Adaptation of lettuce mosaic virus to Catharanthus roseus involves mutations in the central domain of the VPg.
An isolate of Lettuce mosaic virus (LMV, a Potyvirus) infecting Madagascar periwinckle (Catharanthus roseus) was identified and characterized by Illumina deep sequencing. LMV-Cr has no close affinities to previously sequenced LMV isolates and represents a novel, divergent LMV clade. Inoculation experiments with other representative LMV isolates showed that they are unable to infect C. roseus, which was not known to be a host for LMV. However, three C. roseus variants of one of these isolates, LMV-AF199, could be selected and partially or completely sequenced. These variants are characterized by the accumulation of mutations affecting the C-terminal part of the cylindrical inclusion (CI) helicase and the central part of the VPg. In particular, a serine to proline mutation at amino acid 143 of the VPg was observed in all three independently selected variants and is also present in the LMV-Cr isolate, making it a prime candidate as a host-range determinant. Other mutations at VPg positions 65 and 144 could also contribute to the ability to infect C. roseus. Inoculation experiments involving a recombinant LMV expressing a permissive lettuce eukaryotic translation initiation factor 4E (eIF4E) suggest that eIF4E does not contribute to the interaction of most LMV isolates with C. roseus.


Lonely But Not Alone: Neuroticism Mediates the Relationship Between Social Network Size and Loneliness in Individuals With Traumatic Brain Injury.
Objectives: Although individuals with traumatic brain injury (TBI) often report higher levels of social isolation, little is known about the factors influencing their self-perception of loneliness. The aim of the current study was to investigate the relationship between loneliness, social network size, and personality variables (neuroticism and extraversion) after TBI, and in particular whether specific personality variables mediate the relationship between social network size and perception of loneliness. Methods: Here, we assessed self-reported loneliness, personality variables, and social network size of 24 individuals with moderate-to-severe TBI and 41 healthy comparison participants. We then carried out a mediation analysis to examine whether personality variables mediated the relationship between loneliness and social network size. Results: Our results indicate that individuals with TBI reported higher levels of loneliness and neuroticism, but there was no group difference in social network size or extraversion. The mediation analysis revealed that the association between social network size and loneliness was mediated by neuroticism, but not by extraversion. Conclusions: Our findings show that neuroticism is an intervening variable in the relationship between social network size and self-perception of loneliness in individuals with moderate-to-severe TBI, and presents a new possible target for clinicians and rehabilitators seeking to address reports of loneliness and social isolation in TBI. (JINS 2019, 25, 266-274).


A systematically structured review of biomarkers of dying in cancer patients in the last months of life; An exploration of the biology of dying.
The Neuberger review made a number of recommendations to improve end of life care, including research into the biology of dying. An important aspect of the biology of dying is the identification of biomarkers as indices of disease processes. Biomarkers have the potential to inform the current, limited understanding of the dying process and assist clinicians in recognising dying, in particular how to distinguish dying from reversible acute deterioration. To critically appraise the literature on biological factors that may be used as prognostic indicators in advanced cancer patients and to identify candidate biomarkers of the dying process that can be measured serially in cancer patients' bodily fluids. A systematically structured review was conducted using three electronic databases. A hand search of six peer-reviewed journals and conference abstracts was also conducted. Studies reporting prognostic biomarkers in cancer patients with a median survival of ≤90 days and post-mortem studies were included. Final levels of evidence and recommendations were made using the Evidence Based Medicine modified GRADE system. 30 articles were included. Seven prognostic biological factors demonstrated Grade A evidence (lymphocyte count, white blood cell count, serum C-reactive protein, albumin, sodium, urea and alkaline phosphatase). An additional eleven prognostic factors were identified with Grade B evidence (platelet count, international normalised ratio, serum vitamin B12, prealbumin, bilirubin, cholesterol, aspartate aminotransferase, alanine transaminase, lactate dehydrogenase, pseudocholinesterase and urate). A number of biomarkers were specifically identified in the last two weeks of life but limitations exist. No post-mortem studies met the inclusion criteria. The biology of dying is an important area for future research, with the evidence focused on signs, symptoms and prognostic factors. This review identifies a number of common themes shared amongst advanced cancer patients and highlights candidate biomarkers which may be indicative of a common biological process to dying.


[The divergence of the axonal collaterals of the thalamic neurons to the visual and associative cortexes in cats].
Projections of the thalamic neurons to the visual (area 17) and the parietal association (area 7) cortices were examined by retrograde axonal transport of fluorescent dyes. It was found that the pulvinar neurons can be divided into three groups with respect to their connections with these cortical areas: 1--projecting to area 7 (the largest cell group); 2--projecting to area 17 (a smaller cell group) and 3--sending their axons to the both cortical areas (only few cells). Neurons belonging only to first two groups were found in the lateral posterior nucleus. Divergence of axonal collaterals of pulvinar neurons can secure the existence of parallel pathways transmitting information to the visual and associative cortices.


Submucous cleft palate: from diagnosis to management
The aim of this study was to investigate submucous cleft palate (SMCP) patients in order to document the age of diagnosis, cause of referral, symptoms and palate findings, objective evaluation of nasalance and velopharyngeal dysfunction (VPD), and intervention type. The archive of Hacettepe University Cleft Lip and Palate, Craniomaxillofacial Anomalies Research, Treatment, and Application Center was reviewed retrospectively, and 166 patients diagnosed with SMCP were documented. The mean age of the patients at the time of initial referral was 10 years and 3 months ± 8 years and 5 months with the youngest being 1 month and the oldest 44 years old. The primary complaint of 127 (76.5%) patients was speech disorder. Seventy-nine patients underwent surgery, and the mean age was determined as 10 years ± 6 years and 8 months. The diagnosis of SMCP continues to be rather late, being delayed until a prominent speech disorder is present. Studies for the awareness of health professionals are of great importance for the early diagnosis and intervention of SMCP in order to prevent adverse effects.


A Single Episode of Hypoglycemia as a Possible Early Warning Sign of Adrenal Insufficiency.
A 65-year-old woman without a history of diabetes mellitus was admitted for elective total knee arthroplasty for osteoarthrosis. There were no specific complaints except for knee flexion contractures, and the results of preoperative tests were unremarkable. On the day of surgery, the patient suffered from a hypoglycemic attack (52 mg/dL) after preoperative overnight fasting. A dextrose infusion immediately corrected the hypoglycemia, and a total knee arthroplasty was then performed. Although a hypoglycemic attack did not recur, further evaluation was required because of nausea that persisted after surgery. The morning serum cortisol level was 0.15 µg/dL with undetectable adrenocorticotropic hormone (ACTH), and the insulin-like growth factor-1 level was 9 ng/mL. An empty sella and bilateral adrenal atrophy were evident in imaging studies. ACTH and growth hormone (GH) did not respond to testing with corticotropin-releasing hormone and GH-releasing peptide-2, respectively. While serum cortisol did not increase on a rapid ACTH stimulation test, urinary free cortisol excretion responded to a prolonged ACTH stimulation test. Finally, the patient was diagnosed as having empty sella syndrome with ACTH and GH deficiencies. After the administration of hydrocortisone as maintenance replacement therapy, the patient's prolonged postoperative nausea disappeared. Adrenal insufficiency is latent in patients with hypoglycemia episodes. Because patients with adrenal insufficiency require appropriate perioperative corticosteroid supplementation, clinicians should give priority to identifying the underlying etiology of hypoglycemia over non-urgent elective surgery when these co-occur.


Synthesis, structural and magnetic characterisation of iron(II/III), cobalt(II) and copper(II) cluster complexes of the polytopic ligand: N-(2-pyridyl)-3-carboxypropanamide.
Herein we describe the synthesis, structural and magnetic characterisation of three transition metal cluster complexes that feature the polytopic ligand N-(2-pyridyl)-3-carboxypropanamide (H2L): [Fe3(III)Fe2(II)(HL)6(O)(H2O)3][ClO4]5·3MeCN·4H2O, 1, [Co8(HL)8(O)(OH)4(MeOH)3(H2O)]-[ClO4]3·5MeOH·2H2O, 2, and [Cu6(L(ox))4(MeOH)(H2O)3]·MeOH, 3. Complex 1 is a mixed valence penta-nuclear iron cluster containing the archetypal {Fe3(III)O} triangular basic carboxylate cluster at its core, with two Fe(II) ions above and below the core coordinated to three bidentate pyridyl-amide groups. The structure of the octanuclear Co(II) complex, 2, is based upon a central Co4 square with the remaining four Co(II) centres at the 'wing-tips' of the complex. The cluster core is replete with bridging oxide, hydroxide and carboxylate groups. Cluster 3 contains an oxidised derivative of the ligand, L(ox), generated in situ through hydroxylation of an α-carbon atom. This hexanuclear cluster has a 'barrel-like' core and contains Cu(II) ions in both square planar and square-based pyramidal geometries. Bridging between Cu(II) centres is furnished by alkoxide and carboxylate groups. Magnetic studies on 1-3 reveals dominant antiferro-magnetic interactions for 1 and 2, leading to small non-zero spin ground states, while 3 shows ferro-magnetic exchange between the Cu(II) centres to give an S = 3 spin ground state.


Immobilization of enzymes on porous silicas--benefits and challenges.
Porous silica materials have extensively been used for the immobilization of enzymes aiming at their use as biocatalysts or biosensors. This tutorial review will discuss the benefits and challenges of different immobilization techniques and will provide references to recent papers for further reading. Moreover, novel trends and unsolved problems will be introduced.


Ability of self-reported estimates of dietary sodium, potassium and protein to detect an association with general and abdominal obesity: comparison with the estimates derived from 24 h urinary excretion.
As under-reporting of dietary intake, particularly by overweight and obese subjects, is common in dietary surveys, biases inherent in the use of self-reported dietary information may distort true diet-obesity relationships or even create spurious ones. However, empirical evidence of this possibility is limited. The present cross-sectional study compared the relationships of 24 h urine-derived and self-reported intakes of Na, K and protein with obesity. A total of 1043 Japanese women aged 18-22 years completed a 24 h urine collection and a self-administered diet history questionnaire. After adjustment for potential confounders, 24 h urine-derived Na intake was associated with a higher risk of general obesity (BMI≥25 kg/m2) and abdominal obesity (waist circumference≥80 cm; both P for trend=0·04). For 24 h urine-derived protein intake, positive associations with general and abdominal obesity were observed (P for trend=0·02 and 0·053, respectively). For 24 h urine-derived K intake, there was an inverse association with abdominal obesity (P for trend=0·01). Conversely, when self-reported dietary information was used, only inverse associations between K intake and general and abdominal obesity were observed (P for trend=0·04 and 0·02, respectively), with no associations of Na or protein intake. In conclusion, we found positive associations of Na and protein intakes and inverse associations of K intake with obesity when using 24 h urinary excretion for estimating dietary intakes. However, no association was observed based on using self-reported dietary intakes, except for inverse association of K intake, suggesting that the ability of self-reported dietary information using the diet history questionnaire for investigating diet-obesity relationships is limited.


