Autofluorescence properties of rat liver under hypermetabolic conditions.
Autofluorescence response to oxygen supply modulation has been investigated in livers of rats under the hypermetabolic state associated to a pathological condition-hyperthyroidism-that is known to enhance hepatocyte metabolic activities involving both NAD, i.e. oxidative pathways engaged in ATP synthesis, and NADP, i.e. reductive bio-synthesis and antioxidant functions. Experiments have been performed on rats in normal condition or submitted to long-term thyroxine (T(4)) administration. Histological inspection did not show any appreciable morphological alteration in liver parenchyma; biochemical analysis indicated an increase in both NADP(+) and NADPH contents. Autofluorescence properties have been monitored in vivo, via a fiber optic probe, on exposed livers both during induction of global ischemia and after restoration of blood circulation. Alteration of oxygen supply modulated liver autofluorescence properties, mainly as to NAD(P)H contribution, in dependence of changes in pyridine coenzymes redox state. With respect to euthyroid, hyperthyroid rat livers exhibited higher autofluorescence signals in all phases of the experiment, and a faster signal decay time upon reoxygenation. The results have been interpreted on the basis of a larger content of NADPH-the coenzyme not directly oxidized in respiratory processes and likely providing an almost constant autofluorescence background contribution-and of uncoupling effects facilitating the respiratory NADH oxidation, associated with the hyperthyroid condition. The results obtained in the liver hypermetabolic model provide interesting perspectives for a further improvement of the diagnostic implications of autofluorescence.


Endolysosomal Deficits Augment Mitochondria Pathology in Spinal Motor Neurons of Asymptomatic fALS Mice.
One pathological hallmark in ALS motor neurons (MNs) is axonal accumulation of damaged mitochondria. A fundamental question remains: does reduced degradation of those mitochondria by an impaired autophagy-lysosomal system contribute to mitochondrial pathology? We reveal MN-targeted progressive lysosomal deficits accompanied by impaired autophagic degradation beginning at asymptomatic stages in fALS-linked hSOD1(G93A) mice. Lysosomal deficits result in accumulation of autophagic vacuoles engulfing damaged mitochondria along MN axons. Live imaging of spinal MNs from the adult disease mice demonstrates impaired dynein-driven retrograde transport of late endosomes (LEs). Expressing dynein-adaptor snapin reverses transport defects by competing with hSOD1(G93A) for binding dynein, thus rescuing autophagy-lysosomal deficits, enhancing mitochondrial turnover, improving MN survival, and ameliorating the disease phenotype in hSOD1(G93A) mice. Our study provides a new mechanistic link for hSOD1(G93A)-mediated impairment of LE transport to autophagy-lysosomal deficits and mitochondrial pathology. Understanding these early pathological events benefits development of new therapeutic interventions for fALS-linked MN degeneration.


Determination of antiepileptic drugs.
The present paper reviews gas and liquid chromatographic methods for the determination of the most commonly monitored antiepileptic drugs: phenobarbital, phenytoin, carbamazepine, primidone, ethosuximide, valproic acid and clonazepam along with a new compound, progabide. The individual classes of drugs are first treated separately to highlight specific aspects of their quantification, and this is followed by an overview of those methods permitting the concomitant analysis of two or more antiepileptic compounds. Sample preparation techniques as well as comparisons between chromatographic and other techniques are treated more fully in separate sections.


Targeting vascular inflammation in ischemic stroke: Recent developments on novel immunomodulatory approaches.
Ischemic stroke is a devastating and debilitating medical condition with limited therapeutic options. However, accumulating evidence indicates a central role of inflammation in all aspects of stroke including its initiation, the progression of injury, and recovery or wound healing. A central target of inflammation is disruption of the blood brain barrier or neurovascular unit. Here we discuss recent developments in identifying potential molecular targets and immunomodulatory approaches to preserve or protect barrier function and limit infarct damage and functional impairment. These include blocking harmful inflammatory signaling in endothelial cells, microglia/macrophages, or Th17/γδ T cells with biologics, third generation epoxyeicosatrienoic acid (EET) analogs with extended half-life, and miRNA antagomirs. Complementary beneficial pathways may be enhanced by miRNA mimetics or hyperbaric oxygenation. These immunomodulatory approaches could be used to greatly expand the therapeutic window for thrombolytic treatment with tissue plasminogen activator (t-PA). Moreover, nanoparticle technology allows for the selective targeting of endothelial cells for delivery of DNA/RNA oligonucleotides and neuroprotective drugs. In addition, although likely detrimental to the progression of ischemic stroke by inducing inflammation, oxidative stress, and neuronal cell death, 20-HETE may also reduce susceptibility of onset of ischemic stroke by maintaining autoregulation of cerebral blood flow. Although the interaction between inflammation and stroke is multifaceted, a better understanding of the mechanisms behind the pro-inflammatory state at all stages will hopefully help in developing novel immunomodulatory approaches to improve mortality and functional outcome of those inflicted with ischemic stroke.


Human tumor xenografts treated with recombinant human tumor necrosis factor alone or in combination with interferons.
We have studied the activity of recombinant human tumor necrosis factor (rHuTNF) on six different human tumor xenografts derived from primary breast and bowel tumors and maintained by passage in nude mice. When 5 micrograms rHuTNF was given daily intratumorally to mice with established (approximately, 0.5 cm) tumors, total tumor regression was observed by 3-4 weeks in three of six xenograft lines. In a further two lines tumor stasis or significant slowing of growth was seen. This antitumor action was not accompanied by any consistent macroscopic change in the tumor such as necrosis, but histological examination revealed tumor cell degeneration and a large peritumoral infiltration of host inflammatory cells after 4-7 days therapy. In contrast to these data, little effect was seen when the same dose of rHuTNF was administered i.p. to nude mice bearing these tumors. In only two of six lines was any significant slowing of tumor growth seen. A 5-fold increase in the i.p. dose resulted in improved activity on only one of two xenograft lines tested. Efficacy of the i.p. rHuTNF dose could, however, be enhanced by simultaneous administration of human interferon, alpha or gamma. No obvious signs of toxicity were observed at all rHuTNF doses administered and weights of control and treated mice at the end of the experiments were comparable.


Patient-provider interaction, patient satisfaction, and health outcomes: testing explanatory models for people living with HIV/AIDS.
In the era of highly active antiretroviral therapy era, medication adherence and health-related quality of life (HRQOL) have become critical issues for people living with HIV/AIDS (PLWH). The purpose of this study was to test explanatory models of how patient-provider interaction and patient satisfaction are related to medication adherence and HRQOL for PLWH. A total of 344 PLWH receiving health-care services from a federally funded clinic in the southwest USA completed a survey questionnaire about their perception of interactions with providers, their satisfaction with services, their medication adherence, and their HRQOL. Comparing four latent variable structural equation models of direct and mediated effects of patient-provider interaction and patient satisfaction, the findings illustrate that the best model is one in which patient-provider interaction has a direct and positive effect on patient satisfaction, medication adherence, and HRQOL. These findings suggest that quality patient-provider interaction is a critical element of health-care services for PLWH, while patient satisfaction is an outcome measure and not a mediating factor for medication adherence and HRQOL.


Diffuse large B-cell non-Hodgkin lymphoma of the prostate presenting with urinary outlet obstruction: A case report.
Large B-cell non-Hodgkin lymphoma involving the prostate accounts for 0.09% and 0.1% of non-Hodgkin lymphoma. We report a case of prostatic large B-cell non-Hodgkin lymphoma in a 77-year-old male with symptoms of urinary retention most probably due to benign prostate hyperplasia. He underwent multiple needle core biopsies through transrectal ultrasound of the prostate. Histopathological examination of the core biopsies revealed diffuse infiltration by atypical lymphoid cells in the prostatic stroma, which was strongly positive for leukocyte common antigen and CD20. CD3, CK, PSA, BCL2, k-light chain, Cyclin D1 and synaptophysin were negative. Histopathology and immunohistochemical profile in the case was consistent with the diagnosis of diffuse large B-cell non-Hodgkin lymphoma of the prostate.


