignin-degrading heme peroxidases, were highest when the mycelia fully spread in the compost bag. Type I cellobiohydrolase was observed to be the most abundant extracellular lignocellulolytic enzyme produced by the G. lucidum strain G0119. The AA2 family haem peroxidases were the dominant lignin-modifying enzyme expressed during the mycelial growth phase, and several laccases might play roles during the formation of the primordium. This study provides insight into the changes in the lignocellulose degradation ability of G. lucidum during its growth and will facilitate the discovery of new approaches to accelerate the growth of G. lucidum in culture.


On the discrimination of multiple phytoplankton groups from light absorption spectra of assemblages with mixed taxonomic composition and variable light conditions.
According to recommendations of the international community of phytoplankton functional type algorithm developers, a set of experiments on marine algal cultures was conducted to (1) investigate uncertainties and limits in phytoplankton group discrimination from hyperspectral light absorption properties of assemblages with mixed taxonomic composition, and (2) evaluate the extent to which modifications of the absorption spectral features due to variable light conditions affect the optical discrimination of phytoplankton. Results showed that spectral absorption signatures of multiple species can be extracted from mixed assemblages, even at low relative contributions. Errors in retrieved pigment abundances are, however, influenced by the co-occurrence of species with similar spectral features. Plasticity of absorption spectra due to changes in light conditions weakly affects interspecific differences, with errors <21% for retrievals of pigment concentrations from mixed assemblages.


Active-space N-representability constraints for variational two-particle reduced density matrix calculations.
The ground-state energy of a system of fermions can be calculated by minimizing a linear functional of the two-particle reduced density matrix (2-RDM) if an accurate set of N-representability conditions is applied. In this Letter we introduce a class of linear N-representability conditions based on exact calculations on a reduced active space. Unlike wave-function-based approaches, the 2-RDM methodology allows us to combine information from calculations on different active spaces. By adding active-space constraints, we can iteratively improve our estimate for the ground-state energy. Applying our methodology to a 1D Hubbard model yields a significant improvement over traditional 2-positivity constraints with the same computational scaling.


[Modification of Escherichia coli RNA polymerase by diethylpyrocarbonate. II. Binding and unwinding of double-stranded DNA].
E. coli DNA dependent RNA polymerase was modified by diethylpyrocarbonate. Binding to a double-stranded DNA and unwinding of the DNA at the enzyme binding site by the modified enzyme were examined. It was found that RNA polymerase reversibly lost the ability to unwind DNA helix as well as the RNA synthetic activity when 9 to 11 histidyl residues of the enzyme were modified. In addition ot modification of the most reactive sulfhydryl or amino groups of the enzyme accompanying histidyl residues modification results in irreversible decrease of the salt concentration which is necessary to remove the enzyme from DNA cellulose column. Further modification of the less reactive sulfhydryl or amino groups leads to irreversible loss of the DNA binding ability and to the enzyme structure alteration.


Mechanisms of nitrogen dioxide toxicity in humans.
These studies were undertaken to evaluate short-term respiratory effects and identify markers of nitrogen dioxide toxicity during exposures designed to approximate realistic conditions. With the development of bronchoalveolar lavage as a clinical investigative technique, the evaluation focused on the assessment of effects induced at the alveolar level. The exposure protocols were designed to assess the duration of nitrogen dioxide-induced effects and determine exposure-response relationships. Groups of normal, nonsmoking volunteers of both sexes between the ages of 18 and 40 years, without airway hyperreactivity, constituted the study population. The exposure protocols required a total of three to five days for each subject, depending on the timing of bronchoalveolar lavage. Subjects were exposed to nitrogen dioxide or air for three hours in a double-blind, randomized fashion in a 45-m3 environmental chamber, with intermittent exercise sufficient to quadruple minute ventilation. Pulmonary function was measured during and after exposure, and airway reactivity to carbachol was assessed before and after exposure. Lavaged cells were examined for their capacity to inactivate influenza virus and secrete IL-1 in vitro. Cell-free lavage fluid was analyzed for total protein, albumin, alpha 2-macroglobulin, arylsulfatase, and alpha 1-protease inhibitor. The studies were undertaken in three phases, each of approximately one year's duration. In Phase 1, 15 subjects were exposed to a background concentration of 0.05 parts per million2 (ppm) nitrogen dioxide and to three 15-minute peaks of 2.0 ppm, and underwent bronchoalveolar lavage 3.5 hours after nitrogen dioxide exposure. During Phase 2, 8 subjects were exposed to continuous 0.60 ppm nitrogen dioxide and underwent bronchoalveolar lavage 18 hours later. Finally, in Phase 3, 15 subjects were exposed to continuous 1.5 ppm nitrogen dioxide and underwent bronchoalveolar lavage 3.5 hours after exposure. No significant symptomatic or pulmonary function changes could be detected in response to any of the nitrogen dioxide exposures. However, a small but significant increase in airway reactivity was observed in normal subjects after exposure to 1.5 ppm nitrogen dioxide. Following the highest dose of carbachol (10 mg/mL), the forced expiratory volume in one second decreased 7.5 +/- 1.1 percent after nitrogen dioxide exposure compared to 4.8 +/- 1.1 percent after exposure to air (p less than 0.05). No symptoms were induced in any of the groups by the carbachol exposures. Analyses of cells recovered by bronchoalveolar lavage during all three phases revealed no differences in total cell recovery, cell viability, or differential cell counts.(ABSTRACT TRUNCATED AT 400 WORDS)


Proliferative and apoptotic responses in cancers with special reference to oral cancer.
The study of signal transduction pathways for mechanisms of apoptosis and proliferation has significantly advanced our understanding of human cancer, subsequently leading to more effective treatments. Discoveries of growth factors and oncogenes, especially those that function through phosphorylation on tyrosine residues, have greatly benefited our appreciation of the biology of cancer. The regulation of proliferation and apoptosis through phosphorylation via tyrosine kinases and phosphatases is discussed, as well as the contributions of other systems, such as serine and threonine kinases and phosphatases. Receptors with seven-transmembrane domains, steroid hormones, genes, and "death domains" will also be discussed. This review attempts to compare the regulation of the growth of normal tissues and cancers with an effort to highlight the current knowledge of these factors in the growth regulation of oral/oropharyngeal cancers. Despite the strides made in our understanding of growth regulation in human cancers, the study of oral/oropharyngeal cancer specifically lags behind. More research must be done to further our understanding of oral cancer biology, if we are to develop better, more effective treatment protocols.