Effective condylar growth and chin position changes in activator treatment: a cephalometric roentgenographic study.
Effective condylar growth (a summation of condylar remodeling, glenoid fossa remodeling, and condylar position changes within the fossa) and its influence on the position of the chin was analyzed in 40 class II, division I malocclusions treated successfully with activators. Additionally, the amount of mandibular rotation was assessed. Lateral head films in habitual occlusion from before and after an average treatment period of 2.6 years were evaluated. The Bolton Standards (32 untreated individuals with ideal occlusion) served as controls. Two different treatment effects were evaluated: overall growth change and treatment effects (overall growth changes minus age-related Bolton values). In comparison with the Bolton group, the activator patients exhibited an increase in the amount of vertical effective condylar growth (3.0 mm; P < .001), a decrease in the amount of sagittal effective condylar growth (0.6 mm; P < .05), and an increase in the amount of vertical development of the chin (1.8 mm; P < .001). No group differences could be found for sagittal development of the chin. In the Bolton group, the mandible rotated posteriorly, and in the activator group it rotated anteriorly (2.7 degrees; P < .001). The present investigation revealed that effective condylar growth can be increased and the chin position can be changed by activator treatment. Thus activator treatment induces skeletal changes, although not always in the desired (sagittal) therapeutic direction.


Regulation of glycolytic rate in Streptococcus sanguis grown under glucose-limited and glucose-excess conditions in a chemostat.
The biochemical mechanisms of the acidogenic potential of Streptococcus sanguis ATCC 10556 grown in glucose-excess and glucose-limited continuous culture were studied. The rate of acid production during the glucose metabolism by the cells grown under glucose limitation (glucose-limited cells) was 2.1 to 2.6 times that by the cells grown in an excess of glucose (glucose-excess cells). When the glucose-limited cells were metabolizing glucose, intracellular concentrations of glucose 6-phosphate, fructose 6-phosphate, 3-phosphoglycerate, and pyruvate were higher, and that of glyceraldehyde 3-phosphate was lower, than those when the glucose-excess cells were metabolizing glucose. The levels of fructose 1,6-bisphosphate and dihydroxyacetone phosphate were not significantly different between these cells. The activities of glucose-phosphoenolpyruvate phosphotransferase system in decriptified cells and glyceraldehyde-3-phosphate dehydrogenase in cell-free extracts of the glucose-limited cells were higher than those in the glucose-excess cells. The activities of glucokinase, phosphoglycerate kinase, and pyruvate kinase in cell-free extracts of these cells were not different significantly. We conclude that the high glycolytic activity of the glucose-limited cells results from the increase in the synthesis of glucose-phosphoenolpyruvate phosphotransferase and glyceraldehyde-3-phosphate dehydrogenase.


Computational Screening of Neuropilin-1 Unveils Novel Potential Anti-SARS-CoV-2 Therapeutics.
Neuropilin 1 (NRP-1) inhibition has shown promise in reducing the infectivity of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) and preventing the virus entry into nerve tissues, thereby mitigating neurological symptoms in COVID-19 patients. In this study, we employed virtual screening, including molecular docking, Molecular Dynamics (MD) simulation, and Molecular Mechanics-Poisson Boltzmann Surface Area (MM-PBSA) calculations, to identify potential NRP-1 inhibitors. From a compendium of 1930 drug-like natural compounds, we identified five potential leads: CNP0435132, CNP0435311, CNP0424372, CNP0429647, and CNP0427474, displaying robust binding energies of -8.2, -8.1, -10.7, -8.2, and -8.2 kcal/mol, respectively. These compounds demonstrated interactions with critical residues Tyr297, Trp301, Thr316, Asp320, Ser346, Thr349, and Tyr353 located within the b1 subdomain of NRP-1. Furthermore, MD simulations and MM-PBSA calculations affirmed the stability of the complexes formed, with average root mean square deviation, radius of gyration, and solvent accessible surface area values of 0.118 nm, 1.516 nm, and 88.667 nm2 , respectively. Notably, these lead compounds were estimated to penetrate the blood-brain barrier and displayed antiviral properties, with Pa values ranging from 0.414 to 0.779. The antagonistic effects of these lead compounds merit further investigation, as they hold the potential to serve as foundational scaffolds for the development of innovative therapeutics aimed at reducing the neuroinfectivity of SARS-CoV-2.


[Fluorometholon/tetryzoline in the treatment of conjunctival irritation].
The clinical efficacy of Efemoline eyedrops [fluourometholone (0.1%) + tetryzoline (0.25%)] and of fluorometholone 0.1% was investigated in a double-blind controlled clinical trial in 35 patients (63 eyes) in treatment of the upper bulbus area and postoperatively after surgery for cataract, glaucoma and strabismus. Both drugs were effective inflammation inhibitors. The superiority of fluorometholone + tetryzoline over fluorometholone was evident: subjective symptoms improved much more quickly. No tendency toward elevation of intraocular pressure was found. The combined preparation was compatible and produced no side-effects.


Strain-induced alignment in collagen gels.
Collagen is the most abundant extracellular-network-forming protein in animal biology and is important in both natural and artificial tissues, where it serves as a material of great mechanical versatility. This versatility arises from its almost unique ability to remodel under applied loads into anisotropic and inhomogeneous structures. To explore the origins of this property, we develop a set of analysis tools and a novel experimental setup that probes the mechanical response of fibrous networks in a geometry that mimics a typical deformation profile imposed by cells in vivo. We observe strong fiber alignment and densification as a function of applied strain for both uncrosslinked and crosslinked collagenous networks. This alignment is found to be irreversibly imprinted in uncrosslinked collagen networks, suggesting a simple mechanism for tissue organization at the microscale. However, crosslinked networks display similar fiber alignment and the same geometrical properties as uncrosslinked gels, but with full reversibility. Plasticity is therefore not required to align fibers. On the contrary, our data show that this effect is part of the fundamental non-linear properties of fibrous biological networks.


Neural responses associated with attentional engagement and disengagement from threat in high socially anxious children: Evidence from temporal-spatial PCA.
Cognitive models indicated that social anxiety disorder (SAD) would be caused and maintained by a biased attentional processing of threatening information. This study investigates whether socially anxious children may present impaired attentional engagement and disengagement from negative emotional faces, as well as their underlying event-related potential responses. Fifteen children with high levels of social anxiety (HSA; 9 boys; mean age = 9.99y; SD = 1.14) and twenty low socially anxious children (LSA; 16 boys; mean age = 10.47y; SD = 1.17) participated in a spatial cueing task in which they had to detect targets following neutral/disgusted faces in a valid or invalid location. No group effect was reported on reaction times [p>.05]. However, electrophysiological data showed lower P3a amplitude in HSA children compared with the LSA group when processing facial stimuli. They also reported larger N2 amplitudes for valid-disgusted targets and a larger P3a amplitude for the invalid-disgusted ones. In terms of electrophysiological data, our results validated, the hypothesis of attentional disengagement difficulties in SAD children. We also confirm the idea that high levels of social anxiety are associated with cognitive control impairments and have a greater impact on the processing efficiency than on the performance effectiveness.


The use of animal models to investigate the pathogenesis of neuroinflammatory disorders of the central nervous system.
A major hurdle in the research of neuroinflammatory disorders of the central nervous system is the inaccessibility of the organ. Thus investigation is limited to end-stage disease and systemic changes that occur during disease progression, neither of which may reflect the pathological process in the central nervous system. These factors may explain the lack of effective therapies in multiple sclerosis, a common chronic inflammatory disease of the central nervous system. To overcome such limitations approaches using more relevant animal models have been developed to study pathological mechanisms as well as the design of rational therapeutic strategies. This review describes the animal models used to study pathological processes leading to inflammation within the central nervous system that may be operating in multiple sclerosis and the use of these models in the design of more rational therapeutic strategies. The clinical heterogeneity of multiple sclerosis as well as the finding of different pathological patterns suggests that multiple sclerosis may be a spectrum of diseases that may represent different pathological processes. This and the renewed interest in the extent of axonal damage has led to the development of more relevant animal models, such as those in nonhuman primates, that both reflect the spectrum of multiple sclerosis and allow the development of species-specific therapeutic approaches. While many animal models are available, the use of relevant animal models that mimic either the different forms of multiple sclerosis or the spectrum of multiple sclerosis is critical to examine those factors, for example genes or proteins, that are of pathogenic relevance and can be used as targets for therapy.


Enhancement of chemotherapeutic drug toxicity to human tumour cells in vitro by a subset of non-steroidal anti-inflammatory drugs (NSAIDs).
The effect on cytotoxicity of combining a range of clinically important non-steroidal anti-inflammatory drugs (NSAIDs) with a variety of chemotherapeutic drugs was examined in the human lung cancer cell lines DLKP, A549, COR L23P and COR L23R and in a human leukaemia line HL60/ADR. A specific group of NSAIDs (indomethacin, sulindac, tolmetin, acemetacin, zomepirac and mefenamic acid) all at non-toxic levels, significantly increased the cytotoxicity of the anthracyclines (doxorubicin, daunorubicin and epirubicin), as well as teniposide, VP-16 and vincristine, but not the other vinca alkaloids vinblastine and vinorelbine. A substantial number of other anticancer drugs, including methotrexate, 5-fluorouracil, cytarabine, hydroxyurea, chlorambucil, cyclophosphamide, cisplatin, carboplatin, mitoxantrone, actinomycin D, bleomycin, paclitaxel and camptothecin, were also tested, but displayed no synergy in combination with the NSAIDs. The synergistic effect was concentration dependent. The effect appears to be independent of the cyclo-oxygenase inhibitory ability of the NSAIDs, as (i) the synergistic combination could not be reversed by the addition of prostaglandins D2 or E2; (ii) sulindac sulphone, a metabolite of sulindac that does not inhibit the cyclooxygenase enzyme, was positive in the combination assay: and (iii) many NSAIDs known to be cyclo-oxygenase inhibitors, e.g. meclofenamic acid, diclofenac, naproxen, fenoprofen, phenylbutazone, flufenamic acid, flurbiprofen, ibuprofen and ketoprofen, were inactive in the combination assay. The enhancement of cytotoxicity was observed in a range of drug sensitive tumour cell lines, but did not occur in P-170-overexpressing multidrug resistant cell lines. However, in the HL60/ADR and COR L23R cell lines, in which multidrug resistance is due to overexpression of the multidrug resistance-associated protein MRP, a significant increase in cytotoxicity was observed in the presence of the active NSAIDs. Subsequent Western blot analysis of the drug sensitive parental cell lines, DLKP and A549, revealed that they also expressed MRP and reverse-transcription-polymerase chain reaction studies demonstrated that mRNA for MRP was present in both cell lines. It was found that the positive NSAIDs were among the more potent inhibitors of [3H]-LTC4 transport into inside-out plasma membrane vesicles prepared from MRP-expressing cells, of doxorubicin efflux from preloaded cells and of glutathione-S-transferase activity. The NSAIDs did not enhance cellular sensitivity to radiation. The combination of specific NSAIDs with anticancer drugs reported here may have potential clinical applications, especially in the circumvention of MRP-mediated multidrug resistance.