[Congenital vascular malformations in childhood].
The intracranial congenital vascular pathology, arterial, arteriovenous and venous, is reviewed, including the cavernomas as well. It is commented the possible origin, the clinical, histological and neuroradiological alterations as well as the possible treatment, when necessary, of every entity. The contribution of the magnetic resonance (MR), especially the angiographic MR (AMR) has been conclusive to the diagnosis. There have been considered six main groups which include the main entities: 1. Arterial anomalies; 2. Arteriovenous malformations; 3. Malformations of the Galen vein; 4. Venous malformations, and 5. Intracranial cavernomas.


Relationship of depressive symptoms with hospitalization and death in Japanese patients with heart failure.
Depressive symptoms are risk factors for poor outcomes and are positively associated with disease severity in patients with heart failure (HF). However, little is known about this association in the Japanese population. Therefore, we evaluated the prevalence of depressive symptoms and whether depressive symptoms predicted hospitalization for HF and death independent of disease severity and other factors in HF patients. A 2-year prospective cohort study was conducted on 115 outpatients with HF (73.9% males; mean age 64.7 years) in Tokyo. Of these, 27 patients (23.5%) were classified as having depressive symptoms (Center for Epidemiologic Studies Depression Scale score >or=16). Patients with depressive symptoms had higher rates of 2-year cardiac death or HF hospitalization (34.0% vs. 10.3%; P < .01), HF hospitalization (27.4% vs. 9.2%; P = .01), and all-cause death (27.4% vs. 7.2%; P < .01). Multivariate Cox regression analyses indicated that depressive symptoms were predictors of cardiac death or HF hospitalization (hazard ratio [HR], 3.29; P = .02), HF hospitalization (HR, 3.36; P = .04), and all-cause death (HR, 5.52; P = .01), independent of age and brain natriuretic peptide. Depressive symptoms were common and independent predictors of poor outcomes in Japanese patients with HF.


Chromatin immunoprecipitation and high-throughput sequencing from paraffin-embedded pathology tissue.
Formalin-fixed, paraffin-embedded (FFPE) samples represent the gold standard for storage of pathology samples. Here we describe pathology tissue chromatin immunoprecipitation (PAT-ChIP), a technique for extraction and high-throughput analysis, by techniques such as ChIP-seq, of chromatin derived from FFPE samples. Technically, the main challenge of PAT-ChIP is the preparation of good-quality chromatin from FFPE samples. Here we provide a detailed explanation of the methodology used, the choice of reagents and the troubleshooting steps required to establish a robust chromatin preparation procedure. Other steps have also been adapted from existing techniques to optimize their use for PAT-ChIP-seq. The protocol requires 4 d from the start to the end of the PAT-ChIP procedure. PAT-ChIP provides, for the first time, the chance to perform analyses of histone modifications and transcription factor binding on a genome-wide scale using patient-derived FFPE samples. This technique therefore allows the immediate use of pathology archives (even those that are several years old) for epigenetic analyses and the identification of candidate epigenetic biomarkers or targets.


Leishmanicidal Activity of Isoselenocyanate Derivatives.
Conventional chemotherapy against leishmaniasis includes agents exhibiting considerable toxicity. In addition, reports of drug resistance are not uncommon. Thus, safe and effective therapies are urgently needed. Isoselenocyanate compounds have recently been identified with potential antitumor activity. It is well known that some antitumor agents demonstrate effects against Leishmania In this study, the in vitro leishmanicidal activities of several organo-selenium and organo-sulfur compounds were tested against Leishmania major and Leishmania amazonensis parasites, using promastigotes and intracellular amastigote forms. The cytotoxicity of these agents was measured in murine peritoneal macrophages and their selectivity indexes were calculated. One of the tested compounds, the isoselenocyanate derivative NISC-6, showed selectivity indexes 2- and 10-fold higher than those of the reference drug amphotericin B when evaluated in L. amazonensis and L. major, respectively. The American strain (L. amazonensis) was less sensitive to NISC-6 than L. major, showing a trend similar to that observed previously for amphotericin B. In addition, we also observed that NISC-6 significantly reduced the number of amastigotes per infected macrophage. On the other hand, we showed that NISC-6 decreases expression levels of Leishmania genes involved in the cell cycle, such as topoisomerase-2 (TOP-2), PCNA, and MCM4, therefore contributing to its leishmanicidal activity. The effect of this compound on cell cycle progression was confirmed by flow cytometry. We observed a significant increase of cells in the G1 phase and a dramatic reduction of cells in the S phase compared to untreated cells. Altogether, our data suggest that the isoselenocyanate NISC-6 may be a promising candidate for new drug development against leishmaniasis.


h5-HT(1B) receptor-mediated constitutive Galphai3-protein activation in stably transfected Chinese hamster ovary cells: an antibody capture assay reveals protean efficacy of 5-HT.
1. Serotonin 5-HT(1B) receptors couple to G-proteins of the Gi/o family. However, their activation of specific G-protein subtypes is poorly characterised. Using an innovative antibody capture/guanosine-5'-0-(3-[(35)S]thio)-triphosphate ([(35)S]GTPgammaS) binding strategy, we characterised Galpha(i3) subunit activation by h5-HT(1B) receptors stably expressed in Chinese hamster ovary (CHO) cells. 2. The agonists, 5-HT, alniditan and BMS181,101, stimulated Galpha(i3), whereas methiothepin and SB224,289 behaved as inverse agonists. The selective 5-HT(1B) receptor ligand, S18127, modestly stimulated Galpha(i3) and reversed the actions of both 5-HT and methiothepin. S18127 (1 micro M) also produced parallel, dextral shifts of the 5-HT and methiothepin isotherms. 3. Isotopic dilution experiments ([(35)S]GTPgammaS versus GTPgammaS) revealed high-affinity [(35)S]GTPgammaS binding to Galpha(i3) subunits in the absence of receptor ligands indicating constitutive activity. High-affinity [(35)S]GTPgammaS binding was increased 2.8-fold by 5-HT with an increase in the affinity of GTPgammaS for Galpha(i3) subunits. In contrast, methiothepin halved the number of high-affinity binding sites and decreased their affinity. 4. h5-HT(1B) receptor-mediated Galpha(i3) subunit activation was dependent on the concentration of NaCl. At 300 mM, 5-HT stimulated [(35)S]GTPgammaS binding, basal Galpha(i3) activation was low and methiothepin was inactive. In contrast, at 10 mM NaCl, basal activity was enhanced and the inverse agonist activity of methiothepin was accentuated. Under these conditions, 5-HT decreased Galpha(i3) activation. 5. In conclusion, at h5-HT(1B) receptors expressed in CHO cells: (i) inverse agonist induced inhibition of Galpha(i3), and its reversal by S18127, reveals constitutive activation of this Galpha subunit; (ii) constitutive Galpha(i3) activation can be quantified by isotopic dilution [(35)S]GTPgammaS binding and (iii) decreasing NaCl concentrations enhances Galpha(i3) activation and leads to protean agonist properties of 5-HT: that is a switch to inhibition of Galpha(i3).