Top-gated chemical vapor deposition grown graphene transistors with current saturation.
Graphene transistors are of considerable interest for radio frequency (rf) applications. In general, transistors with large transconductance and drain current saturation are desirable for rf performance, which is however nontrivial to achieve in graphene transistors. Here we report high-performance top-gated graphene transistors based on chemical vapor deposition (CVD) grown graphene with large transconductance and drain current saturation. The graphene transistors were fabricated with evaporated high dielectric constant material (HfO(2)) as the top-gate dielectrics. Length scaling studies of the transistors with channel length from 5.6 μm to 100 nm show that complete current saturation can be achieved in 5.6 μm devices and the saturation characteristics degrade as the channel length shrinks down to the 100-300 nm regime. The drain current saturation was primarily attributed to drain bias induced shift of the Dirac points. With the selective deposition of HfO(2) gate dielectrics, we have further demonstrated a simple scheme to realize a 300 nm channel length graphene transistors with self-aligned source-drain electrodes to achieve the highest transconductance of 250 μS/μm reported in CVD graphene to date.


Genetic analysis of type O viruses responsible for epidemics of foot-and-mouth disease in North Africa.
The nucleotide sequences of the 3' end of the capsid-coding region were determined for 30 serotype O foot-and-mouth disease (FMD) viruses isolated between 1987 and 1994 from outbreaks in North Africa and the Middle East. These sequences were compared with the previously published sequences of 9 field virus isolates from the Middle East and 5 vaccine virus strains, 3 of which originated from the Middle East (O1/Turkey/Manisa/69, O1/Sharquia/Egypt/72 and O1/Israel/2/85) and 2 from Europe (O1/Lausanne/Switzerland/65 and O2/Brescia/Italy/47). Cluster analysis of these sequences using the unweighted pair group mean average (UPGMA) method showed: (i) that the FMD viruses isolated from North Africa and the Middle East were very different from the classical European vaccine strains; (ii) that all the viruses isolated during the 1989-92 North African epidemic formed a cluster differing by no more than 6% from each other; (iii) a virus isolated in Libya in 1988 was unrelated to the aforementioned epidemic; and (iv) viruses from a second, less extensive epidemic, occurring in 1994, fell into yet another cluster.


Transurethral Microwave Thermotherapy for Benign Prostatic Hyperplasia: An Updated Cochrane Review.
To assess the effects of transurethral microwave thermotherapy (TUMT) for the treatment of lower urinary tract symptoms in men with benign prostatic hyperplasia (BPH). We performed a comprehensive search using multiple databases up to May 2021, with no language or publication status restrictions. We included parallel-group randomized controlled trials of participants with BPH who underwent TUMT. We used standard Cochrane methods, including a GRADE assessment of the certainty of the evidence (CoE). In this update of a previous Cochrane review, we included 16 trials with 1,919 participants. TUMT probably results in little to no difference in urologic symptom scores at short-term follow-up compared to transurethral resection of the prostate (TURP). There is likely to be little to no difference in the quality of life. TUMT likely results in fewer major adverse events. TUMT, however, probably results in a large increase in the need for retreatment. There may be little to no difference in erectile function between these interventions. However, TUMT may result in fewer cases of ejaculatory dysfunction compared to TURP. The overall CoE was moderate to low. TUMT provides a similar reduction in urinary symptoms compared to TURP, with fewer major adverse events and fewer cases of ejaculatory dysfunction at short-term follow-up. However, TUMT probably results in a large increase in retreatment rates. Study limitations and imprecision reduced the confidence we can place in these results.


Surgical therapy attenuates abdominal and extra-abdominal inflammation in experimental peritonitis.
This study examines the influence of surgical management (elimination of the infectious focus and abdominal lavage) on survival and the inflammatory response in the various compartments of the body: local (abdomen), systemic (blood) and distant organ (lungs). Peritonitis was established in mice by cecal ligation and puncture (CLP). After 24 h, a group was made in which the infected cecum was resected and the abdominal cavity was lavaged (RES), and another group that received no surgical resection (NoRES). Survival was examined over a period of 96 h. Mice were sacrificed at 24 (sham and CLP), 48 and 72 h after CLP to measure inflammatory parameters. Survival was significantly lower is NoRES compared to sham and RES (p = 0.006, p = 0.014, respectively). Intraperitoneal parameters were improved in the RES group compared to sham but results were not significantly different between groups. In plasma, levels of interleukin-6 (IL-6) were decreased in RES (p = 0.048). Accordingly, anti-inflammatory IL-10 in plasma was increased in this group (p = 0.031). In the lung, keratinocyte-derived chemokine (KC) and myeloperoxidase (MPO) was reduced indicating decreased granulocytes accumulation in the lung in the RES group (p = 0.012 and p = 0.004, respectively). In experimental secondary peritonitis surgical management improves survival and attenuates the inflammatory response predominantly in the extra-abdominal compartments. This illustrates the importance of surgery in prevention of multiple organ failure and denotes the compartmentalized character of the inflammatory response. This polymicrobial model with implicated surgical intervention reflects the clinical situation and may be more appropriate to test therapeutic interventions than a model involving only CLP.