Abnormalities of enteric neurons, intestinal pacemaker cells, and smooth muscle in human intestinal atresia.
Intestinal dysmotility, which usually has been encountered in the severely dilated proximal segment, is an important problem in postoperative management of patients with intestinal atresia (IA). Changes of enteric nerves had been histochemically examined in both the proximal and distal segments of IA, but a systemic immunohistochemical analysis is still lacking. The aim of this study was to examine precisely alterations of neuronal and muscular elements and pacemaker cells in intestines from patients with IA. Resected intestines were obtained from 5 patients with ileal atresia, 3 patients with jejunal atresia, and 3 controls without gastrointestinal diseases (congenital diaphragmatic hernia). All specimens were immunochemically stained with a monoclonal antibody to alpha-smooth muscle actin (SMA) as a smooth muscle marker, polyclonal antibodies to protein gene product (PGP) 9.5 as a general neuronal marker, and to c-kit protein as a maker of intestinal pacemaker cells. In addition, all specimens also were stained by NADPH-diaphorase (NADPH-d) to know the distribution of inhibitory nitrergic nerves. A hypoplasia of the myenteric ganglia and a marked reduction of intramuscular nerve fibers, including nitrergic neurons, were observed in the dilated proximal segment of IA. C-kit-positive cells were localized around the myenteric plexus, but rarely found within the muscularis propria in the proximal segment. The distribution of nerves and c-kit-positive cells in the distal segment was comparable with that seen in controls. A reduced staining intensity for alpha-SMA was mainly observed in the hypertrophic circular muscle layer of the proximal segment. A hypoplasia of intramural nerves and pacemaker cells was seen predominantly in the proximal segments of IA. Hypertrophy and reduced immunoreactivity for alpha-SMA also were observed in the circular muscle layer of the proximal segment. These alterations of the proximal segment may thus contribute to the postoperative intestinal dysmotility in IA cases.


Uptake and Associated Factors of Male Contraceptive Method Use: A Community-Based Cross-Sectional Study in Northern Uganda.
Uganda has one of the highest fertility rates in Sub-Saharan Africa (SSA). Improving contraceptive uptake in all genders, including males, may be critical to meeting family planning goals in such a setting. Yet, data on male contraception uptake and associated factors in SSA, including Uganda, are limited. We determined the uptake and associated factors of male contraception use in Lira City, Northern Uganda. We conducted a community-based cross-sectional study from November 12, 2022, to December 12, 2022, among men aged ≥18 years. We used multi-stage sampling to select participants from 12 cells of Lira City divisions of East and West. Data were collected using interviewer-administered structured questionnaires. We defined uptake in males who had used any contraceptive method, including periodic abstinence, withdrawal, condoms, and vasectomy in the previous four weeks. We performed modified Poisson regression to identify associated factors of male contraception uptake. We recruited 401 participants with mean age of 30.4 (±9.3) years. Male contraceptive uptake was 46.4%, 95% CI: 41.5-51.3%. Ever heard about male contraception (adjusted prevalence ratio [aPR] =1.73, 95% CI: 1.172-2.539, p=0.006), willingness to use novel methods (aPR=2.90, 95% CI: 1.337-6.293, p=0.007), both partners being responsible for contraception (aPR: 1.53, 95% CI: 1.113-2.119, p=0.009) were the factors associated with male contraception uptake. We found that nearly half of the men surveyed had used male contraceptive methods in Lira City. Factors associated with the uptake of male contraception included having heard about male contraception, joint couple decision regarding contraception, and the use of novel methods of male contraception. We recommend comprehensive education and awareness campaigns to promote male contraception, with a particular emphasis on encouraging shared decision-making within couples and introducing innovative contraceptive options.


[Health problems among the Kaingáng (Xapecó Indigenous Reserve, Santa Catarina) and the health care system].
The second semester of 1999 was a transition period for the implementation of the Special Indigenous Health District on the Xapecó Indigenous Reserve in western Santa Catarina State. The health clinic in the main village provided treatment with a staff including a general practitioner/obstetrician, pediatrician, dentist, nurse, two nursing assistants, and four nursing technicians. This paper presents the preliminary results of research on the organization of these health care services, their use by the community, and the health/disease profile of the Kaingáng, using patient files as the source of information. In September 1999, a total of 222 Indians were treated (children and adults), 50.5% of whom residing in the main village. Among the Indians ages 0 to 14 years, infectious and parasitic diseases were the most frequent, supporting the idea that the Kaingáng have precarious sanitary and nutritional conditions. Use of the clinic by adults was more varied, since of the 116 who appeared for consultation, 27 were pregnant women (out of a total of 86 women). In addition, prescriptions were written up for children and adults in 85.0% and 81.8% of the consultations, respectively.


Two-flux radiative transfer model under nonisotropic propagating diffuse radiation.
A two-flux model is considered as a particular case of a more-general four-flux approach to describing the properties of highly diffusing materials derived from the radiative transfer equation. Any degree of anisotropy is taken into account by means of average path-length parameters and forward-scattering ratios for diffuse radiation propagating in forward and backward directions. The conditions for applicability of the standard two-flux model of Kubelka and Munk are characterized in terms of particle size and refractive index as well as of optical thickness. Scattering and absorption coefficients are obtained in terms of the effective average path-length parameter and forward-scattering ratio of the propagating radiation as well as of the intrinsic scattering and absorption coefficients per unit length of the particulate medium.


Liraglutide Treatment Ameliorates Neurotoxicity Induced by Stable Silencing of Pin1.
Post-translational modulation of peptidylprolyl isomerase Pin1 might link impaired glucose metabolism and neurodegeneration, being Pin1 effectors target for the glucagon-Like-Peptide1 analog liraglutide. We tested the hypotheses in Pin1 silenced cells (SH-SY5Y) treated with 2-deoxy-d-glucose (2DG) and methylglyoxal (MG), stressors causing altered glucose trafficking, glucotoxicity and protein glycation. Rescue by liraglutide was investigated. Pin1 silencing caused increased levels of reactive oxygen species, upregulated energy metabolism as suggested by raised levels of total ATP content and mRNA of SIRT1, PGC1α, NRF1; enhanced mitochondrial fission events as supported by raised protein expression of FIS1 and DRP1. 2DG and MG reduced significantly cell viability in all the cell lines. In Pin1 KD clones, 2DG exacerbated altered mitochondrial dynamics causing higher rate of fission events. Liraglutide influenced insulin signaling pathway (GSK3b/Akt); improved cell viability also in cells treated with 2DG; but it did not revert mitochondrial dysfunction in Pin1 KD model. In cells treated with MG, liraglutide enhanced cell viability, reduced ROS levels and cell death (AnnexinV/PI); and trended to reduce anti-apoptotic signals (BAX, BCL2, CASP3). Pin1 silencing mimics neuronal metabolic impairment of patients with impaired glucose metabolism and neurodegeneration. Liraglutide rescues to some extent cellular dysfunctions induced by Pin1 silencing.


A comprehensive phase I and II clinical investigation of fludarabine phosphate.
This article presents data from the phase I and II clinical investigations of Fludara I.V. (fludarabine phosphate) (NSC 312887), which is the 5'-phosphorylated derivative of the novel antimetabolite, 9-beta-D-arabinofuranosyl-2-fluoroadenine. The comprehensive phase I evaluation of this new antitumor agent was conducted in 51 patients with advanced malignancy and 15 additional patients with aggressive forms of leukemia. Three separate phase I schedules of drug administration were examined. Myelosuppression was the dose-limiting toxicity on each schedule administered to patients with solid tumors. The drug was also examined at higher doses in patients with leukemia, and the dose-limiting toxicity on the high-dose protocol was unacceptable: serious neurologic toxicity. The observation of antitumor responses in patients with advanced non-Hodgkin's lymphoma prompted additional phase II investigation in patients with lymphoproliferative malignancy. The encouraging phase II data demonstrate that Fludara I.V. has promise for patients with low-grade histologic subtypes of non-Hodgkin's lymphoma and chronic lymphocytic leukemia. While interesting additional basic and clinical research projects regarding Fludara I.V. remain, it is important to expeditiously pursue approval for this drug. Adequate data exists to demonstrate that the low-dose administration of Fludara I.V. is both safe and effective. While the development of this drug has stimulated renewed interest in the clinical investigation of the chronic lymphoproliferative malignancies, the time for making it readily available to these patients has arrived.


An investigation of the effect of electrode size and electrode location on comfort during stimulation of the gastrocnemius muscle.
The use of surface neuromuscular electrical stimulation (SNMES) in medicine is well established. However, discomfort has been identified as limiting the use of SNMES in these applications. This pilot study investigated the influence of various electrode sizes and their positioning on perceived pain and discomfort during neuromuscular electrical stimulation (NMES) of the gastrocnemius muscle using surface electrodes. This study formed part of a research project to develop a therapeutic device for calf muscle blood flow assist applications. Twelve healthy subjects (n=12) participated in this pilot study. Each participant attended the trial centre for testing which consisted of SNMES to four different electrode stimulation sites using two electrode sizes (round with areas 19.63 and 38.48 cm2). Comfort was assessed by asking the subjects to indicate the stimulation amplitude corresponding to the onset of discomfort (pain threshold) and the amplitude at which the discomfort became unbearable (pain tolerance). Of the four stimulation sites tested, two were deemed unsuccessful as it was very difficult to obtain a muscle contraction using these sites, while the remaining two sites elicited good muscle contraction. The most comfortable stimulation was achieved by placing the cathode electrode high on the calf, below the proximal end of the muscle heads and the anode electrode towards the end of the muscle belly and when the 19.63 cm2 electrodes were used at these sites (p=<0.001).