The changing relationship of obesity and disability, 1988-2004.
Recent studies suggest that the obese population may have been growing healthier since the 1960s, as indicated by a decrease in mortality and cardiovascular risk factors. However, whether these improvements have conferred decreased risk for disability is unknown. The obese population may be living longer with better-controlled risk factors but paradoxically experiencing more disability. To determine whether the association between obesity and disability has changed over time. Adults aged 60 years and older (N = 9928) with measured body mass index from 2 waves of the nationally representative National Health and Nutrition Examination Surveys (NHANES III [1988-1994] and NHANES 1999-2004). Reports of much difficulty or inability to perform tasks in 2 disability domains: functional limitations (walking one-fourth mile, walking up 10 steps, stooping, lifting 10 lb, walking between rooms, and standing from an armless chair) and activities of daily living (ADL) limitations (transferring, eating, and dressing). Among obese individuals, the prevalence of functional impairment increased 5.4% (from 36.8%-42.2%; P = .03) between the 2 surveys, and ADL impairment did not change. At time 1 (1988-1994), the odds of functional impairment for obese individuals were 1.78 times greater than for normal-weight individuals (95% confidence interval [CI], 1.47-2.16). At time 2 (1999-2004), this odds ratio increased to 2.75 (95% CI, 2.39-3.17), because the odds of functional impairment increased by 43% (OR 1.43; 95% CI, 1.18-1.75) among obese individuals during this period, but did not change among nonobese individuals. With respect to ADL impairment, odds for obese individuals were not significantly greater than for individuals with normal weight (OR, 1.31; 95% CI, 0.92-1.88) at time 1, but increased to 2.05 (95% CI, 1.45-2.88) at time 2. This was because the odds of ADL impairment did not change for obese individuals but decreased by 34% among nonobese individuals (OR, 0.66; 95% CI, 0.50-0.88). Recent cardiovascular improvements have not been accompanied by reduced disability within the obese older population. Rather, obese participants surveyed during 1999-2004 were more likely to report functional impairments than obese participants surveyed during 1988-1994, and reductions in ADL impairment observed for nonobese older individuals did not occur in those who were obese. Over time, declines in obesity-related mortality, along with a younger age at onset of obesity, could lead to an increased burden of disability within the obese older population.


Exploring symptom-level associations between anxiety and depression across developmental stages of adolescence: a network analysis approach.
Anxiety and depression often co-occur during adolescence, but the associations between symptoms of these two disorders in this developmental period are not yet fully understood. Network analysis provides a valuable approach to uncover meaningful associations among symptoms and offers insights for prevention and intervention strategies. This study aimed to investigate symptom-level associations between anxiety and depression using network analysis and to identify core symptoms, bridge symptoms, and differences in network structure across different stages of adolescence. The cross-sectional study was conducted in March 2022 in Shenzhen, China. Participants completed the Generalized Anxiety Disorder Scale-7 and Patient Health Questionnaire Depression Scale, along with demographic questionnaires assessing age and gender. Chinese adolescents aged 10 to 17 who were in Grades 5 or 6 of elementary school, Grades 1 or 2 of middle school, or Grades 1 or 2 of high school, and who could comprehensively understand and read Chinese were recruited as participants. Students in Grade 3 of middle and high schools were excluded due to their upcoming high school or college entrance examinations. Based on age, participants were categorized into early, middle, and late developmental stages of adolescence. "Loss of control" was among the most central symptoms in the comorbidity network throughout all three developmental stages; "excessive worry" and "anhedonia" emerged as the core symptoms in early adolescence, and "restlessness" as the core symptom in late adolescence. "Anhedonia," "sad mood," and "fatigue" were identified as bridge symptoms between anxiety and depression across all three developmental stages of adolescence. The global strength of the network in middle adolescence was significantly higher compared to the other two stages. These findings highlight the core and bridge symptoms that require special attention and intervention at each stage of adolescence. Moreover, significantly higher network connectivity in middle adolescence suggests this is a critical period for intervention to prevent the development of comorbid mental disorders.


Nebulized dexmedetomidine for attenuating hemodynamic response to laryngoscopy and endotracheal intubation in adult patients undergoing surgeries under general anaesthesia: a systematic review and meta-analysis of randomized controlled trials.
Sympathetic stimulation associated with laryngoscopy and endotracheal intubation (ETI) may lead to adverse cardio-/cerebro-vascular events in susceptible patients. Nebulization is a novel route for dexmedetomidine administration providing a large surface area for absorption while avoiding bradycardia and hypotension associated with intravenous route. We aimed to evaluate the efficacy and safety of dexmedetomidine nebulization for attenuating hemodynamic response to ETI in adult patients undergoing surgery under general anaesthesia. This systematic review was registered prospectively in the International Prospective Register of Systematic Reviews (CRD42023403624). PubMed, Embase (OvidSP), Cochrane library, Scopus (Elsevier), Web of Science (Clarivate) and Google Scholar were systematically searched from database inception until March 31, 2023. Two reviewers independently screened titles, abstracts and then full text against pre-specified eligibility criteria. Randomized controlled trials (RCTs) assessing effect of dexmedetomidine nebulization on hemodynamic response to ETI in adult patients undergoing surgeries under general anaesthesia were included. All studies reporting heart rate and systolic blood pressure at baseline and various time points after ETI were included. A pre-piloted data extraction form, Cochrane revised risk-of-bias tool (ROB 2) tool, GRADE approach and RevMan 5.4.1 (Cochrane Collaboration, Copenhagen, Denmark) were used for data extraction, risk of bias assessment, rating certainty of evidence and data synthesis respectively. Mean difference and relative risk with 95% Confidence Interval (CI) were used for continuous and dichotomous variables respectively. Six RCTs randomized 480 patients with ASA I/II patients aged < 60 years of age and undergoing elective surgeries to receive either dexmedetomidine (n = 240) or saline nebulization (n = 240). Except for one RCT which used 2 μg/kg, all other RCTs used dexmedetomidine dose of 1 μg /kg. Heart rate, systolic, diastolic and mean blood pressure were significantly lower in the dexmedetomidine group at all the measured time points after laryngoscopy and ETI with the only exception being systolic blood pressure at 3 min [mean difference -13.86 (95% CI -30.01 to 2.99), p = 0.09]. Bradycardia and hypotension as adverse effects were absent across the included studies. However, only one-third of the included studies had a low risk of bias and strength of evidence was very low according to the GRADE assessment. Compared to placebo, premedication with dexmedetomidine nebulization was associated with lower HR and BP following ETI without any risk of bradycardia and hypotension. However, the strength of evidence was very poor and came from just one country. Future well designed and conducted studies in different populations are warranted. PROSPERO Registration number: CRD42023403624.


Mouse genetic models for temporomandibular joint development and disorders.
The temporomandibular joint (TMJ) is a synovial joint essential for hinge and sliding movements of the mammalian jaw. Temporomandibular joint disorders (TMD) are dysregulations of the muscles or the TMJ in structure, function, and physiology, and result in pain, limited mandibular mobility, and TMJ noise and clicking. Although approximately 40-70% adults in the USA have at least one sign of TMD, the etiology of TMD remains largely unknown. Here, we highlight recent advances in our understanding of TMD in mouse models.