Development of Phasmarhabditis hermaphrodita (and members of the Phasmarhabditis genus) as new genetic model nematodes to study the genetic basis of parasitism.
The genetic mechanisms of how free-living nematodes evolved into parasites are unknown. Current genetic model nematodes (e.g. Caenorhabditis elegans) are not well suited to provide the answer, and mammalian parasites are expensive and logistically difficult to maintain. Here we propose the terrestrial gastropod parasite Phasmarhabditis hermaphrodita as a new alternative to study the evolution of parasitism, and outline the methodology of how to keep P. hermaphrodita in the lab for genetic experiments. We show that P. hermaphrodita (and several other Phasmarhabditis species) are easy to isolate and identify from slugs and snails from around the UK. We outline how to make isogenic lines using 'semi-natural' conditions to reduce in-lab evolution, and how to optimize growth using nematode growth media (NGM) agar and naturally isolated bacteria. We show that P. hermaphrodita is amenable to forward genetics and that unc and sma mutants can be generated using formaldehyde mutagenesis. We also detail the procedures needed to carry out genetic crosses. Furthermore, we show natural variation within our Phasmarhabditis collection, with isolates displaying differences in survival when exposed to high temperatures and pH, which facilitates micro and macro evolutionary studies. In summary, we believe that this genetically amenable parasite that shares many attributes with C. elegans as well as being in Clade 5, which contains many animal, plant and arthropod parasites, could be an excellent model to understand the genetic basis of parasitism in the Nematoda.


Structural insights into microneme protein assembly reveal a new mode of EGF domain recognition.
The obligate intracellular parasite Toxoplasma gondii, a member of the phylum Apicomplexa that includes Plasmodium spp., is one of the most widespread parasites and the causative agent of toxoplasmosis. Adhesive complexes composed of microneme proteins (MICs) are secreted onto the parasite surface from intracellular stores and fulfil crucial roles in host-cell recognition, attachment and penetration. Here, we report the high-resolution solution structure of a complex between two crucial MICs, TgMIC6 and TgMIC1. Furthermore, we identify two analogous interaction sites within separate epidermal growth factor-like (EGF) domains of TgMIC6-EGF2 and EGF3-and confirm that both interactions are functional for the recognition of host cell receptor in the parasite, using immunofluorescence and invasion assays. The nature of this new mode of recognition of the EGF domain and its abundance in apicomplexan surface proteins suggest a more generalized means of constructing functional assemblies by using EGF domains with highly specific receptor-binding properties.


Potential microbiological contamination of effluents in poultry and swine abattoirs.
Health risks in the effluents of seven swine abattoirs and of seven poultry abattoirs were evaluated with regard to environment degradation and to dissemination of pathogenic microorganisms during the rainy and dry seasons. Supply-water samples from affluents and effluents of the treatment systems at different sites within the abattoir processing system were analysed. Similarly, water samples from the three recipient sites (emission point, 100 m upstream, 100 m downstream) were also analysed. Temperature, free residual chlorine (FRC), total coliform bacteria, Escherichia coli, enterococci, identification and serotyping of salmonellae were assessed. Scalding is the most significant stage in the slaughtering chain (P<0.05) when temperature is taken into account. Temperatures at effluents and at the sampled sites in the water bodies accorded to state and federal legislation standards. Supply waters did not meet the standards for FRC and microbial count standards according to the Ministry of Health and within limits imposed by the Industrial and Sanitary Inspection Regulations for Animal Products. Feather plucking and evisceration in poultry slaughter and the cleansing of carcasses and facilities in poultry and swine slaughtering had the highest contamination impact. The three loci at the water bodies were above the microbiological standards for classes II and III sites, in conformity with Law 8468 of the state of São Paulo, Brazil and Conama. Salmonella was found at several sites during slaughter, at both types of abattoirs, including in the effluent treatment system. This showed that these sites were the dissemination sources of the microorganism.


Membrane potential of CA3 hippocampal pyramidal cells during postnatal development.
A depolarized resting membrane potential has long been considered to be a universal feature of immature neurons. Despite the physiological importance, the underlying mechanisms of this developmental phenomenon are poorly understood. Using perforated-patch, whole cell, and cell-attached recordings, we measured the membrane potential in CA3 pyramidal cells in hippocampal slices from postnatal rats. With gramicidin perforated-patch recordings, membrane potential was -44 +/- 4 (SE) mV at postnatal days P0-P2, and it progressively shifted to -67 +/- 2 mV at P13-15. A similar developmental change of the membrane potential has been also observed with conventional whole cell recordings. However, the value of the membrane potential deduced from the reversal potential of N-methyl-d-aspartate channels in cell-attached recordings did not change with age and was -77 +/- 2 mV at P2 and -77 +/- 2 mV at P13-14. The membrane potential measured using whole cell recordings correlated with seal and input resistance, being most depolarized in neurons with high, several gigaohms, input resistance and low seal resistance. Simulations revealed that depolarized values of the membrane potential in whole cell and perforated-patch recordings could be explained by a shunt through the seal contact between the pipette and membrane. Thus the membrane potential of CA3 pyramidal cells appears to be strongly negative at birth and does not change during postnatal development.


Finding Liebig's law of the minimum.
Liebig's law of the minimum (LLM) is often used to interpret empirical biological growth data and model multiple substrates co-limited growth. However, its mechanistic foundation is rarely discussed, even though its validity has been questioned since its introduction in the 1820s. Here we first show that LLM is a crude approximation of the law of mass action, the state of art theory of biochemical reactions, and the LLM model is less accurate than two other approximations of the law of mass action: the synthesizing unit model and the additive model. We corroborate this conclusion using empirical data sets of algae and plants grown under two co-limiting substrates. Based on our analysis, we show that when growth is modeled directly as a function of substrate uptake, the LLM model improperly restricts the organism to be of fixed elemental stoichiometry, making it incapable of consistently resolving biological adaptation, ecological evolution, and community assembly. When growth is modeled as a function of the cellular nutrient quota, the LLM model may obtain good results at the risk of incorrect model parameters as compared to those inferred from the more accurate synthesizing unit model. However, biogeochemical models that implement these three formulations are needed to evaluate which formulation is acceptably accurate and their impacts on predicted long-term ecosystem dynamics. In particular, studies are needed that explore the extent to which parameter calibration can rescue model performance when the mechanistic representation of a biogeochemical process is known to be deficient.