Landscape genetics of the nonnative red fox of California.
Invasive mammalian carnivores contribute disproportionately to declines in global biodiversity. In California, nonnative red foxes (Vulpes vulpes) have significantly impacted endangered ground-nesting birds and native canids. These foxes derive primarily from captive-reared animals associated with the fur-farming industry. Over the past five decades, the cumulative area occupied by nonnative red fox increased to cover much of central and southern California. We used a landscape-genetic approach involving mitochondrial DNA (mtDNA) sequences and 13 microsatellites of 402 nonnative red foxes removed in predator control programs to investigate source populations, contemporary connectivity, and metapopulation dynamics. Both markers indicated high population structuring consistent with origins from multiple introductions and low subsequent gene flow. Landscape-genetic modeling indicated that population connectivity was especially low among coastal sampling sites surrounded by mountainous wildlands but somewhat higher through topographically flat, urban and agricultural landscapes. The genetic composition of populations tended to be stable for multiple generations, indicating a degree of demographic resilience to predator removal programs. However, in two sites where intensive predator control reduced fox abundance, we observed increases in immigration, suggesting potential for recolonization to counter eradication attempts. These findings, along with continued genetic monitoring, can help guide localized management of foxes by identifying points of introductions and routes of spread and evaluating the relative importance of reproduction and immigration in maintaining populations. More generally, the study illustrates the utility of a landscape-genetic approach for understanding invasion dynamics and metapopulation structure of one of the world's most destructive invasive mammals, the red fox.


[Crystallography of 804 renal calculi].
We analyse a total of 804 kidney stones in 225 patients aged between 10 and 71 years, with an average age of around 45, by means of the following techniques: optico-differential analysis (immersion method, PRIEN and FRONDEL, 1947), fine plate, X-ray diffraction, infrared-ray spectroscopy and qualitative chemical analysis. In all the stones, we identify the composition of the nucleus and periphery separately. Our results give 51.32% of patients forming essentially oxalic stones, 28.31% uric ones, 19.46% phosphatic and 0.44% cystine stones. These statistics coincide quite closely with those obtained by Cifuentes (quot. in Dalet, F.; Pinto, B., 1976) in Madrid and differ a little more from those obtained by the Puigvert Foundation (quot. in Dalet, F.; Pinto, B., 1976) in Barcelona. They are also similar to those obtained by Sutor (1974) for non Anglo-Saxon, industrialized countries, such as Czechoslovakia and Kuwait. We identify the following components in this collection of stones: anhydrous uric acid, dihydrate uric acid, amorphous urates (sodic, ammonic), whewellite, wedellite, hydroxyapatite, carbo-apatite, struvite, newberyte, whitlockite, brushite and cystine. We found no xanthine stone.


A method for analysis and design of metabolism using metabolomics data and kinetic models: Application on lipidomics using a novel kinetic model of sphingolipid metabolism.
We present a model-based method, designated Inverse Metabolic Control Analysis (IMCA), which can be used in conjunction with classical Metabolic Control Analysis for the analysis and design of cellular metabolism. We demonstrate the capabilities of the method by first developing a comprehensively curated kinetic model of sphingolipid biosynthesis in the yeast Saccharomyces cerevisiae. Next we apply IMCA using the model and integrating lipidomics data. The combinatorial complexity of the synthesis of sphingolipid molecules, along with the operational complexity of the participating enzymes of the pathway, presents an excellent case study for testing the capabilities of the IMCA. The exceptional agreement of the predictions of the method with genome-wide data highlights the importance and value of a comprehensive and consistent engineering approach for the development of such methods and models. Based on the analysis, we identified the class of enzymes regulating the distribution of sphingolipids among species and hydroxylation states, with the D-phospholipase SPO14 being one of the most prominent. The method and the applications presented here can be used for a broader, model-based inverse metabolic engineering approach.


Rapid evolution of Medicago polymorpha during invasion shifts interactions with the soybean looper.
The Enemy Release Hypothesis posits that invasion of novel habitats can be facilitated by the absence of coevolved herbivores. However, a new environment and interactions with unfamiliar herbivores may impose selection on invading plants for traits that reduce their attractiveness to herbivores or for enhanced defenses compared to native host plants, leading to a pattern similar to enemy release but driven by evolutionary change rather than ecological differences. The Shifting Defense Hypothesis posits that plants in novel habitats will shift from specialized defense mechanisms to defense mechanisms effective against generalist herbivores in the new range. We tested these ideas by comparing herbivore preference and performance of native (Eurasia)- and invasive (New World)-range Medicago polymorpha, using a generalist herbivore, the soybean looper, that co-occurs with M. polymorpha in its New World invaded range. We found that soybean loopers varied in preference and performance depending on host genotype and that overall the herbivore preferred to consume plant genotypes from naïve populations from Eurasia. This potentially suggests that range expansion of M. polymorpha into the New World has led to rapid evolution of a variety of traits that have helped multiple populations become established, including those that may allow invasive populations to resist herbivory. Thus, enemy release in a novel range can occur through rapid evolution by the plant during invasion, as predicted by the Shifting Defense Hypothesis, rather than via historical divergence.


[A quantitative evaluation of blood-lymphocyte subpopulation in herpetic ocular disease].
Data on changing contents of blood-lymphocytes' subpopulations in patients with ophthalmoherpes are reported. A higher number of blood cells expressing the surface activation antigens of HLA-DR+, CD71+ and CD95+, a lower number of T-helpers and a reduced value of the immune-regulating index were found to be typical of such changes. A more significant increase in the quantity of CD95+ lymphocytes, as observed in cases of severe ophthalmoherpes versus its moderate clinical course, could be a prognostication criterion. The essential shifts occurring in the T-cells' population denote the status of tension in the immune response provoked by an intensified reproduction of herpes simplex virus (HSV); at the same time, they are indicative of a certain deficit in the immune-protection mechanisms.


Genetic Basis of Chronotype in Humans: Insights From Three Landmark GWAS.
Chronotype, or diurnal preference, refers to behavioral manifestations of the endogenous circadian system that governs preferred timing of sleep and wake. As variations in circadian timing and system perturbations are linked to disease development, the fundamental biology of chronotype has received attention for its role in the regulation and dysregulation of sleep and related illnesses. Family studies indicate that chronotype is a heritable trait, thus directing attention toward its genetic basis. Although discoveries from molecular studies of candidate genes have shed light onto its genetic architecture, the contribution of genetic variation to chronotype has remained unclear with few related variants identified. In the advent of large-scale genome-wide association studies (GWAS), scientists now have the ability to discover novel common genetic variants associated with complex phenotypes. Three recent large-scale GWASs of chronotype were conducted on subjects of European ancestry from the 23andMe cohort and the UK Biobank. This review discusses the findings of these landmark GWASs in the context of prior research. We systematically reviewed and compared methodological and analytical approaches and results across the three GWASs of chronotype. A good deal of consistency was observed across studies with 9 genes identified in 2 of the 3 GWASs. Several genes previously unknown to influence chronotype were identified. GWAS is an important tool in identifying common variants associated with the complex chronotype phenotype, the findings of which can supplement and guide molecular science. Future directions in model systems and discovery of rare variants are discussed.


Novel acid-catalyzed rearrangement of tetrahydro-1,2,3,4-tetrazines: unexpected formation of glycosazones.
The present contribution discloses a simple and unexpected acid-catalyzed cleavage of tetrahydrotetrazines leading to 1,2-bis(hydrazones). Incorporation of a chiral fragment derived from carbohydrates enables the rapid preparation of glycosazones, a family of compounds employed by Emil Fischer to elucidate the configuration of sugars. In addition, a mechanistic proposal accounts for experimental observations.


Comparison of the morphological and electrotonic properties of Renshaw cells, Ia inhibitory interneurons, and motoneurons in the cat.
The morphological and electrotonic properties of 4 motoneurons, 8 Ia inhibitory interneurons, and 4 Renshaw cells were compared. The morphological analysis, based on 3-D reconstructions of the cells, revealed that dendrites of motoneurons are longer and more extensively branched. Renshaw cells have dendrites that are shorter and simpler in structure. Dendrites of Ia inhibitory interneurons could be as long as those of motoneurons but the branching structure resembled that of Renshaw cells. Compartmental models were used to determine the electrotonic properties of the paths from each dendritic terminal to the soma. The attenuations of steady-state voltage changes in motoneurons were 3 and 7 times larger than in Ia inhibitory interneurons and Renshaw cells, respectively. The same relative order was observed for current attenuation and electrotonic length. The dendritic input resistances in Renshaw cells were 2 and 4 times larger than in Ia inhibitory interneurons and motoneurons, respectively. The difference in these electrotonic properties increased during higher synaptic activity as modeled by a decrease of Rm. The peak amplitudes of voltage transients at sites of brief, synaptic-like changes in conductance were highly dependent on cell class and were largest in Renshaw cells and smallest in motoneurons. In combination with class-specific differences in the attenuation of transient voltage signals, this led to large differences in the peak amplitudes of somatic voltage transients. Differences in the rise times and half-widths of the voltage transients were observed as well. Thus, based on passive properties, each cell class has a unique set of input/output properties.


Measurement of quality of life in cystic fibrosis.
Considerable progress has been made over the past 2 decades in defining and measuring health-related quality of life (QOL), and there is a growing recognition that these measures provide unique information about the impact of a chronic illness and its treatment. For patients with cystic fibrosis (CF), health-related QOL measures enable researchers and clinicians to determine the effects of clinical interventions on several aspects of daily living (psychological, emotional, social) that are not reflected in typical health indicators, eg, pulmonary functioning scores. Three types of health-related QOL measures have been developed: 1) utility measures, 2) health profiles, and 3) disease-specific measures. The purpose of each type of health-related QOL measure is described, and its application to patients with CF is reviewed. Although important descriptive information has been obtained from utility measures, eg, the Quality of Well-Being Scale, and health profiles, eg, the Nottingham Health Profile, both of these instruments have serious limitations. Disease-specific measures, similar to those developed for children and adults with asthma, seem to hold the greatest promise for advancing our understanding of the impact of CF on daily life and for evaluating the effectiveness of new clinical interventions. The Cystic Fibrosis Questionnaire is the only published disease-specific measure of health-related QOL for children, adolescents, and adults with CF. Its psychometric properties are briefly reviewed, and directions for future research are suggested.