Progress in the development of enzyme-based nerve agent bioscavengers.
Acetylcholinesterase is the physiological target for acute toxicity of nerve agents. Attempts to protect acetylcholinesterase from phosphylation by nerve agents, is currently achieved by reversible inhibitors that transiently mask the enzyme active site. This approach either protects only peripheral acetylcholinesterase or may cause side effects. Thus, an alternative strategy consists in scavenging nerve agents in the bloodstream before they can reach acetylcholinesterase. Pre- or post-exposure administration of bioscavengers, enzymes that neutralize and detoxify organophosphorus molecules, is one of the major developments of new medical counter-measures. These enzymes act either as stoichiometric or catalytic bioscavengers. Human butyrylcholinesterase is the leading stoichiometric bioscavenger. Current efforts are devoted to its mass production with care to pharmacokinetic properties of the final product for extended lifetime. Development of specific reactivators of phosphylated butyrylcholinesterase, or variants with spontaneous reactivation activity is also envisioned for rapid in situ regeneration of the scavenger. Human paraoxonase 1 is the leading catalytic bioscavenger under development. Research efforts focus on improving its catalytic efficiency toward the most toxic isomers of nerve agents, by means of directed evolution-based strategies. Human prolidase appears to be another promising human enzyme. Other non-human efficient enzymes like bacterial phosphotriesterases or squid diisopropylfluorophosphatase are also considered though their intrinsic immunogenic properties remain challenging for use in humans. Encapsulation, PEGylation and other modifications are possible solutions to address this problem as well as that of their limited lifetime. Finally, gene therapy for in situ generation and delivery of bioscavengers is for the far future, but its proof of concept has been established.


Molecular dynamics simulations of EPON-862/DETDA epoxy networks: structure, topology, elastic constants, and local dynamics.
Structural, topological, mechanical and dynamical properties of EPON-862/DETDA epoxy networks are investigated with Molecular Dynamics (MD) simulations. The epoxy networks are composed of the resin Diglycidyl Ether Bisphenol F (DGEBF), also known as EPON-862, and the hardener Diethyl Toluene Diamine (DETDA). Systems with four different crosslinking degrees are examined; the effect of the degree of crosslinking on studied properties is thus determined. The computed quantities are retrieved by employing several simulation strategies and numerical methods of statistical mechanics in order to gain a rigorous and solid understanding of the aforementioned properties as well as to assess the accuracy and applicability of the methods employed. We quantify and analyze the local structure of the EPON-862/DETDA epoxy networks through the partial pair distribution functions, the Faber-Ziman partial structure factors and through simulated X-ray diffraction patterns, demonstrating good agreement with an experimental spectrum from a similar epoxy resin. The topology of the networks is examined with the aim of assessing percolation of connectivity, the properties of network fragments (subnetworks), and the distribution of functionalities of the crosslinks. The elastic constants of the systems are retrieved by employing two equilibrium (analysis of volume fluctuations, Parrinello-Rahman strain fluctuation relation) and one nonequilibrium (uniaxial tension/compression deformations at prescribed rate) method. Finally, the glass temperatures of the systems are estimated by calculating the density as a function of temperature and by analyzing the reorientational dynamics of bond vectors which describe relaxation processes at the segment level.


The purification and some properties of H-lysin from Aeromonas salmonicida.
H-lysin from Aeromonas salmonicida has been purified 1770-fold by freeze fractionation, ammonium sulphate precipitation, ion exchange chromatography and gel filtration chromatography. The purified material was predominantly H-lysin, devoid of detectable T-lysin, caseinase or gelatinase activity, although glycerophospholipid: cholesterol acyltransferase (GCAT) activity was present. The results suggested that H-lysin and GCAT activities were due to different extracellular products. Studies of the kinetics of haemolysis indicated that the H-lysin had an enzymic mode of action, and that initial erythrocyte damage appeared to precede lysis of the cell. The H-lysin was lethal to cultured rainbow trout gonad cells and leucocytes, but when it was injected intravenously in rainbow trout no pathological effects were observed.


The deformability of BHK cells and polyoma virus-transformed BHK cells in relation to locomotory behaviour.
Several aspects of the behaviour of polyoma virus-transformed BHK cells in culture have suggested that they are more deformable than BHK cells. This possibility was tested by applying negative pressure at the cell surface by means of a micropipette. It was found that PyBHK cells in early mitosis are twice as deformable as BHK cells in the same stage. In addition, the taut, non-ruffling margins of both cell types when fully spread are much less deformable than the extending, ruffling leading lamella. The degree of deformability of these cells is correlated with the distribution and organization of microfilaments and consistent with this, deformability increases greatly in the presence of cytochalasin B. The significance of deformability studies such as these is discussed.


Total hip replacement for the treatment of atraumatic slipped femoral capital epiphysis in dogs.
To evaluate dogs diagnosed with slipped femoral capital epiphysis which were treated by total hip replacement. Clinical and radiographic records of consecutive cases of slipped femoral capital epiphysis treated by total hip replacement were reviewed. Longer-term follow-up was by owner questionnaire. Fifteen cases were identified; 14 were affected unilaterally, 1 bilaterally. Median age at presentation was 13 months (range 6 to 30); median weight was 35 kg (range 10 to 66). Radiographically, proximal femoral sclerosis was evident in 13 of 16 hips. Thirteen hybrid and three cemented total hip replacements were performed. One dog had a stem complication five months postoperatively necessitating explantation. All other total hip replacements remained functional. At four weeks postoperatively lameness and pain scores were improved in all cases. Fourteen total hip replacements were examined after three months. Lameness was mild in 2 limbs and absent in 12; pain on manipulation was not apparent in any case. Significant radiographic complications were not encountered. Telephone questionnaires were performed for 14 cases (15 total hip replacements) at a median of 22 months postoperatively (range 8 to 45). Lameness was reportedly absent for 12 limbs and intermittent in 3. All owners felt that their dogs' quality of life was good. Total hip replacement can be a successful technique to treat slipped femoral capital epiphysis.


Disposition of 5-methyltetrahydrohomofolate and methotrexate in rats.
The pharmacologic disposition of [methyl-14C]5-methyltetrahydrohomofolate (MTHHF) and [3H]methotrexate (MTX) has been studied in rats after iv doses of 25 mg/kg. Thin-layer chromatography and high-pressure liquid chromatography were used to separate the parent compounds from possible products. In rat serum, levels of MTHHF decreased with half-lives of 7 and 55 mins; for MTX, the corresponding values of half-lives were 14 and 54 mins. No metabolites of MTHHF or MTX were found in the samples assayed. At 3, 6, and 24 hrs after administration of MTHHF or MTX, liver, kidney, and small intestine contained concentrations of the drugs greater than those in serum. In rats with cannulated bile ducts, 16% of 60% of the doses of MTHHF and MTX, respectively, were excreted in the bile within 4 hrs. In 24 hrs, 51% of the dose of MTHHF and 41% of the dose of MTX were excreted in the urine of rats without bile cannulas. During the same time, fecal excretion accounted for 21% of the dose of MTHHF and 17% of the dose of MTX. The data indicate that enterohepatic circulation is probably extensive for MTX but less so for MTHHF.


Developmental Changes of Muscle Synergies in an Infant's Walking.
Primitive stepping is one of the primitive reflexes in newborns in response to external stimuli. It is known that primitive stepping disappears about two months after birth, but its role and the relationship with voluntary gait acquired later are still unknown. In this study, we extracted muscle synergies, spatiotemporal coordination patterns of muscle activities, from EMG measured in one infant during growth (4-18 weeks of age) using non-negative matrix factorization (NMF). We found that a synergy changed before and after the disappearance of the primitive stepping, and others maintained the recruited muscles but changed the onset timing of the activations.