Elderly patients with COPD require more health care than elderly heart failure patients do in a hospital-based home care setting.
Elderly patients with advanced stages of COPD or chronic heart failure (CHF) often require hospitalization due to exacerbations. We hypothesized that telemonitoring supported by hospital-based home care (HBHC) would detect exacerbations early, thus, reducing the number of hospitalization. We also speculated that patients with advanced COPD or CHF would present differences regarding exacerbation frequency and the need of HBHC. The Health Diary system, based on digital pen technology, was employed. Patients aged ≥65 years with ≥2 hospitalizations the previous year were included. Exacerbations were categorized and treated as either COPD or CHF exacerbation by an experienced physician. All HBHC contacts (home visits or telephone consultations) were registered. Ninety-four patients with advanced diseases were enrolled (36 COPD and 58 CHF subjects) of which 53 subjects (19 COPD and 34 CHF subjects) completed the 1-year study period. Death was the major reason for not finalizing the study. Compared to the 1-year prior inclusion, the intervention significantly reduced hospitalization. Although COPD subjects were younger with less comorbidity, exacerbations and HBHC contacts were significantly greater in this group. COPD subjects exhibit exacerbations more frequently, mainly due to disease characteristics, thus, demanding much more HBHC.


Deficient brainstem encoding of pitch in children with Autism Spectrum Disorders.
Deficient prosody is a hallmark of the pragmatic (socially contextualized) language impairment in Autism Spectrum Disorders (ASD). Prosody communicates emotion and intention and is conveyed through acoustic cues such as pitch contour. Thus, the objective of this study was to examine the subcortical representations of prosodic speech in children with ASD. Using passively evoked brainstem responses to speech syllables with descending and ascending pitch contours, we examined sensory encoding of pitch in children with ASD who had normal intelligence and hearing and were age-matched with typically developing (TD) control children. We found that some children on the autism spectrum show deficient pitch tracking (evidenced by increased Frequency and Slope Errors and reduced phase locking) compared with TD children. This is the first demonstration of subcortical involvement in prosody encoding deficits in this population of children. Our findings may have implications for diagnostic and remediation strategies in a subset of children with ASD and open up an avenue for future investigations.


Clinical research careers: reports from a NHLBI pediatric heart network clinical research skills development conference.
Wyman W. Lai, MD, MPH, and Victoria L. Vetter, MD, MPH. The Pediatric Heart Network (PHN), funded under the U.S. National Institutes of Health-National Heart, Lung, and Blood Institute (NIH-NHLBI), includes two Clinical Research Skills Development (CRSD) Cores, which were awarded to The Children's Hospital of Philadelphia and to the Morgan Stanley Children's Hospital of New York-Presbyterian. To provide information on how to develop a clinical research career to a larger number of potential young investigators in pediatric cardiology, the directors of these two CRSD Cores jointly organized a one-day seminar for fellows and junior faculty from all of the PHN Core sites. The participants included faculty members from the PHN and the NHLBI. The day-long seminar was held on April 29, 2009, at the NHLBI site, immediately preceding the PHN Steering Committee meeting in Bethesda, MD. the goals of the seminar were 1) to provide fellows and early investigators with basic skills in clinical research 2) to provide a forum for discussion of important research career choices 3) to introduce attendees to each other and to established clinical researchers in pediatric cardiology, and 4) to publish a commentary on the future of clinical research in pediatric cardiology. the following chapters are compilations of the talks given at the 2009 PHN Clinical Research Skills Development Seminar, published to share the information provided with a broader audience of those interested in learning how to develop a clinical research career in pediatric cardiology. The discussions of types of clinical research, research skills, career development strategies, funding, and career management are applicable to research careers in other areas of clinical medicine as well. the aim of this compilation is to stimulate those who might be interested in the research career options available to investigators.


Adolescent girls' attitudes toward female genital mutilation: a study in seven African countries.
Background: The study's aim is to examine adolescent girls' attitudes toward the continuation or discontinuation of female genital mutilation (FGM) in association with their demographics in seven different countries in Africa. Methods: Data from the women's survey of the Demographic and Health Surveys (DHS) conducted by the respective ministries (of Health and Family Welfare) in Egypt, Guinea, Kenya, Mali, Niger, Senegal and Sierra Leone were used. Adolescent girls (15-19 years) were included in the current analysis: Egypt (N=636), Guinea (N=1994), Kenya (N= 1767), Mali (N=2791), Niger (N=1835), Senegal (N=3604), Sierra Leone (N=1237). Results: Prevalence of supporting the continuation of FGM among adolescent girls was in Egypt 58%, Guinea 63%, Kenya 16%, Mali 72%, Niger 3%, Senegal 23%, and Sierra Leone 52%. Being Muslim and having low economic status were significantly associated with supporting the continuation of FGM in five of the participating countries. Girls having no education or only primary education in Guinea, Kenya, Mali and Sierra Leone exhibited a higher likelihood of supporting FGM than girls with secondary or higher education. In Egypt, Niger and Senegal there was no association between education and supporting FGM. The girls who stated that they had no exposure to media showed the higher likelihood of supporting FGM in Guinea, Kenya, and Senegal than those with exposure to media. Conclusions: The current study argues that increasing media coverage and education, and reducing poverty are of importance for shifting adolescent girls' attitudes in favor of discontinuation of FGM.