8-Oxoguanine accumulation in aged female brain impairs neurogenesis in the dentate gyrus and major island of Calleja, causing sexually dimorphic phenotypes.
In mammals, including humans, MTH1 with 8-oxo-dGTPase and OGG1 with 8-oxoguanine DNA glycosylase minimize 8-oxoguanine accumulation in genomic DNA. We investigated age-related alterations in behavior, 8-oxoguanine levels, and neurogenesis in the brains of Mth1/Ogg1-double knockout (TO-DKO), Ogg1-knockout, and human MTH1-transgenic (hMTH1-Tg) mice. Spontaneous locomotor activity was significantly decreased in wild-type mice with age, and females consistently exhibited higher locomotor activity than males. This decrease was significantly suppressed in female but not male TO-DKO mice and markedly enhanced in female hMTH1-Tg mice. Long-term memory retrieval was impaired in middle-aged female TO-DKO mice. 8-Oxoguanine accumulation significantly increased in nuclear DNA, particularly in the dentate gyrus (DG), subventricular zone (SVZ) and major island of Calleja (ICjM) in middle-aged female TO-DKO mice. In middle-aged female TO-DKO mice, neurogenesis was severely impaired in SVZ and DG, accompanied by ICjM and DG atrophy. Conversely, expression of hMTH1 efficiently suppressed 8-oxoguanine accumulation in both SVZ and DG with hypertrophy of ICjM. These findings indicate that newborn neurons from SVZ maintain ICjM in the adult brain, and increased accumulation of 8-oxoguanine in nuclear DNA of neural progenitors in females is caused by 8-oxo-dGTP incorporation during proliferation, causing depletion of neural progenitors, altered behavior, and cognitive function changes with age.


Jejunal Diverticulosis Presented with Acute Abdomen and Diverticulitis Complication: A Case Report.
Jejunal diverticulosis is a rare, usually asymptomatic disease. Its incidence increases with age. If symptomatic, diverticulosis may cause life-threatening acute complications such as diverticulitis, perforation, intestinal hemorrhage and obstruction. In this report, we aimed to present a 67-year-old male patient with jejunal diverticulitis accompanying with abdominal pain and vomiting. A 67-year-old male patient complaining of epigastric pain for a week and nausea and fever for a day presented to our emergency department. Ultrasonographic examination in our clinic revealed diverticulum-like images with thickened walls adjacent to the small intestine loops, and increase in the echogenicity of the surrounding mesenteric fat tissue. Contrast-enhanced abdominal computed tomography showed multiple diverticula, thickened walls with showing contrast enhancement and adjacent jejunum in the left middle quadrant, increased density of the surrounding mesenteric fat tissue, and mesenteric lymph nodes. The patient was hospitalized by general surgery department with the diagnosis of jejunal diverticulitis. Conservative intravenous fluid administration and antibiotic therapy were initiated. Clinical symptoms regressed and the patient was discharged from hospital after 2 weeks. In cases of diverticulitis it should be kept in mind that in patients with advanced age and pain in the left quadrant of the abdomen, diverticular disease causing mortality and morbidity does not always originate from the colon but might also originate from the jejunum.


Score distributions of gapped multiple sequence alignments down to the low-probability tail.
Assessing the significance of alignment scores of optimally aligned DNA or amino acid sequences can be achieved via the knowledge of the score distribution of random sequences. But this requires obtaining the distribution in the biologically relevant high-scoring region, where the probabilities are exponentially small. For gapless local alignments of infinitely long sequences this distribution is known analytically to follow a Gumbel distribution. Distributions for gapped local alignments and global alignments of finite lengths can only be obtained numerically. To obtain result for the small-probability region, specific statistical mechanics-based rare-event algorithms can be applied. In previous studies, this was achieved for pairwise alignments. They showed that, contrary to results from previous simple sampling studies, strong deviations from the Gumbel distribution occur in case of finite sequence lengths. Here we extend the studies to multiple sequence alignments with gaps, which are much more relevant for practical applications in molecular biology. We study the distributions of scores over a large range of the support, reaching probabilities as small as 10^{-160}, for global and local (sum-of-pair scores) multiple alignments. We find that even after suitable rescaling, eliminating the sequence-length dependence, the distributions for multiple alignment differ from the pairwise alignment case. Furthermore, we also show that the previously discussed Gaussian correction to the Gumbel distribution needs to be refined, also for the case of pairwise alignments.


Polytetrafluoroethylene bars in stabilizing the reconstructed sternum for pectus excavatum operations in children.
Nine children with severe pectus excavatum deformity were operated on in the Department of Pediatric Surgery, Thoracic Unit, Ege University Faculty of Medicine, Izmir, Turkey. There were four boys and five girls, and the mean age at operation was 6.3 years. A modified Ravitch technique was performed by using two polytetrafluoroethylene bars placed retrosternally for fixation and stabilization of the reconstituted anterior chest wall. The bars were removed 6 months after the operation. Results were satisfactory in all cases, with no surgical complications.


Determinants of the Choice of Job Search Channels by the Unemployed Using a Multivariate Probit Model.
We investigate factors affecting the job search channels (newspapers, agency, informal network, direct contact, internet) of unemployed individuals in Italy using a multivariate probit model. We separately analyse unemployed individuals who lost their job or who are entering the labour market for the first time (i.e., with or without previous job experience, respectively). We use cross-sectional microdata covering the 2014-2018 period. We do not find important differences in the determinants in common between unemployed individuals with and without previous job experience (such as, age, education, and citizenship). The main difference between the two samples is in the composition, with more young people in the latter group (mainly individuals in the school-to-work transition) than in the first one. We find that better-educated unemployed people have more of a multichannel attitude when searching for jobs than their counterparts. Moreover, for unemployed individuals with previous job experience, the characteristics of their last job play a crucial role in influencing the choice of search channel(s). For example, unemployed individuals from high-skilled professions (that typically require the employment of better-educated individuals) choose the internet with a higher probability than other channels, while those from low-skilled professions and with experience in manufacturing have a higher probability of considering employment agencies than those in qualified professions in commercial activities and services. We have assumed that the unobservable factors affecting the decision process of a multichannel job search (estimated through the pairwise correlation coefficients across the residuals of the models) are represented by personality features of an individual, such as motivation, self-control, and preferences. A strong, positive, and significant correlation is found between job searching through the internet and direct contact with firms, but with some spatial differences between macro-areas of the country.


[Hepatic granulomas in a feverish patient].
Fever of unknown origin is a diagnostic challenge. When the diagnosis remains elusive despite extensive non-invasive diagnostic studies, it can be necessary to conduct more invasive procedures, such as liver biopsy. We report a male with fever of unknown origin who had tender enlarged liver and raised liver enzymes whose hepatic biopsy disclosed epithelioid granulomas, with the etiological diagnosis made by the positivity of Coxiella burnetii serology.


Pioglitazone, a PPAR-gamma ligand, provides protection from dextran sulfate sodium-induced colitis in mice in association with inhibition of the NF-kappaB-cytokine cascade.
Nuclear factor-kappaB-dependent up-regulation of inflammatory cytokines occurs in inflammatory bowel disease. We investigated the effect of pioglitazone, a peroxisome proliferator-activated receptor-gamma ligand, on dextran sulfate sodium-induced colonic mucosal injury and inflammation in mice. Acute colitis was induced in female mice receiving 0, 1, 3, and 10 mg/kg i.p. of pioglitazone daily. Colonic mucosal inflammation was evaluated chemically and histologically. Thiobarbituric acid-reactive substances and tissue-associated myeloperoxidase activity were measured in intestinal mucosa as indices of lipid peroxidation and neutrophil infiltration, respectively. Colonic mRNA expression of pro-inflammatory cytokines and inducible nitric oxide synthase was measured by reverse transcription-PCR and nuclear factor-kappaB activation was evaluated by electrophoretic mobility shift assay. Dextran sulfate sodium administration resulted in decreases in body weight and colon length and increases in lipid peroxide and neutrophil accumulation of the intestine. In contrast, co-administration with pioglitazone prevented these changes. Transcripts coding for pro-inflammatory cytokines and inducible nitric oxide were expressed in high levels after the development of colitis, and pioglitazone markedly reduced mRNA expression of these genes. DNA binding activity of nuclear factor-kappaB was markedly increased, whereas in pioglitazone co-treated intestines the effect was significantly reduced. These data suggest that peroxisome proliferator-activated receptor-gamma may be a novel therapeutic target for the therapy of inflammatory bowel disease.


Migration of mesonephric cells into the mammalian gonad depends on Sry.
In mammals, the primary step in male sex determination is the initiation of testis development which depends on the expression of the Y-linked testis determining gene, Sry. The mechanisms by which Sry controls this process are unknown. Studies showed that cell migration from the adjacent mesonephros only occurs into XY gonads; however, it was not known whether this effect depended on Sry, another Y-linked gene, or the presence of one versus two X chromosomes. Here we provide genetic proof that Sry is the only Y-linked gene necessary for cell migration into the gonad. Cell migration from the mesonephros into the differentiating gonad is consistently associated with Sty's presence and with testis cord formation, suggesting that cell migration plays a critical role in the initiation of testis cord development. The induction of cell migration represents the earliest signaling pathway yet assigned to Sry.


A nonpolymorphic class I gene in the murine major histocompatibility complex.
DNA sequence analysis of a class I gene (Q10), which maps to the Qa2,3 locus in the C57BL/10 (H-2b haplotype) mouse, reveals that it is almost identical to a cDNA clone (pH16) isolated from a SWR/J (H-2q haplotype) mouse liver cDNA library. Exon 5, in particular, has an unusual structure such that a polypeptide product is unlikely to be anchored in the cell membrane. Our findings suggest that the two sequences are derived from allelic class I genes, which are nonpolymorphic, in contrast to H-2K allelic sequences from the same mice, and they may encode liver-specific polypeptides of unknown function. Our previous studies indicate that the Q10 gene is a potential donor gene for the generation of mutations at the H-2K locus by inter-gene transfer of genetic information. Thus the lack of polymorphism in class I genes at the Q10 locus implies either that they are not recipients for such exchanges or that selective pressure prevents the accumulation of mutations in genes at this locus.