Modelling the fate of nitrite in an urbanized river using experimentally obtained nitrifier growth parameters.
Maintaining low nitrite concentrations in aquatic systems is a major issue for stakeholders due to nitrite's high toxicity for living species. This study reports on a cost-effective and realistic approach to study nitrite dynamics and improve its modelling in human-impacted river systems. The implementation of different nitrifying biomasses to model riverine communities and waste water treatment plant (WWTP)-related communities enabled us to assess the impact of a major WWTP effluent on in-river nitrification dynamics. The optimal kinetic parameters and biomasses of the different nitrifying communities were determined and validated by coupling laboratory experiments and modelling. This approach was carried out in the Seine River, as an example of a large human-impacted river with high nitrite concentrations. The simulation of nitrite fate was performed at a high spatial and temporal resolution (Δt = 10 min, dx¯ = 500 m) including water and sediment layers along a 220 km stretch of the Seine River for a 6-year period (2007-2012). The model outputs were in good agreement with the peak of nitrite downstream the WWTP as well as its slow decrease towards the estuary. Nitrite persistence between the WWTP and the estuary was mostly explained by similar production and consumption rates of nitrite in both water and sediment layers. The sediment layer constituted a significant source of nitrite, especially during high river discharges (0.1-0.4 mgN h(-1) m(-2)). This points out how essential it is to represent the benthic layer in river water quality models, since it can constitute a source of nitrite to the water-column. As a consequence of anthropogenic emissions and in-river processes, nitrite fluxes to the estuary were significant and varied from 4.1 to 5.5 TN d(-1) in low and high water discharge conditions, respectively, over the 2007-2012 period. This study provides a methodology that can be applied to any anthropized river to realistically parametrize autochthonous and WWTP-related nitrifier communities and simulate nitrite dynamics. Based on simulation analysis, it is shown that high spatio-temporal resolution hydro-ecological models are efficient to 1) estimate water quality criteria and 2) forecast the effect of future management strategies. Process-based simulations constitute essential tools to complete our understanding of nutrient cycling, and to decrease monitoring costs in the context of water quality and eutrophication management in river ecosystems.


Alteration of the irisin-brain-derived neurotrophic factor axis contributes to disturbance of mood in COPD patients.
COPD is accompanied by limited physical activity, worse quality of life, and increased prevalence of depression. A possible link between COPD and depression may be irisin, a myokine, expression of which in the skeletal muscle and brain positively correlates with physical activity. Irisin enhances the synthesis of brain-derived neurotrophic factor (BDNF), a neurotrophin involved in reward-related processes. Thus, we hypothesized that mood disturbances accompanying COPD are reflected by the changes in the irisin-BDNF axis. Case history, routine laboratory parameters, serum irisin and BDNF levels, pulmonary function, and disease-specific quality of life, measured by St George's Respiratory Questionnaire (SGRQ), were determined in a cohort of COPD patients (n=74). Simple and then multiple linear regression were used to evaluate the data. We found that mood disturbances are associated with lower serum irisin levels (SGRQ's Impacts score and reciprocal of irisin showed a strong positive association; β: 419.97; 95% confidence interval [CI]: 204.31, 635.63; P<0.001). This association was even stronger among patients in the lower 50% of BDNF levels (β: 434.11; 95% CI: 166.17, 702.05; P=0.002), while it became weaker for patients in the higher 50% of BDNF concentrations (β: 373.49; 95% CI: -74.91, 821.88; P=0.1). These results suggest that irisin exerts beneficial effect on mood in COPD patients, possibly by inducing the expression of BDNF in brain areas associated with reward-related processes involved in by depression. Future interventional studies targeting the irisin-BDNF axis (eg, endurance training) are needed to further support this notion.


Planning for emergency medical services in Boston.
The results of a survey of 10,200 visits to 11 Boston hospital emergency rooms during a 9-day period in March 1972 are presented. The survey was designed to provide data on emergency room use to permit more informed planning by public agencies concerned with improving areawide emergency medical services. The 11 institutions surveyed provided virtually all of the emergency medical services in the city of Boston. A majority are teaching hospitals affiliated with one or more of the three medical schools in the area. Of the 11 hospitals, 3 accounted for 60% of all emergency room visits. Survey data were extracted from emergency room log sheets and hospital medical records of individual patients. Information collected included the residence pattern of patients within the geographic area, the patient mix by degree of urgency based on presenting complaints, mode of transportation to the hospital, and age and sex of the patients. Only 15 percent of the 10,200 visits were true emergencies. Fifty-seven percent were classified as urgant and 28% nonurgent. The mix among the 11 hospitals ranged from 7 to 22 percent in the emergency category, and 11 to 61 percent in the nonurgent classification. Trauma accounted for 19 percent of all admissions, with 3 percent attributed to fractures and 4 percent to head injuries. Fifty-six percent of the emergency cases required the services of an internist or pediatrician, 38 percent a surgeon, and 1 percent an obstetrician. The highest utilization rate--27 per 1,000 population--was recorded for the under 5 age group. Although the 65 and older age group had the lowest utilization rate of 6 per 1,000, this group had the highest rate of visits classified as emergencies. Children under 5 accounted for the highest proportion of nonurgent visits. The survey revealed that 30 percent of all hospital admissions were from the emergency room. One in four emergency patients lived outside the city of Boston. A neighborhood health center and a hospital general practice unit reduced hospital emergency room workloads appreciably, even when they were open only during daytime hours. Eighty-eight percent of all patients arranged for their own transportation, usually by private automobile. Of those arriving by ambulance, only 35 percent were classified as emergencies. The survey data reinforce the conclusion that major planning efforts should be concentrated on the management of the nonemergency patient. The data also emphasize the need for a single agency to be responsible for overall planning for emergency medical services on an area wide basis.


[To the differential diagnosis of arthritis of pseudotuberculous and chlamydial etiology].
to define main differential diagnostic criteria for arthritides of chlamydial and pseudotuberculous etiology and to improve patient examination tactics. Forty-six patients with pseudotuberculosis and 41 patients with chronic urogenital chlamydial infection with articular involvement were examined. A bacteriological method of polymerase chain reaction (PCR), agglutination test, enzyme immunoassay (EIA) (IgA, IgG, IgM), indirect hemagglutination (IHA) test were used to diagnose pseudotuberculosis. Diagnostic techniques for chlamydiasis involved cultural, direct immunofluorescence (DIF), real-time PCR, and EIA (IgM, IgG, IgA). Patients with pseudotuberculosis developed polyarthritis and oligoarthritis in 56 and 39%, respectively. The development of arthritides was accompanied by fever in 89%, exanthema in 57%, gastrointestinal lesion in 56%, hepatomegalia in 78%. The pseudotuberculous etiology of the disease was confirmed by the agglutination test in 71% of the patients and by IHA in 7%. EIA revealed IgG in 78% of the patients, IgA in 11%, and IgM in 29%. PCR of synovial fluid (SF), synovial shell, and other biological substrates revealed Yersinia pseudotuberculosis DNA in 43%. Chlamydiasis and polyarthritis developed in 71 and 19%, respectively. The diagnosis of chlamydiasis was verified by EIA detection of IgG and IgA in 76 and 27% of cases, respectively. DIF, PCR, and culture studies of urethral scrapes found Chlamydia in 9, 32, and 29% of cases, respectively. Examination of SF and synovial shells revealed Chlamydia trachomatis in 24% of the patients and culture studies detected the pathogen in 21%. Asymmetrical polyarthritides mainly involving the knee joints are the most common arthritides of pseudotuberculous etiology. EIA detection of serum IgG and IgA and PCR study of SF are optimal diagnostic tools. Artritides of chlamydial etiology are asymmetrical oligoarthritides predominantly involving the knee and ankle joints. Examination of urethral and cervical canal scrapes, SF by culture and PCR studies and that of serum IgA and IgG by EIA are optimal diagnostic tests.


Adaptive Processes in Hearing.
Our auditory environment is constantly changing and evolving over time, requiring us to rapidly adapt to a complex dynamic sensory input. This adaptive ability of our auditory system can be observed at different levels, from individual cell responses to complex neural mechanisms and behavior, and is essential to achieve successful speech communication, correct orientation in our full environment, and eventually survival. These adaptive processes may differ in individuals with hearing loss, whose auditory system may cope via "readapting" itself over a longer time scale to the changes in sensory input induced by hearing impairment and the compensation provided by hearing devices. These devices themselves are now able to adapt to the listener's individual environment, attentional state, and behavior. These topics related to auditory adaptation, in the broad sense of the term, were central to the 6th International Symposium on Auditory and Audiological Research held in Nyborg, Denmark, in August 2017. The symposium addressed adaptive processes in hearing from different angles, together with a wide variety of other auditory and audiological topics. The papers in this special issue result from some of the contributions presented at the symposium.