Setting Better-Informed Climate Targets for New Zealand: The Influence of Value and Modeling Choices.
Quantifying greenhouse gas (GHG) emissions and setting GHG emissions budgets for anthropogenic systems are influenced by several value and modeling choices. This study, for the first time, quantified the influence of choice of GHG accounting approach, GHG metric, time horizon, climate threshold, global emissions budget calculation method, and effort-sharing approach, taking New Zealand (NZ) as a case study. First, NZ's production- and consumption-based emissions were quantified using multiregional input-output analysis and applying different GHG metrics (global warming and temperature potentials) and time horizons (20 and 100 years). Second, global emissions budgets for 1.5 °C, 2 °C, and 1 W m-2 climate thresholds were estimated. Budget shares were then assigned to NZ using two effort-sharing approaches (grandfathering and economic value), and emissions were benchmarked against the assigned shares. Finally, the analysis was undertaken at the NZ sector level. The results showed that, for each GHG accounting approach, NZ's total emissions exceeded their budget shares, irrespective of the choices; the largest source of uncertainty was the choice of global emissions budget calculation method, followed by GHG metric, climate threshold, effort-sharing approach, and reference year for the grandfathering approach. The sector-level analysis showed that, while most sectors exceeded their budget shares, some performed within them. The ranking of uncertainty sources was quite different at the sector level, with the choice of effort-sharing approach providing the largest source of uncertainty. Overall, the study indicates the importance of handling value and modeling choices in a transparent way when quantifying GHG emissions and setting emissions budgets for anthropogenic systems.


Quantum material accompanied nonenzymatic cascade amplification for ultrasensitive photoelectrochemical DNA sensing.
A novel cascade photoelectrochemical (PEC) signal amplification sensing strategy was designed and applied in target DNA detection by introducing quantum dots (QD) as the accompanying tag. The CdTe QD was labeled onto a designed sequence of single stranded DNA forming S-QD, which could be further hybridized with another magnetic bead labeled conjugate, leading to a metastable structure. In the presence of the detecting target, the S-QD could be released from the metastable structure via toehold strand displacement. With the help of the designed strand (H), the target could be released and triggered the cascade amplification. In the magnetic field, the S-QD could be easily separated from the reaction mixture and coated on the F-doped tin oxide (FTO) electrode leading to the "signal on" PEC response. The designed cascade amplification photoelectrochemical strategy exhibited a detection limit of 0.76 pM target DNA, and a linear range from 1 pM to 50 nM, indicating promising application in DNA or related analysis. Moreover, compared with other tags such as fluorescent dye, the quantum material leads to less unspecific adsorption and better sensing reproducibility, due to the appropriate nano size and high quantum efficiency.


The assessment of affective disorders in children and adolescents by semistructured interview. Test-retest reliability of the schedule for affective disorders and schizophrenia for school-age children, present episode version.
The reliability of assessment of Research Diagnostic Criteria and DSM-III axis I affective disorders in children and adolescents was studied using a semistructured diagnostic interview. The Schedule for Affective Disorders and Schizophrenia (SADS) for School-Age Children (Kiddie SADS) Present Episode Version, an adaptation of the adult SADS for children was used. Fifty-two subjects, aged 6 through 17 years, were interviewed in a test-retest format by one of three pairs of interviewers. Assessment of symptoms and composite scales of the depressive syndrome were determined to have acceptable reliability, as were three depressive diagnoses. Conduct disorder was assessed with high reliability. Four anxiety disorders and their composite symptoms were assessed with unacceptable reliability; only separation anxiety was assessed with acceptable reliability. The results of this study showed generally lower reliability of symptoms, scales, and diagnoses than did two studies of adults using the SADS.


Selective and Sensitive ZnO Quantum Dots Based Fluorescent Biosensor for Detection of Cysteine.
In the present article, a novel and effective ZnO quantum dots-based fluorescent probe has been developed for the detection of cysteine in different solutions. Firstly, melamine-based fluorescent pre-probe was successfully synthesized via condensation reaction and, then ZnO quantum dots (QDs) were homogenously dispersed into this solution. This fluorescent probe was used for the detection of cysteine in different solutions such as bovine serum albumin and tap water. ZnO QDs were characterized using XRD, nano-particle size analyzer, and FE-SEM techniques. The size of the ZnO QDs was calculated as 28.03±9.86 nm, and 31.95±10.02 nm from Scherrer's equation and nano-particle size analyzer, respectively. The developed fluorescent probe was exhibited a highly selective and sensitive response to the detection of cysteine. Also, the proposed fluorescent probe has a larger Stokes shift value (236 nm). The limit of detection and linear range of ZnO QDs-based fluorescent biosensor were found as 0.642 μM and 0.1-600 μM, respectively. ZnO quantum dot-based fluorescent sensor for L-cysteine.


Bone Turnover Markers Are Not Associated With Hip Fracture Risk: A Case-Control Study in the Women's Health Initiative.
Current guidelines recommend that serum C-terminal telopeptide of type I collagen (CTX) and serum procollagen type 1 aminoterminal propeptide (PINP), measured by standardized assays, be used as reference markers in observational and interventional studies. However, there are limited data to determine whether serum CTX and PINP are associated with hip fracture risk among postmenopausal women. We determined the associations of serum CTX and serum PINP with hip fracture risk among postmenopausal women aged 50 to 79 years at baseline. We performed a prospective case-control study (400 cases, 400 controls) nested in the Women's Health Initiative Observational Study, which enrolled participants at 40 US clinical centers. Cases were women with incident hip fracture not taking osteoporosis medication; hip fractures were confirmed using medical records. Untreated controls were matched by age, race/ethnicity, and date of blood sampling. Serum CTX and serum PINP were analyzed on 12-hour fasting blood samples. The main outcome measure was incident hip fracture risk (mean follow-up 7.13 years). After adjustment for body mass index, smoking, frequency of falls, history of fracture, calcium and vitamin D intake, and other relevant covariates, neither serum CTX level nor serum PINP level was statistically significantly associated with hip fracture risk (CTX ptrend = 0.22, PINP ptrend = 0.53). Our results do not support the utility of serum CTX level or PINP level to predict hip fracture risk in women in this age group. These results will inform future guidelines regarding the potential utility of these markers in fracture prediction. © 2018 American Society for Bone and Mineral Research.