Potential Micronutrients and Phytochemicals against the Pathogenesis of Chronic Obstructive Pulmonary Disease and Lung Cancer.
Lung cancer and chronic obstructive pulmonary disease have shared etiology, including key etiological changes (e.g., DNA damage and epigenetics change) and lung function impairment. Focusing on those shared targets may help in the prevention of both. Certain micronutrients (vitamins and minerals) and phytochemicals (carotenoids and phenols) have potent antioxidant or methyl-donating properties and thus have received considerable interest. We reviewed recent papers probing into the potential of nutrients with respect to lung function preservation and prevention of lung cancer risk, and suggest several hypothetical intervention patterns. Intakes of vitamins (i.e., A, C, D, E, B12), carotenoids, flavonoids, curcumins, resveratrol, magnesium, and omega-3 fatty acids all show protective effects against lung function loss, some mainly by improving average lung function and others through reducing decline rate. Dietary interventions early in life may help lung function reserve over the lifespan. Protective nutrient interventions among smokers are likely to mitigate the effects of cigarettes on lung health. We also discuss their underlying mechanisms and some possible causes for the inconsistent results in observational studies and supplementation trials. The role of the lung microbiome on lung health and its potential utility in identifying protective nutrients are discussed as well. More prospective cohorts and well-designed clinical trials are needed to promote the transition of individualized nutrient interventions into health policy.


A Unified Interactive Model Evaluation for Classification, Object Detection, and Instance Segmentation in Computer Vision.
Existing model evaluation tools mainly focus on evaluating classification models, leaving a gap in evaluating more complex models, such as object detection. In this paper, we develop an open-source visual analysis tool, Uni-Evaluator, to support a unified model evaluation for classification, object detection, and instance segmentation in computer vision. The key idea behind our method is to formulate both discrete and continuous predictions in different tasks as unified probability distributions. Based on these distributions, we develop 1) a matrix-based visualization to provide an overview of model performance; 2) a table visualization to identify the problematic data subsets where the model performs poorly; 3) a grid visualization to display the samples of interest. These visualizations work together to facilitate the model evaluation from a global overview to individual samples. Two case studies demonstrate the effectiveness of Uni-Evaluator in evaluating model performance and making informed improvements.


[Recurrent erysipelas].
Erysipelas is clinically defined as a febrile skin infection with a sudden onset of a red indurated expanding plaque with distinct border. A typical erysipelas presents various degrees of cutaneous erythema, oedema and is characterized by less well-defined margins. The responsible agent can be a beta-hemolytic streptococcus (group A), Staphylococcus aureus and other streptococci (group B, C and D). Common predisposing factors are venous insufficiency, lymphatic obstruction and underlying illnesses. Guidelines for the diagnosis and the management of erysipelas are proposed. The episodes of erysipelas ordinarily respond promptly to penicillin or a beta-lactamase resistant antibiotic. Prevention of recurrent erysipelas by means of an antibiotic prophylaxis is discussed.


Two-monoclonal-antibody sandwich-type assay for thyrotropin, with use of an avidin-biotin separation technique.
We have developed a sensitive, specific, noncompetitive, sandwich-type radioimmunoassay for human thyrotropin (hTSH), which can be performed in 30 min. The assay involves two monoclonal antibodies, selected for high affinity and specificity and also for reaction against antigenic sites on hTSH that are distal from each other. One of these antibodies is labeled with 125I; the other is conjugated covalently to biotin. Polystyrene beads were also conjugated covalently to biotin. After conjugation, the beads were incubated with avidin. These beads represent a rapid, simple method for separating hTSH-bound antibody from free antibody. The biotin-antibody-hTSH-125I-labeled antibody complexes bind to the beads and hTSH concentration is directly related to counts per minute. This assay can detect hTSH at a concentration of 0.06 milli-unit/L in serum.


A consultation-level intervention to improve care of frequently attending patients: a cluster randomised controlled feasibility trial.
Frequent attenders (FAs) to primary care receive considerable NHS resources without necessarily gaining benefit, and may even be harmed. To assess the feasibility of a consultation-level intervention to improve care and address service use of FAs. A cluster randomised controlled feasibility trial was undertaken. The study used a mixed-methods process evaluation and took place in six practices in England. All practices screened the top 3% of all attending patients over the previous 12 months for eligibility. Following randomisation, intervention patients were matched with named GPs, trained to use the Background, Affect, Trouble, Handling, Empathy (BATHE) technique during consultations. Telephone consultations were encouraged. Feasibility outcomes assessed were recruitment, retention, data collection and completeness, implementation fidelity, and acceptability. A total of 599/1328 (45.1%) FAs were eligible. Four practices were randomised to the intervention (n = 451) and two to usual care (n = 148). A total of 96 (23.7%) patients were recruited to complete questionnaires. Retention and completeness of data were good; for example, 76% of those agreeing to complete questionnaires did so at the 12-month assessment point. Thirty-four GPs were trained and delivered BATHE ≥1 times to 50.1% of patients (n = 577 consultations). There were minimal increases in continuity and telephone consultations. Patients were positive about the intervention, but noticed little change in their care. Despite valuing BATHE, low adherence to training was indicated and GPs used it less than anticipated. It was feasible to identify FAs and collect trial data. GPs were keen to engage and there was evidence that the BATHE technique was taken into practice. Optimising training is likely to improve fidelity. The intervention was low cost and low risk.


A dissipative particle dynamics simulation of controlled loading and responsive release of theranostic agents from reversible crosslinked triblock copolymer vesicles.
To control the transport stability and release efficiency of loaded theranostic drugs in triblock copolymer carriers, the reversible crosslinking ability is of great significance. A molecular level exploration of such a function is needed to extend existing stabilizing and responsive dissociation mechanisms of carriers. Here, dissipative particle dynamics simulations were used to first demonstrate the formation of triblock copolymer vesicular carriers. Chemical crosslinking was used to strengthen the structural stability of the vesicle shell to avoid drug leakage. Reversible decrosslinking along with dissociation of the vesicle and release of loaded drugs were then explored. The structural, energetic and dynamical properties of the system were discussed at the molecular level. The regulation mechanism of drug release patterns was revealed by systematically exploring the effect of intra and intermolecular repulsive interactions. The results indicate that the chemical crosslinking of copolymers enhanced the compactness of the vesicle shell with a strengthened microstructure, increased binding energy, and limited chain migration, thus achieving more stable delivery of drugs. In terms of drug release, we clarified how the pairwise interactions of beads in the solution system affect the responsive dissociation of the vesicle and associated release patterns (speed and amount) of drugs. More efficient delivery and smart release of theranostic drugs are achieved using such reversible crosslinked triblock copolymer vesicles.


Lipopeptisomes: Anticancer peptide-assembled particles for fusolytic oncotherapy.
Anticancer peptides (ACPs) are cationic amphiphiles that preferentially kill cancer cells through folding-dependent membrane disruption. Although ACPs represent attractive therapeutic candidates, particularly against drug-resistant cancers, their successful translation into clinical practice has gone unrealized due to their poor bioavailability, serum instability and, most importantly, severe hemolytic toxicity. Here, we exploit the membrane-specific interactions of ACPs to prepare a new class of peptide-lipid particle, we term a lipopeptisome (LP). This design sequesters loaded ACPs within a lipid lamellar corona to avoid contact with red blood cells and healthy tissues, while affording potent lytic destruction of cancer cells following LP-membrane fusion. Biophysical studies show ACPs rapidly fold at, and integrate into, liposomal membranes to form stable LPs with high loading efficiencies (>80%). Rational design of the particles to possess lipid combinations mimicking that of the aberrant cancer cell outer leaflet allows LPs to rapidly fuse with tumor cell membranes and afford localized assembly of loaded ACPs within the bilayer. This leads to preferential fusolytic killing of cancer cells with minimal collateral toxicity towards non-cancerous cells and erythrocytes, thereby imparting clinically relevant therapeutic indices to otherwise toxic ACPs. Thus, integration of ACPs into self-assembled LPs represents a new delivery strategy to improve the therapeutic utility of oncolytic agents, and suggests this technology may be added to targeted combinatorial approaches in precision medicine. STATEMENT OF SIGNIFICANCE: Despite their significant clinical potential, the therapeutic utility of many ACPs has been limited by their collateral hemolysis during administration. Leveraging the membrane-specific interactions of ACPs, here we prepare self-assembled peptide-lipid nanoparticles, or 'lipopeptisomes' (LPs), capable of preferentially fusing with and lysing cancer cell membranes. Key to this fusolytic action is the construction of LPs from lipids simulating the cancer cell outer leaflet. This design recruits the oncolytic peptide payload into the carrier lamella and allows for selective destruction of cancer cells without disrupting healthy cells. Consequently, LPs impart clinically relevant therapeutic indexes to previously toxic ACPs, and thus open new opportunities to improve the clinical translation of oncolytics challenged by narrow therapeutic windows.


Depletion of normal B cells with rituximab as an adjunct to IL-2 therapy for renal cell carcinoma and melanoma.
We postulated that in patients with metastatic renal cell carcinoma (RCC) or melanoma, depletion of normal B cells using the anti-CD20 mAb rituximab before treatment with low-dose interleukin (IL)-2 would improve clinical outcome. Rituximab (375 mg/m(2)) weekly for 4 weeks. IL-2 [11 (million units) daily] s.c., 4 days a week for weeks 5-8, followed by a 2-week rest (weeks 9 and 10). Patients without disease progression continued on IL-2. Disease re-evaluation was performed after rituximab and after every course of IL-2. Fifteen patients with RCC and six with melanoma were enrolled. One patient had a partial response and seven patients had stable disease. Toxicities were similar to those expected with IL-2 alone, and there were no grade 4 events. Circulating B cells were depleted in all patients. The subsequent low-dose IL-2 increased absolute numbers of natural killer cells, activated CD4(+) and activated CD8(+) T cells. Expanded T cells produced interferon-gamma, but not IL-4. Proliferation of peripheral blood lymphocytes to phytohemagglutinin was diminished following rituximab treatment, suggesting that B cells participate in this response in vitro. Our results suggest that depletion of circulating B cells with rituximab does not increase the response rate, alter the toxicity profile or change the biological activity in response to low-dose IL-2 in patients with RCC or melanoma.