Cytoplasmic CCR7 (CCR7c) Immunoexpression Is Associated with Tumor Invasion in Gastric Cancer.
Background : This study investigates the CCR7 chemokine receptor's prognostic value in gastric cancer and its relationship to metastasis. Materials and Methods : Normal and adjacent tumor cells in 70 patients with gastric cancer were evaluated for CCR7 expression using immunohistochemical staining. The prognostic values of high and low levels of expression of CCR7 were also evaluated by multivariate and univariate analyses. Results : Analysis indicated high expression of CCR7 in 52.9% of tumor tissue. Moreover, high expression of CCR7 was significantly related to metastasis of lymph nodes (p = 0.00). In addition, high expression of CCR7 had a positive correlation to the disease stage (p = 0.00), age of ≥50 years (p = 0.019), male gender (p = 0.024), vascular involvement (p = 0.009), histology of tumor adenocarcinoma (p = 0.00), and poor tumor differentiation (p = 0.00). However, the high expression of the CCR7 marker was not related to the tumor size. Conclusion : Based on our results, CCR7 expression in gastric cancer can be considered a clinical prognostic indicator in patients with gastric cancer.


Correction of parallel transmission using concurrent RF and gradient field monitoring.
The accuracy and precision of the parallel RF excitations are highly dependent on the spatial and temporal fidelity of the magnetic fields involved in spin excitation. The consistency between the nominal and effective fields is typically limited by the imperfections of the employed hardware existing both in the gradient system and the RF chain. In this work, we experimentally presented highly improved spatially tailored parallel excitations by turning the native hardware accuracy challenge into a measurement and control problem using an advanced field camera technology to fully correct parallel RF transmission experiment. An array of NMR field probes is used to measure the multiple channel RF pulses and gradient waveforms recording the high power RF pulses simultaneously with low frequency gradient fields on equal time basis. The recorded waveforms were integrated in RF pulse design for gradient trajectory correction, time imperfection compensation and introduction of iterative RF pre-emphasis. Superior excitation accuracy was achieved. Two major applications were presented at 7 Tesla including multi-dimensional tailored RF pulses for spatially selective excitation and slice-selective spoke pulses for [Formula: see text] mitigation. Comprehensive field monitoring is a highly effective means of correcting for the field deviations during parallel transmit pulse design.


Mutational analysis of Glu-327 of Na(+)-K(+)-ATPase reveals stimulation of 86Rb+ uptake by external K+.
A competition assay of 86Rb+ uptake in HeLa cells transfected with ouabain-resistant Na(+)-K(+)-ATPase mutants revealed a stimulation of 86Rb+ uptake at low external concentrations (1 mM) of competitor (K+). Of the models that were tested, those that require that two K+ be bound before transport occurs gave the worst fits. Random and ordered binding schemes described the data equally well. General models in which both binding and transport were allowed to be cooperative yielded parameter errors larger than the parameters themselves and could not be utilized. Models that assumed noncooperative transport always showed positive cooperativity in binding. E327Q and E327L mutated forms of rat alpha 2 had lower apparent affinities for the first K+ bound than did wild-type rat alpha 2 modified to be ouabain resistant. The mutations did not affect the apparent affinity of the second K+ bound. Models that assumed noncooperativity in binding always showed positively cooperative transport, i.e., enzymes with two K+ bound had a higher flux than those with one K+ bound. Increases in external Na+ decreased the apparent affinity for K+ for all models and decreased the ratio of the apparent influx rate constants for E327L.


System performance evaluation of refractive surgical lasers: a mathematical approach.
A study was conducted for the purpose of improving the designs of the next generation of refractive surgical laser systems. Two common refractive laser systems, variable-spot scanning (type A) and small-spot scanning (type B), are discussed by identifying sources of error that could adversely affect the capability of these lasers to accurately produce complex, customized wavefront guided ablations. A mathematical model was used to construct a laser simulator that models the two common laser systems in terms of the root-mean-square error. Error sources from ablation profile fitting, ablation registration, eye tracking, and the laser delivery system are compared, and the relative contribution of each to the overall system error is analyzed. This system-level analysis can be helpful to the improvement of both laser systems.


FGF19 action in the brain induces insulin-independent glucose lowering.
Insulin-independent glucose disposal (referred to as glucose effectiveness [GE]) is crucial for glucose homeostasis and, until recently, was thought to be invariable. However, GE is reduced in type 2 diabetes and markedly decreased in leptin-deficient ob/ob mice. Strategies aimed at increasing GE should therefore be capable of improving glucose tolerance in these animals. The gut-derived hormone FGF19 has previously been shown to exert potent antidiabetic effects in ob/ob mice. In ob/ob mice, we found that systemic FGF19 administration improved glucose tolerance through its action in the brain and that a single, low-dose i.c.v. injection of FGF19 dramatically improved glucose intolerance within 2 hours. Minimal model analysis of glucose and insulin data obtained during a frequently sampled i.v. glucose tolerance test showed that the antidiabetic effect of i.c.v. FGF19 was solely due to increased GE and not to changes of either insulin secretion or insulin sensitivity. The mechanism underlying this effect appears to involve increased metabolism of glucose to lactate. Together, these findings implicate the brain in the antidiabetic action of systemic FGF19 and establish the brain's capacity to rapidly, potently, and selectively increase insulin-independent glucose disposal.


A randomized, double-blind trial of filgrastim (granulocyte colony-stimulating factor) versus placebo following allogeneic blood stem cell transplantation.
Blood stem cell transplantation (BSCT) results in rapid hematopoietic recovery in both the allogeneic and autologous transplant settings. Because of the large numbers of progenitor cells in mobilized blood, the administration of growth factors after transplantation may not provide further acceleration of hematopoietic recovery. A randomized, double-blind, placebo-controlled study was performed to determine the effects of filgrastim (granulocyte colony-stimulating factor; G-CSF) administration on hematopoietic recovery after allogeneic BSCT. Fifty-four patients with hematologic malignancies undergoing a related, HLA-matched allogeneic BSCT were randomly assigned to receive daily filgrastim at 10 microg/kg or placebo starting on the day of transplantation. A minimum of 3 x 10(6) CD34(+) cells/kg in the allograft was required for transplantation. All patients received a standard preparative regimen and a standard regimen for the prevention of graft-versus-host disease (GVHD). The median time to achieve an absolute neutrophil count greater than 0.5 x 10(9)/L was 11 days (range, 9-20 days) for patients who received filgrastim compared with 15 days (range, 10-22 days) for patients who received placebo (P =.0082). The median time to achieve a platelet count greater than 20 x 10(9)/L was 13 days (range, 8-35 days) for patients who received filgrastim compared with 15.5 days (range, 8-42 days) for patients who received placebo (P =.79). There were no significant differences for red blood cell transfusion independence, the incidence of acute GVHD, or 100-day mortality between the groups. The administration of filgrastim appears to be a safe and effective supportive-care measure following allogeneic BSCT.