The use of continuous infusion topotecan in persistent and recurrent ovarian cancer.
We retrospectively review our experience with continuous infusion topotecan for the treatment of persistent or recurrent ovarian cancer in this paper. Nine patients were identified who were treated at the University of California Los Angeles Medical Center between January 1997 and December 1999 using a 14-21 day continuous infusion schedule (0.3-0.7 mg/m2/d). Dose adjustments were performed for grade 3-4 toxicities and treatment was discontinued for persistent severe toxicity or progressive disease. Response to treatment was analyzed and stratified by platinum refractory, resistant, and sensitive disease. A total of 41 treatment cycles were given to nine patients with a median of five per patient (range 1-11). Median follow-up was 8 months. There were two partial responses (22%) and four patients had stable disease (44%), which included two patients with platinum-refractory tumors. No grade 3 or 4 hematologic toxicities were observed. However, two patients suffered grade 3 gastrointestinal toxicity during the first cycle leading to discontinuation of topotecan administration. There was no cumulative toxicity. Topotecan administered by continuous infusion demonstrated response rates comparable to other dosing schedules with minimal hematologic toxicity. Treatment of patients with persistent or recurrent ovarian cancer with continuous infusion topotecan warrants further investigation.


Roseospira visakhapatnamensis sp. nov. and Roseospira goensis sp. nov.
Two Gram-negative, vibrioid, phototrophic, purple non-sulfur strains, JA131T and JA135T, were isolated from marine habitats. Strain JA131T is non-motile but strain JA135T is motile by means of a pair of monopolar flagella. Both strains have an obligate requirement for NaCl for growth. The intracellular photosynthetic membranes of the two novel strains are of the vesicular type. Bacteriochlorophyll a and probably rhodovibrine are present as photosynthetic pigments. Niacin, thiamine and p-aminobenzoic acid are required as growth factors for both novel strains. Based on 16S rRNA gene sequence analysis, morphological and physiological characteristics, strains JA131T and JA135T are significantly different from each other and from other species of the genus Roseospira and thus represent two novel species for which the names Roseospira visakhapatnamensis sp. nov. and Roseospira goensis sp. nov. are proposed, respectively. The type strain of Roseospira visakhapatnamensis sp. nov. is JA131T (=ATCC BAA-1365T=JCM 14190T) and the type strain of Roseospira goensis sp. nov. is JA135T (=ATCC BAA-1364T=JCM 14191T).


A new approach for the direct visualization of the membrane cytoskeleton in cryo-electron microscopy: a comparative study with freeze-etching electron microscopy.
An improved unroofing method consisting of tearing off the cell membrane using an adhesive electron microscopy (EM) grid instead of vitreous ice sectioning (cryo-sectioning) has enabled us to panoramically view the membrane cytoskeleton in its native state with extremely high contrast. Grids pre-treated with Alcian blue were placed on cells, and a portion of the dorsal plasma membrane was transferred onto the grid, which was then floated in buffer solution. These membrane fragments contained sufficient cytoskeleton and were of suitable thickness for observation by cryo-EM. Many actin filaments and microtubules were clearly observed on the cytoplasmic surface of the plasma membrane with extremely high contrast because the soluble components of the cytoplasm flowed out and broke away from the cells. Actin filaments extended in all directions in a smooth contour with little branching. Microtubules spread out as far as 3 µm or more while winding gently in their native state. Upon fixation with 1% glutaraldehyde, however, the microtubules became straight and fragmented. Cryo-EM revealed for the first time a smooth endoplasmic reticulum network beneath the cell membrane in native cells. Clathrin coats and caveolae were also observed on the cytoplasmic surface of the plasma membrane, similar to those seen using freeze-etching replica EM (freeze-etching EM). Unroofing was also useful for immuno-labelling in cryo-EM. Antibody-labelled IQGAP1, one of the effector proteins facilitating the formation of actin filament networks, was localized alongside actin filaments. Freeze-etching EM confirmed the morphological findings of cryo-EM.


Nse1, Nse2, and a novel subunit of the Smc5-Smc6 complex, Nse3, play a crucial role in meiosis.
The structural maintenance of chromosomes (SMC) family of proteins play key roles in the organization, packaging, and repair of chromosomes. Cohesin (Smc1+3) holds replicated sister chromatids together until mitosis, condensin (Smc2+4) acts in chromosome condensation, and Smc5+6 performs currently enigmatic roles in DNA repair and chromatin structure. The SMC heterodimers must associate with non-SMC subunits to perform their functions. Using both biochemical and genetic methods, we have isolated a novel subunit of the Smc5+6 complex, Nse3. Nse3 is an essential nuclear protein that is required for normal mitotic chromosome segregation and cellular resistance to a number of genotoxic agents. Epistasis with Rhp51 (Rad51) suggests that like Smc5+6, Nse3 functions in the homologous recombination based repair of DNA damage. We previously identified two non-SMC subunits of Smc5+6 called Nse1 and Nse2. Analysis of nse1-1, nse2-1, and nse3-1 mutants demonstrates that they are crucial for meiosis. The Nse1 mutant displays meiotic DNA segregation and homologous recombination defects. Spore viability is reduced by nse2-1 and nse3-1, without affecting interhomolog recombination. Finally, genetic interactions shared by the nse mutants suggest that the Smc5+6 complex is important for replication fork stability.