Two non-parametric methods for derivation of constraints from radiotherapy dose-histogram data.
Dose constraints based on histograms provide a convenient and widely-used method for informing and guiding radiotherapy treatment planning. Methods of derivation of such constraints are often poorly described. Two non-parametric methods for derivation of constraints are described and investigated in the context of determination of dose-specific cut-points-values of the free parameter (e.g., percentage volume of the irradiated organ) which best reflect resulting changes in complication incidence. A method based on receiver operating characteristic (ROC) analysis and one based on a maximally-selected standardized rank sum are described and compared using rectal toxicity data from a prostate radiotherapy trial. Multiple test corrections are applied using a free step-down resampling algorithm, which accounts for the large number of tests undertaken to search for optimal cut-points and the inherent correlation between dose-histogram points. Both methods provide consistent significant cut-point values, with the rank sum method displaying some sensitivity to the underlying data. The ROC method is simple to implement and can utilize a complication atlas, though an advantage of the rank sum method is the ability to incorporate all complication grades without the need for grade dichotomization.


The expression of overexpressed PTEN enhanced IR-induced apoptosis of myocardial cells.
Myocardial cell apoptosis is an important pathologic basis of ischemia-reperfusion injury (I/R). PI3K/Akt signaling pathway involves in cell growth, survival, and apoptosis regulation, thus playing an important role in the protection of I/R injury. PTEN is a negative regulatory factor of PI3K/Akt signaling pathway. This study established rat I/R injury model after AMI and myocardial cell I/R injury model to explore the regulatory role of PTEN-PI3K/Akt signaling pathway in myocardial I/R injury in vivo and in vitro. Rat myocardial I/R injury model was established. PTEN and p-Akt expressions in myocardial tissue were compared. H9C2 cells were incubated in I/R condition for 12 h, followed by reoxygenation for 12 h. H9C2 cells were divided into three groups, including I/R+pSicoR-Blank, I/R+pSicoR-PTEN, and I/R+pSicoR-PTEN+VO-Ohpic. PTEN, p-Akt, Bcl-2, and Bax expressions were detected. Cell apoptosis was measured by flow cytometry. PTEN expression significantly increased, while p-Akt level markedly declined in myocardial tissue in I/R group compared with Sham group. Temporary PTEN downregulation and p-Akt elevation appeared at 2 h after I/R. I/R treatment markedly enhanced PTEN and Bax expressions, increased cell apoptosis, and reduced p-Akt and Bcl-2 levels. PTEN overexpression significantly enhanced Bax expression and cell apoptosis, while declined p-Akt and Bcl-2 in H9C2 after I/R. PTEN inhibited by VO-Ohpic markedly downregulated p-Akt and Bcl-2 expressions, whereas reduced Bax level and cell apoptosis. The overexpression of PTEN aggravated myocardial cell apoptosis after I/R. The blockage of PTEN enhanced PI3K/Akt signaling pathway and attenuated cell apoptosis induced by I/R.


BEX5/RabA1b regulates trans-Golgi network-to-plasma membrane protein trafficking in Arabidopsis.
Constitutive endocytic recycling is a crucial mechanism allowing regulation of the activity of proteins at the plasma membrane and for rapid changes in their localization, as demonstrated in plants for PIN-FORMED (PIN) proteins, the auxin transporters. To identify novel molecular components of endocytic recycling, mainly exocytosis, we designed a PIN1-green fluorescent protein fluorescence imaging-based forward genetic screen for Arabidopsis thaliana mutants that showed increased intracellular accumulation of cargos in response to the trafficking inhibitor brefeldin A (BFA). We identified bex5 (for BFA-visualized exocytic trafficking defective), a novel dominant mutant carrying a missense mutation that disrupts a conserved sequence motif of the small GTPase, RAS GENES FROM RAT BRAINA1b. bex5 displays defects such as enhanced protein accumulation in abnormal BFA compartments, aberrant endosomes, and defective exocytosis and transcytosis. BEX5/RabA1b localizes to trans-Golgi network/early endosomes (TGN/EE) and acts on distinct trafficking processes like those regulated by GTP exchange factors on ADP-ribosylation factors GNOM-LIKE1 and HOPM INTERACTOR7/BFA-VISUALIZED ENDOCYTIC TRAFFICKING DEFECTIVE1, which regulate trafficking at the Golgi apparatus and TGN/EE, respectively. All together, this study identifies Arabidopsis BEX5/RabA1b as a novel regulator of protein trafficking from a TGN/EE compartment to the plasma membrane.


Stiffness modification of two ankle-foot orthosis types to optimize gait in individuals with non-spastic calf muscle weakness - a proof-of-concept study.
To reduce gait problems in individuals with non-spastic calf muscle weakness, spring-like ankle-foot orthoses (AFOs) are often applied, but they are not individually optimized to treatment outcome. The aim of this proof-of-concept study was to evaluate the effects of modifying the stiffness for two spring-like AFO types with shoes-only as reference on gait outcomes in three individuals with calf muscle weakness due to polio. We assessed 3D gait biomechanics, walking speed and walking energy cost for shoes-only and five stiffness conditions of a dorsal-leaf-spring AFO and a spring-hinged AFO. Outcomes were compared between stiffness conditions in the two AFOs and three subjects. Maximum ankle dorsiflexion angle decreased with increasing stiffness in both AFOs (up to 6-8°) and all subjects. Maximum knee extension angle changed little between stiffness conditions, however different responses between the AFOs and subjects were observed compared to shoes-only. Walking speed remained unchanged across conditions. For walking energy cost, we found fairly large differences across stiffness conditions with both AFOs and between subjects (range 3-15%). Modifying AFO stiffness in individuals with non-spastic calf muscle weakness resulted in substantial differences in ankle biomechanics and walking energy cost with no effect on speed. Our results provide proof-of-concept that individually optimizing AFO stiffness can clinically beneficially improve gait performance.


Changes in fruit and vegetable consumption habits from pre-pregnancy to early pregnancy among Norwegian women.
A healthy diet is important for pregnancy outcome and the current and future health of woman and child. The aims of the study were to explore the changes from pre-pregnancy to early pregnancy in consumption of fruits and vegetables (FV), and to describe associations with maternal educational level, body mass index (BMI) and age. Healthy nulliparous women were included in the Norwegian Fit for Delivery (NFFD) trial from September 2009 to February 2013, recruited from eight antenatal clinics in southern Norway. At inclusion, in median gestational week 15 (range 9-20), 575 participants answered a food frequency questionnaire (FFQ) where they reported consumption of FV, both current intake and recollection of pre-pregnancy intake. Data were analysed using a linear mixed model. The percentage of women consuming FV daily or more frequently in the following categories increased from pre-pregnancy to early pregnancy: vegetables on sandwiches (13 vs. 17%, p <0.01), other vegetables (11 vs. 14%, p = 0.01), fruits (apples, pears, oranges or bananas) (24 vs. 41%, p < 0.01), other fruits and berries (8 vs. 15%, p < 0.01) and fruits and vegetables as snacks (14 vs. 28%, p < 0.01). The percentage of women who reported at least daily consumption of vegetables with dinner (22% at both time points) was stable. A higher proportion of older women increased their consumption of vegetables and fruits as snacks from pre-pregnancy to early pregnancy compared to younger women (p=0.04). We found an increase in the proportion of women consuming FV daily or more frequently from pre-pregnancy to early pregnancy. ClinicalTrials.gov database, NCT01001689 . https://clinicaltrials.gov/ct2/show/NCT01001689?term=NCT01001689&rank=1 .


Unlocking the catalytic potential of tris(3,4,5-trifluorophenyl)borane with microwave irradiation.
The catalytic activity of tris(3,4,5-trifluorophenyl)borane has been explored in the 1,2-hydroboration reactions of unsaturated substrates. Under conventional conditions, the borane was found to be active only in the hydroboration of aldehyde, ketone and imine substrates, with alkenes and alkynes not being reduced effectively. The use of microwave irradiation on the other hand has permitted alkenes and alkynes to be hydroborated in good yields.


Human erythrocyte band 3 is a host receptor for Plasmodium falciparum glutamic acid-rich protein.
Malaria remains a major global threat to human health and economic development. Microvascular lesions caused by Plasmodium falciparum-infected human erythrocytes/red blood cells are hallmarks of severe pathogenesis contributing to high mortality, particularly in children from sub-Saharan Africa. In this study, we used a phage display complementary DNA library screening strategy to identify P falciparum glutamic acid-rich protein (PfGARP) as a secreted ligand that recognizes an ectodomain of human erythrocyte anion-exchanger, band 3/AE1, as a host receptor. Domain mapping of PfGARP revealed distinct nonoverlapping repeats encoding the immune response epitopes and core erythrocyte-binding activity. Synthetic peptides derived from the erythrocyte-binding repeats of PfGARP induced erythrocyte aggregation reminiscent of the rosetting phenomenon. Using peptides derived from the immunogenic repeats, a quantitative immunoassay was developed to detect a selective immune response against PfGARP in human plasma samples obtained from patients in rural Mali, suggesting the feasibility of PfGARP as a potential biomarker of disease progression. Collectively, our results suggest that PfGARP may play a functional role in enhancing the adhesive properties of human erythrocytes by engaging band 3 as a host receptor. We propose that immunological and pharmacological inhibition of PfGARP may unveil new therapeutic options for mitigating lesions in cerebral and pregnancy-associated malaria.


Seeing the Hidden Lamina: Effects of Exsanguination on the Optic Nerve Head.
To introduce an experimental approach for direct comparison of the primate optic nerve head (ONH) before and after death by exsanguination. The ONHs of four eyes from three monkeys were imaged with spectral-domain optical coherence tomography (OCT) before and after exsanguination under controlled IOP. ONH structures, including the Bruch membrane (BM), BM opening, inner limiting membrane (ILM), and anterior lamina cribrosa (ALC) were delineated on 18 virtual radial sections per OCT scan. Thirteen parameters were analyzed: scleral canal at BM opening (area, planarity, and aspect ratio), ILM depth, BM depth; ALC (depth, shape index, and curvedness), and ALC visibility (globally, superior, inferior, nasal, and temporal quadrants). All four ALC quadrants had a statistically significant improvement in visibility after exsanguination (overall P < 0.001). ALC visibility increased by 35% globally and by 36%, 37%, 14%, and 4% in the superior, inferior, nasal, and temporal quadrants, respectively. ALC increased 4.1%, 1.9%, and 0.1% in curvedness, shape index, and depth, respectively. Scleral canals increased 7.2%, 25.2%, and 1.1% in area, planarity, and aspect ratio, respectively. ILM and BM depths averaged -7.5% and -55.2% decreases in depth, respectively. Most, but not all, changes were beyond the repeatability range. Exsanguination allows for improved lamina characterization, especially in regions typically blocked by shadowing in OCT. The results also demonstrate changes in ONH morphology due to the loss of blood pressure. Future research will be needed to determine whether there are differences in ONH biomechanics before and after exsanguination and what those differences would imply.