Taspase1-dependent TFIIA cleavage coordinates head morphogenesis by limiting Cdkn2a locus transcription.
Head morphogenesis requires complex signal relays to enable precisely coordinated proliferation, migration, and patterning. Here, we demonstrate that, during mouse head formation, taspase1-mediated (TASP1-mediated) cleavage of the general transcription factor TFIIA ensures proper coordination of rapid cell proliferation and morphogenesis by maintaining limited transcription of the negative cell cycle regulators p16Ink4a and p19Arf from the Cdkn2a locus. In mice, loss of TASP1 function led to catastrophic craniofacial malformations that were associated with inadequate cell proliferation. Compound deficiency of Cdkn2a, especially p16Ink4a deficiency, markedly reduced the craniofacial anomalies of TASP1-deficent mice. Furthermore, evaluation of mice expressing noncleavable TASP1 targets revealed that TFIIA is the principal TASP1 substrate that orchestrates craniofacial morphogenesis. ChIP analyses determined that noncleaved TFIIA accumulates at the p16Ink4a and p19Arf promoters to drive transcription of these negative regulators. In summary, our study elucidates a regulatory circuit comprising proteolysis, transcription, and proliferation that is pivotal for construction of the mammalian head.


Prediction of long-term disability in multiple sclerosis.
Little is known about the predictive value of neurophysiological measures for the long-term course of multiple sclerosis (MS). To prospectively investigate whether combined visual (VEP) and motor evoked potentials (MEP) allow prediction of disability over 14 years. A total of 30 patients with relapsing-remitting and secondary progressive MS were prospectively investigated with VEPs, MEPs and the Expanded Disability Status Scale (EDSS) at entry (T0) and after 6, 12 and 24 months, and with cranial MRI scans at entry (T2-weighted and gadolinium-enhanced T1-weighted images). EDSS was again assessed at year 14 (T4). The association between evoked potential (EP), magnetic resonance (MR) data and EDSS was measured using Spearman's rank correlation. Multivariable linear regression was performed to predict EDSS(T4) as a function of z-transformed EP-latencies(T0). The model was validated using a jack-knife procedure and the potential for improving it by inclusion of additional baseline variables was examined. EDSS values(T4) correlated with the sum of z-transformed EP-latencies(T0) (rho = 0.68, p < 0.0001), but not with MR-parameters(T0). EDSS(T4) as predicted by the formula EDSS(T4) = 4.194 + 0.088 * z-score P100(T0) + 0.071 * z-score CMCT(UE, T0) correlated with the observed values (rho = 0.69, p < 0.0001). Combined EPs allow prediction of long-term disability in small groups of patients with MS. This may have implications for the choice of monitoring methods in clinical trials and for daily practice decisions.


MR imaging assessment of myocardial edema with T2 mapping.
Cardiac magnetic resonance (CMR) provides a high signal-to-noise ratio, high spatial and temporal resolutions, as well as a delayed-enhancement sequence and is therefore considered a reference technique in the field of cardiac imaging. However, currently available sequences are not adequate to assess some pathologic conditions, such as myocardial edema. T2 mapping sequences generate parametric images that are based on the transverse relaxation time (T2) for each voxel. In case of edema, the T2 relaxation time is longer. This review summarizes current knowledge on CMR T2 mapping for assessing myocardial edema.


Additional chromosome abnormalities, BCR-ABL tyrosine kinase domain mutations and clinical outcome in Hungarian tyrosine kinase inhibitor-resistant chronic myelogenous leukemia patients.
Additional chromosome abnormalities (ACAs), mutations of the BCR-ABL tyrosine kinase domain (TKD) and BCR-ABL splice variants may cause resistance to first- and second-generation tyrosine kinase inhibitors (TKIs) in chronic myelogenous leukemia (CML) and Philadelphia-positive (Ph+) acute lymphoid leukemia (ALL). Karyotyping and BCR-ABL TKD mutation screening were performed in 71 imatinib-resistant CML patients and 6 Ph+ ALL patients. A total of 56 out of these 77 patients received second-generation TKI. ACAs were present in 30 of 65 imatinib-resistant patients (46%). In 27 of 74 imatinib-resistant patients (36%), 15 different BCR-ABL TKD mutations were detected. Mutations were found in 25% of chronic-phase patients (12/47), 33% of accelerated-phase patients (5/15), 71% of blast crisis CML patients (5/7) and 100% of ALL patients. In nilotinib-resistant patients, Y253H, T315I and F359I/V mutations were detected; in dasatinib-resistant patients, L248M, E279K and T315I mutations were detected. T315I was found more frequently in patients on dasatinib than on imatinib therapy. The presence of ACAs predicted shorter survival during first- and second-generation TKI therapy, while TKD mutations only influenced survival during second-generation TKI therapy. For patients with TKI resistance, mutation and ACA screening may play a role in identifying patients with poorer prognosis.


Post-surgical lipophagic panniculitis: a specific model of traumatic panniculitis and new histopathological findings.
Aim of this work was to define the histopathological features of post-surgical panniculitis. Dermal and hypodermal changes will be analyzed in detail, to understand the cascade of events that characterize the tissue response to surgical trauma. Cutaneous re-excision specimens of cases of basal cell carcinoma, squamous cell carcinoma, and melanoma consecutively seen from January 1, 2011 to June 30, 2011 at the Department of Dermatology, University of Pavia, were included in this study. Only the cases in which the first surgical procedure included the subcutaneous fat, were considered. In addition, the time elapsed from the first surgical procedure and the re-excision had to be included in a period of time from one to three months. All the specimens were stained with hematoxylin and eosin. Thirty cutaneous re-excision specimens were studied. Histopathologic examination revealed changes of epidermis, ranging from slight atrophy to moderate hyperplasia. In two cases focal ulceration was seen, with transfollicular elimination of foreign body material. The main dermal changes observed were the: 1) scar with well defined vertical orientation along the dermal suture line; 2) rounded cicatricial areas with radial branching septa of scarring tissue; 3) foreign body granuloma formation; 4) alignment of hystiocytes at the dermo-hypodermal border; 5) traumatic neuromas. The subcutaneous fat changes included: 1) lobular panniculitis with consistent presence of foam cells; 2) striking anisocytosis with pseudocystic degeneration and necrosis of adipocytes; 3) eritrocyte extravasation, mainly at the dermo-hypodermal border; 4) deep seated phlebitis. Post-surgical panniculitis is a lobular foam cell panniculitis characterized by simultaneous dermal and hypodermal changes, expression of the multi-faceted tissue response to a surgical trauma. This type of peculiar lipophagic response puts post-surgical panniculitis into the wider chapter of lipophage tissue response seen in atherosclerosis, glomerulosclerosis and some infectious models such as Mycobacterium tuberculosis and Chlamydia pneumoniae infections. Furthermore it may be seen as a reliable and convenient model for laboratory investigation on foam cell tissue response.


First report of natural infection of least weasel (Mustela nivalis Linnaeus, 1776) with Leishmania major in Tunisia.
Zoonotic cutaneous leishmaniasis, caused by Leishmania major (L. major), is endemic in Tunisia. Several rodents have been identified as reservoir hosts of parasites. This study reports, for the first time, the natural infection with L. major zymodeme MON-25 in a specimen of least weasel: Mustela nivalis Linnaeus, 1776 (M. nivalis) collected in Sidi Bouzid. This finding justifies further research on larger samples of this animal to verify its role as a potential reservoir host for cutaneous leishmaniasis in Tunisia.


Psychosocial correlates of dietary behaviour in type 2 diabetic women, using a behaviour change theory.
The study evaluated the efficacy of the Theory of Reasoned Action (TRA), along with self-efficacy to predict dietary behaviour in a group of Iranian women with type 2 diabetes. A sample of 352 diabetic women referred to Khoy Diabetes Clinic, Iran, were selected and given a self-administered survey to assess eating behaviour, using the extended TRA constructs. Bivariate correlations and Enter regression analyses of the extended TRA model were performed with SPSS software. Overall, the proposed model explained 31.6% of variance of behavioural intention and 21.5% of variance of dietary behaviour. Among the model constructs, self-efficacy was the strongest predictor of intentions and dietary practice. In addition to the model variables, visit intervals of patients and source of obtaining information about diabetes from sociodemographic factors were also associated with dietary behaviours of the diabetics. This research has highlighted the relative importance of the extended TRA constructs upon behavioural intention and subsequent behaviour. Therefore, use of the present research model in designing educational interventions to increase adherence to dietary behaviours among diabetic patients was recommended and emphasized.