Parasternal and external intercostal muscle shortening during eupneic breathing.
The interosseous external intercostal (EI) muscles of the upper rib cage are electrically active during inspiration, but the mechanical consequence of their activation is unclear. In 16 anesthetized dogs, we simultaneously measured EI (3rd and 4th interspaces) and parasternal intercostal (PA) (3rd interspace) electromyogram and length. Muscle length was measured by sonomicrometry and expressed as a percentage of resting length (%LR). During resting breathing, each muscle was electrically active and shortened to a similar extent. Sequential EI muscle denervation (3rd and 4th interspaces) followed by PA denervation (3rd interspace) demonstrated significant reductions in the degree of inspiratory shortening for each muscle. Mean EI muscle shortening of the third and fourth interspaces decreased from -3.4 +/- 0.5 and -3.0 +/- 0.4% LR (SE) under control conditions to -0.2 +/- 0.2 and -0.8 +/- 0.3% LR, respectively, after selective denervation of each of these muscles (P less than 0.001 for each). After selective denervation of the PA muscle, its shortening decreased from -3.5 +/- 0.3 to +0.6% LR (SE) (P less than 0.001). PA muscle denervation also caused the EI muscle in the third interspace to change from inspiratory shortening of -0.2% to inspiratory lengthening of +0.2% +/- 0.2 (P less than 0.05). We conclude that during eupneic breathing 1) the EI muscles of the upper rib cage, like the PA muscles, are inspiratory agonists and actively contribute to rib cage expansion and 2) PA muscle contraction contributes to EI muscle shortening.


Psychological and dietetic counselling combined in the treatment of obesity: a comparative study in a hospital outpatient clinic.
This study examined the effectiveness of combining behaviour modification with dietetic counselling in a hospital outpatient clinic. Thirty-six females with an obesity index over 25 were placed in one of three 16-week treatment conditions: (1) individual dietetic counselling, (2) group dietetic counselling, (3) group dietetic counselling with behaviour modification. Median end of treatment weight losses were 8.3, 4.6 and 6.9 kg respectively. At one year follow-up patients in Gp 2 tended to have regained their weight while patients in Gps 1 and 3 were equally successful in maintaining weight loss. However, trends towards lower rates of attrition, slower rates of decay of treatment effects and a more economical use of professional time favoured the dietetic behavioural group condition as a treatment choice. In view of the successful use of behavioural techniques by a variety of professions their wider employment by dietitians is advocated.


Female-biased obligate strategies in a partially migratory population.
Partial migration occurs when a breeding population consists of seasonal migrants and year-round residents. Although it is common among birds, the basis of individual movement decisions within partially migratory populations is still unresolved. Over 4 years, we used state of the art tracking techniques, a combination of geolocators and radio transmitters, to follow individual European blackbirds Turdus merula year round from a partially migratory population to determine individual strategies and departure and arrival dates. The individual-based tracking combined with measures of energetic and hormonal (corticosterone) state enabled us to distinguish between obligate and facultative migration and to test several classical hypotheses of partial migration: the 'Arrival Time'-, 'Dominance'- and 'Thermal Tolerance'-hypotheses. Two distinct periods of departures from the breeding grounds were observed during the study; one in early autumn, and another during the midst of winter. Although blackbirds that migrated in autumn were never observed overwintering within 300 km of the study site, four individuals that departed in the winter were observed within 40 km. Females were significantly more likely to migrate in autumn than males but there was no difference in the age or body size of migrants and non migrants in autumn. Just prior to autumn migration, migrants had higher fat scores than non migrants and tended to have higher concentrations of baseline corticosterone, but similar concentrations of triglycerides. Unlike autumn migrants, we found no difference between the tendencies of males versus females to depart in winter, nor did we find any difference in body size or age of individuals that departed in the winter. Autumn migration was sex biased and resembled obligate migration. Our results provide strong support for the 'Arrival Time' hypothesis for partial migration in the autumn. We found no clear support for the 'Dominance' or 'Thermal Tolerance' hypotheses. By tracking individuals year round, we were able to identify a second period of departures. Overall, these results suggest the co-occurrence of obligate autumn migrants, winter movements and sedentary individuals within a single population.


Multiple-subunit genes of the aromatic-ring-hydroxylating dioxygenase play an active role in biphenyl and polychlorinated biphenyl degradation in Rhodococcus sp. strain RHA1.
A gram-positive strong polychlorinated biphenyl (PCB) degrader, Rhodococcus sp. strain RHA1, can degrade PCBs by cometabolism with biphenyl or ethylbenzene. In RHA1, three sets of aromatic-ring-hydroxylating dioxygenase genes are induced by biphenyl. The large and small subunits of their terminal dioxygenase components are encoded by bphA1 and bphA2, etbA1 and etbA2, and ebdA1 and ebdA2, respectively, and the deduced amino acid sequences of etbA1 and etbA2 are identical to those of ebdA1 and ebdA2, respectively. In this study, we examined the involvement of the respective subunit genes in biphenyl/PCB degradation by RHA1. Reverse transcription-PCR and two-dimensional polyacrylamide gel electrophoresis analyses indicated the induction of RNA and protein products of etbA1 and ebdA1 by biphenyl. Single- and double-disruption mutants of etbA1, ebdA1, and bphA1 were constructed by insertional inactivation. The 4-chlorobiphenyl (4-CB) degradation activities of all the mutants were lower than that of RHA1. The results indicated that all of these genes are involved in biphenyl/PCB degradation. Furthermore, we constructed disruption mutants of ebdA3 and bphA3, encoding ferredoxin, and etbA4, encoding ferredoxin reductase components. The 4-CB degradation activities of these mutants were also lower than that of RHA1, suggesting that all of these genes play a role in biphenyl/PCB degradation. The substrate preferences of etbA1A2/ebdA1A2- and bphA1A2-encoded dioxygenases for PCB congeners were examined using the corresponding mutants. The results indicated that these dioxygenase isozymes have different substrate preferences and that the etbA1A2/ebdA1A2-encoded isozyme is more active on highly chlorinated congeners than the bphA1A2-encoded one.