Changes Introduced in the Open Reading Frame of Bovine Viral Diarrhea Virus During Serial Infection of Pregnant Swine.
Bovine viral diarrhea virus (BVDV) is one of the most economically important viruses of cattle, but this pathogen is also able to infect pigs, camelids, and a wide range of domestic and wild ruminants. BVDV isolates circulating in animal populations are genetically and antigenically highly diverse. Acute BVDV infections in cattle cause the introduction of many substitutions in the viral genome. Serial infection of pregnant sheep with a BVDV-1b isolate of bovine origin was also associated with great numbers of substitutions. To our knowledge, genomic changes arising during BVDV infections in swine have not been investigated. The purpose of this study was to investigate the changes occurring in the open reading frame (ORF) of BVDV during serial infection of pregnant swine with a BVDV isolate of bovine origin. The BVDV-1b isolate AU526 was serially passaged in six pregnant gilts, two of which gave birth to live piglets congenitally infected with BVDV. The complete ORF sequences of 14 BVDV isolates obtained from pregnant gilts and their piglets were determined. Their analysis revealed that serial transmission of AU526 in pregnant swine resulted in many genomic changes. All isolates of porcine origin shared 32 nucleotide and 12 amino acid differences with the virus inoculum AU526. These changes were detected after a single passage in pregnant swine and were conserved during the subsequent five passages. Amino acid changes occurred primarily in genomic regions encoding the BVDV structural proteins E2 and E rns . These results suggest that BVDV infections in pregnant swine may contribute significantly to the genetic variability of BVDV and lead to the appearance of adaptive changes.


Evaluation of a modified sphincter pharyngoplasty in the treatment of speech problems due to palatal insufficiency.
Results are presented of the first 10 children treated with a sphincter-type pharyngoplasty for speech defects due to velopharyngeal insufficiency. Seven of these children had cleft palates, 4 of the submucous variety. The operation, modified from one devised by Orticochea, is described in detail and emphasizes the importance of sphincter action closure of the nasopharynx. We believe this closure must be at a very high level. Preoperative and postoperative assessments of the patients by 4 observers are detailed in depth. Eight of the 10 patients showed improvement and none were made worse. We believe this operation, which places a functioning sphincter near the optimum site, is of considerable value in the treatment of selected patients suffering speech defects resulting from velopharyngeal incompetence.


Surgery for chronic obstructive pulmonary disease: the place for lung volume reduction and transplantation.
Lung volume reduction surgery and lung transplantation have been shown to improve lung function, exercise capacity, and quality of life in patients with advanced emphysema. Because the indications for both surgical procedures overlap, lung volume reduction surgery may be used as an alternative treatment or as a "bridge" to lung transplantation. In this article, we discuss patient selection, clinical outcome parameters, and the morbidity and mortality associated with each surgical procedure. We focus on the different preoperative predictors of good and poor outcomes after lung volume reduction surgery, the role of pulmonary rehabilitation, and the preferred surgical techniques for lung volume reduction surgery. An overview of the postoperative care of emphysema patients who undergo single-lung transplantation is also discussed.


Molecular Mechanisms of Organic Anion Transporting Polypeptide-Mediated Organic Anion Clearance at the Blood-Cerebrospinal Fluid Barrier.
The blood-cerebrospinal fluid barrier (BCSFB), formed by the choroid plexus epithelial (CPE) cells, plays an active role in removing drugs and metabolic wastes from the brain. Recent functional studies in isolated mouse choroid plexus (CP) tissues suggested the presence of organic anion transporting polypeptides (OATPs, encoded by SLCOs) at the apical membrane of BCSFB, which may clear large organic anions from the cerebrospinal fluid (CSF). However, the specific OATP isoform involved is unclear. Using quantitative fluorescence imaging, we showed that the fluorescent anions sulforhodamine 101 (SR101), fluorescein methotrexate (FL-MTX), and 8-fluorescein-cAMP (fluo-cAMP) are actively transported from the CSF to the subepithelial space in CP tissues isolated from wild-type mice. In contrast, transepithelial transport of these compounds across the CPE cells was abolished in Oatp1a/1b-/- mice due to impaired apical uptake. Using transporter-expressing cell lines, SR101, FL-MTX, and fluo-cAMP were additionally shown to be transported by mouse OATP1A5 and its human counterpart OATP1A2. Kinetic analysis showed that estrone-3-sulfate and SR101 are transported by OATP1A2 and OATP1A5 with similar Michaelis-Menten constants (Km). Immunofluorescence staining further revealed the presence of OATP1A2 protein in human CP tissues. Together, our results suggest that large organic anions in the CSF are actively transported into CPE cells by apical OATP1A2 (OATP1A5 in mice), then subsequently effluxed into the blood by basolateral multidrug resistance-associated proteins (MRPs). As OATP1A2 transports a wide array of endogenous compounds and xenobiotics, the presence of this transporter at the BCSFB may imply a novel clearance route for drugs and neurohormones from the CSF. SIGNIFICANCE STATEMENT: Drug transporters at the blood-cerebrospinal fluid (CSF) barrier play an important but understudied role in brain drug disposition. This study revealed a functional contribution of rodent organic anion transporting polypeptide (OATP) 1A5 towards the CSF clearance of organic anions and suggested a similar role for OATP1A2 in humans. Delineating the molecular mechanisms governing CSF organic anion clearance may help to improve the prediction of central nervous system (CNS) pharmacokinetics and identify drug candidates with favorable CNS pharmacokinetic properties.


Studies of the regulatory effects of the sex hormones on antibody formation and stem cell differentiation.
The primary and secondary immune responses to thymus-dependent and -independent antigens were evaluated in normal male and female mice and in castrated male mice. Both IgM antibody production in the primary response and IgG antibody production in the secondary response were enhanced in females vs. males of equivalent age. Castration of the male converted this animal to a female in terms of responsiveness to the thymus-dependent group of antigens, while inducing equivalent or even greater enhanced responsiveness over the female to the thymus-independent antigen, polyvinylpyrrolidone. Further characteristics of the changes in lymphoid organs were determined in the castrated animal vs. normal males and females. It was shown that the spleen and thymus became markedly hyperplastic, the organ weights exceeding the female, which in turn were greater than in the male. The enhanced weight of the thymus was shown to be due to increased numbers of cortisone-sensitive cells, the absolute number of cortisone-resistant cells remaining equivalent to normal males and females. Thus, the increased thymic weight of the female also resided in the cortisone-sensitive population. Peripheral lymphocyte counts in castrated animals exceeded both normal males and females. Further experiments in gonadectomized males provided evidence that increased thymic cell activity per se played a role in enhanced response to thymus-dependent antigens, but that a thymic-derived hormone mediated the enhanced effect to the thymus-independent antigen in the castrated animal. The capacity for loss of androgenic hormone-producing tissue to generate enhanced differentiation of stem cells was denoted by experiments in which numbers of spleen colonies and uptake of (59)Fe, employed as an index of hematopoiesis 1 wk after reconstitution of lethally irradiated castrated and normal recipients, were enhanced in gonadectomized male animals. Thus, in summary, changes in sex hormone levels exerted a marked influence on immune responsiveness and stem cell differentiation, by increasing numbers of functioning cells, by promoting cellular differentiation, as well as by promoting cellular function via hormonal effects.


Enhancement of tolerance against flonicamid in Solenopsis invicta (Hymenoptera: Formicidae) through overexpression of CYP6A14.
Solenopsis invicta is a main issue in southern China and is causing significant damage to the local ecological environment. The extensive use of insecticides has resulted in the development of tolerance in S. invicta. In our study, ten S. invicta colonies from Sichuan Province exhibited varying degrees of tolerance against flonicamid, with LC50 values from 0.49 mg/L to 8.54 mg/L. The sensitivity of S. invicta to flonicamid significantly increased after treatment with the P450 enzyme inhibitor piperonyl butoxide (PBO). Additionally, the activity of P450 in S. invicta was significantly enhanced after being treated with flonicamid. Flonicamid induced the expression levels of CYP4aa1, CYP9e2, CYP4C1, and CYP6A14. The expression levels of these P450 genes were significantly higher in the tolerant colonies compared to the sensitive colonies, and the relative copy numbers of CYP6A14 in the tolerant colonies were 2.01-2.15 fold. RNAi feeding treatment effectively inhibited the expression of P450 genes, thereby reducing the tolerance of S. invicta against flonicamid. In addition, the overexpression of CYP6A14 in D. melanogaster resulted in reduced sensitivity to flonicamid. Our investigations revealed hydrophobic interactions between flonicamid and seven amino acid residues of CYP6A14, along with the formation of a hydrogen bond between Glu306 and flonicamid. Our findings suggest that flonicamid can effectively control S. invicta and P450 plays a pivotal role in the tolerance of S. invicta against flonicamid. The overexpression of CYP6A14 also increased tolerance to flonicamid.


The age of anxiety: role of animal models of anxiolytic action in drug discovery.
Anxiety disorders are common, serious and a growing health problem worldwide. However, the causative factors, aetiology and underlying mechanisms of anxiety disorders, as for most psychiatric disorders, remain relatively poorly understood. Animal models are an important aid in giving insight into the aetiology, neurobiology and, ultimately, the therapy of human anxiety disorders. The approach, however, is challenged with a number of complexities. In particular, the heterogeneous nature of anxiety disorders in humans coupled with the associated multifaceted and descriptive diagnostic criteria, creates challenges in both animal modelling and in clinical research. In this paper, we describe some of the more widely used approaches for assessing the anxiolytic activity of known and potential therapeutic agents. These include ethological, conflict-based, hyponeophagia, vocalization-based, physiological and cognitive-based paradigms. Developments in the characterization of translational models are also summarized, as are the challenges facing researchers in their drug discovery efforts in developing new anxiolytic drugs, not least the ever-shifting clinical conceptualization of anxiety disorders. In conclusion, to date, although animal models of anxiety have relatively good validity, anxiolytic drugs with novel mechanisms have been slow to emerge. It is clear that a better alignment of the interactions between basic and clinical scientists is needed if this is to change.