Mycobacterium avium infection in a silicone-injected breast.
A case of atypical mycobacterial infection (M. avium intracellulare) in a silicone-injection augmented breast is described. The silicone injection may have been a contributing factor to the development of this unusual infection. Disseminated M. avium was present in this patient, with breast involvement being suggested by the rapid appearance and disappearance of localized areas of erythema and tenderness. Aggressive treatment of these breast infections while they are still localized may prevent systemic spread. Conventional incision and drainage of the breast abscess combined with multidrug treatment directed against M. avium is the recommended therapy.


Structurally different neuronal nicotinic acetylcholine receptor subtypes purified and characterized using monoclonal antibodies.
Acetylcholine receptors that bind nicotine with high affinity but do not bind alpha-bungarotoxin have recently been immunoaffinity purified from brains of chickens and rats (Whiting and Lindstrom, 1986a, b; Whiting and Lindstrom, 1987a). Antisera to these receptors bind to the nicotinic receptors that regulate cation channel opening on chick ciliary ganglion neurons (Stollberg et al., 1986) and rat PC12 cells (Whiting et al., 1987c). Here we report the preparation and characterization of monoclonal antibodies to chicken brain acetylcholine receptors. These monoclonal antibodies are used to identify 2 nicotinic receptor subtypes in the chicken brain. The 2 subtypes have very similar affinities for nicotine and other cholinergic agonists and antagonists. However, they are structurally distinct, having very similar or identical alpha subunits (Mr 49,000), but different beta subunits (Mr 59,000, or for beta' subunit, Mr 75,000). Evidence is presented that suggests that the subunit stoichiometry of these neuronal nicotinic acetylcholine receptors is alpha n = 2 - 3 beta n = 2 - 3. Different levels of receptor subtype expression were detected in embryonic, compared to adult, chicken brain.


[3H]Pirenzepine and [3H]quinuclidinyl benzilate binding to brain muscarinic cholinergic receptors. Differences in measured receptor density are not explained by differences in receptor isomerization.
Muscarinic receptor densities were measured in membranes prepared from rat cerebral cortex using [3H]pirenzepine and [3H]quinuclidinyl benzilate. Isotherms of equilibrium binding data modeled to a single apparent binding site for both ligands. However, as has been reported previously, [3H]pirenzepine labeled only a small fraction of the binding sites that were labeled by [3H]quinuclidinyl benzilate. This observation has been used to support the hypothesis that subtypes of muscarinic receptors exist. Several investigators have previously suggested that antagonist binding to muscarinic receptors involves an isomerization of the receptor-antagonist complex, and it is only the isomerized form of the receptor that is identified by radioligand binding studies. To examine the possibility that the difference in the density of binding sites identified by [3H]pirenzepine and [3H]quinuclidinyl benzilate is due to differences in the degree of isomerization of the receptor associated with the binding of each ligand, the kinetics of the binding of [3H]pirenzepine and [3H]quinuclidinyl benzilate to membranes prepared from rat cerebral cortex were examined. The pseudo-first-order rate constant of association for both ligands showed a nonlinear (hyperbolic) dependence on ligand concentration. These results suggested that a rapidly equilibrating initial binding step was followed by a more slowly equilibrating isomerization of the initially formed ligand-receptor complex. The kinetic data were computer-modeled to obtain estimates of the equilibrium constants for both reaction steps. The equilibrium constants for the isomerization step were 0.1 and 0.004 for [3H]pirenzepine and [3H]quinuclidinyl benzilate, respectively. Our measurements, in agreement with others, suggested that only the fraction of receptors which isomerized were measurable using filtration binding assays. Although essentially all (99.6%) of the [3H]quinuclidinyl benzilate binding sites appeared to isomerize, only 90% of the [3H]pirenzepine binding sites isomerized, and thus only 90% were measured in our assay. It therefore appears that differences in receptor isomerization can partially, but not wholly, account for the differences between [3H]pirenzepine and [3H]quinuclidinyl benzilate binding in rat cerebral cortex.


Activation and inhibition of the Escherichia coli F1-ATPase by monoclonal antibodies which recognize the epsilon subunit.
The properties of two monoclonal antibodies which recognize the epsilon subunit of Escherichia coli F1-ATPase were studied in detail. The epsilon subunit is a tightly bound but dissociable inhibitor of the ATPase activity of soluble F1-ATPase. Antibody epsilon-1 binds free epsilon with a dissociation constant of 2.4 nM but cannot bind epsilon when it is associated with F1-ATPase. Likewise epsilon cannot associate with F1-ATPase in the presence of high concentrations of epsilon-1. Thus epsilon-1 activates F1-ATPase which contains the epsilon subunit, and prevents added epsilon from inhibiting the enzyme. Epsilon-1 cannot bind to membrane-bound F1-ATPase. The epsilon-4 antibody binds free epsilon with a dissociation constant of 26 nM. Epsilon-4 can bind to the F1-ATPase complex, but, like epsilon-1, it reverses the inhibition of F1-ATPase by the epsilon subunit. The epsilon subunit remains crosslinkable to both the beta and gamma subunits in the presence of epsilon-4, indicating that it is not grossly displaced from its normal position by the antibody. Presumably the activation arises from more subtle conformational effects. Antibodies epsilon-4 and delta-2, which recognizes the delta subunit, both bind to F1F0 in E. coli membrane vesicles, indicating that these subunits are substantially exposed in the membrane-bound complex. Epsilon-4 inhibits the ATPase activity of the membrane-bound enzyme by about 50%, and Fab prepared from epsilon-4 inhibits by about 40%. This inhibition is not associated with any substantial change in the major apparent Km for ATP. These results suggest that inhibition of membrane-bound F1-ATPase arises from steric effects of the antibody.