Marrying the radical, the conventional, and the mystical: Mathematics, gender and religion in the lives of William Kingdon and Lucy Lane Clifford.
The avowed atheist, evolutionary naturalist and mathematician William Kingdon Clifford is often remembered for his essay, "The Ethics of Belief," in which he opposed organized religion in any form. As a mathematician, Clifford was an early advocate of non-Euclidean geometry in England. Combining William Rowan Hamilton's work on quaternions with Hermann Grassmann's theory of linear extension, he invented an original system of geometric algebra. Breaking with conservative traditionalism in his philosophical and mathematical work, Clifford's marriage to the children's writer, novelist, and dramatist Lucy Lane was a relatively conventional, if brief, Victorian marriage. After his untimely death from consumption in 1879, Lucy outlived her husband by fifty years. Raising their two daughters and supporting herself after his passing, Lucy refashioned Clifford's posthumous reputation to temper his philosophical radicalism. Her collaboration with Clifford's publisher and editor reveal Lucy's concern that Clifford not be remembered as someone ruled by passion in his mathematical work. Her efforts to expunge writings suggestive of William's weakness, excitability, or inconstancy from the public record demonstrates her desire to craft an image of her husband in alignment with gendered expectations of masculinity. This paper argues that Lucy fashioning of William's memory conformed, rather than departed from, normative parameters of gender as defined by Victorian society.


Clinical manifestations, therapy and outcome of pandemic influenza A (H1N1) 2009 in hospitalized patients.
Increasing number of epidemiological and clinical studies to date showed that the pandemic influenza A (H1N1) 2009, by its characteristics, significantly differs from infection caused by seasonal influenza. Therefore, the information about clinical spectrum of manifestations, risk factors for severe form of the disease, treatment and outcome in patients with novel flu are still collected. METHODS; A total of 98 patients (mean age 32 +/- 15 years, range 14-88 years) with the signs and symptoms of novel influenza were treated in the Clinic for Infectious and Tropical Diseases, Military Medical Academy. There were 74 (75.5%) patients with suspected influenza A (H1N1) 2009, 10 (10.2%) with the likelihood and 14 (14.3%) with the confirmed influenza. In all the patients we registered the basic demographic data, risk factors for severe disease, symptoms and signs of influenza, laboratory tests and chest radiography. We analyzed antiviral therapy use and disease outcome (survived, died). The average time from the beginning of influenza A (H1N1) to the admission in hospital was 3 days (0-16 days) and from the moment of hospitalization to the Intensive Care Unit (ICU) admission was 2 days (0-5 days). There were 49 (50.0) patients, 20-29 years of age and 5 (5.1) patients older than 65. A total of 21 (21.4%) patients were with underlying disease, 18 (18.4%) were obese, 19 (19.4) were cigarette smokers. All of the patients had fever, 81 (82.6%) cough, while dyspnea and diarrhea were registered in 4 of the patients. In more than 75% of the patients laboratory tests were within normal limits. The real-time polymerase chain reaction (PCR) test for identification of influenza A (H1N1) 2009 was positive in 14 (77.8%), while pneumonia was verified in 30 (30.7%) of the patients. Six (6.1%) patients, mean age of 45 +/- 14 years (31-59 years) were admitted to the ICU, of whom five (5.1%) had Adult Respiratory Distress Syndrome (ARDS). Risk factors were registered more frequently in the patients with acute respiratory failure (14.2% vs 4.9%, p < 0.05). A total of 67 (68.4%) patients received oseltamivir, 89 (90.1%) was applied to antibiotics and 64 (65.3%) were treated with a combined therapy. Antiviral therapy was applied to 43 (43.3%) patients in the first 48 hours from the onset of the disease, of whom only one (3.4) developed ARDS. Fatal outcome was noted in 2.0% of the patients (2 of 98 patients) and in 33.3% of the patients treated in the ICU. Novel influenza A (H1N1) is most commonly manifested as a mild acute respiratory disease, which usually affects young healthy adults. A small number of the patients develop severe illness with acute respiratory failure and death. Patients seem to have benefit from antiviral therapy especially in first 48 hours.


Clinical correlates of olfactory hallucinations in schizophrenia.
OBJECTIVES. Olfactory hallucinations (OHs) are underrepresented in conventional clinical instruments, infrequently researched, and poorly understood. To advance understanding of OHs, we examined their past-month prevalence and co-occurring symptoms in two datasets. DESIGN. One dataset comprised categorical codes and was examined using homogeneity analysis and logistic regression; the other dataset comprised numeric ratings and was examined using principal components analyses and linear regression. METHOD. The two datasets included: (1) 962 cases with Present State Examination - 9th Edition (PSE-9), codes (recoded present/absent) from the World Health Organization 10 Country (WHO-10) Study and (2) 265 cases with ratings on Scales for Assessing Positive/Negative Symptoms of Schizophrenia (SAPS/SANS). Two PSE-9 items (external- and self-smells) were recoded into a single OH item to examine consistency with the SAPS/SANS dataset, which contained a single OH item. RESULTS. Prevalence of OHs and hallucinations in other modalities differed according to the WHO-10 international centre. Across centres, OHs were present in 13% of the WHO-10 dataset, similar to the 17% prevalence rate in the SAPS/SANS dataset. Referential/control delusions and other hallucinations (particularly, somatic/tactile/gustatory hallucinations) were significant independent correlates of OHs in both datasets. OHs also co-occurred with social anxiety and depression in the WHO-10 dataset, with self-smells being particularly associated with self-depreciation. CONCLUSIONS. Sociocultural factors may modulate the self-reporting and/or detection of OHs and hallucinations in other modalities. Referential/control delusions promote the generation and/or maintenance of OHs independent of factors shared with other hallucinations. OHs and hallucinations of taste, touch, and bodily sensation frequently co-occur. Self-smells warrant sensitive probing.


The Caenorhabditis elegans nuclear receptor gene nhr-25 regulates epidermal cell development.
The development of the epidermis of Caenorhabditis elegans involves cell fusion, migration, and differentiation events. To understand the mechanisms underlying these processes, we characterized the roles of NHR-25, a member of the nuclear receptor family of transcription factors. The NHR-25 homologs Ftz-F1 in Drosophila and SF-1 in mammals are involved in various biological processes, including regulation of patterning during development, reproduction, metabolism, metamorphosis, and homeostasis. Impairment of nhr-25 activity leads to severe phenotypes in embryos and many postembryonic tissues. Further analysis has indicated that nhr-25 activity is required for the proper development, including cell-cell fusion, of several epidermal cell types, such as the epidermal syncytial, seam, and Pn.p cells. Our results also suggest that nhr-25 is likely to regulate cell-cell junctions and/or fusion. In a subset of Pn.p cells, called vulval precursor cells, nhr-25 acts collaboratively with the lin-39 Hox gene in regulating vulval cell differentiation. Additionally, our data suggest that nhr-25 may also function with another Hox gene, nob-1, during embryogenesis. Overall, our results indicate that nhr-25 plays an integral role in regulating cellular processes of epidermal cells.


The effect of photobiomodulation on hearing loss: A systematic review.
To assess outcomes associated with photobiomodulation therapy (PBMT) for hearing loss in human and animal studies. Systematic review and narrative synthesis in accordance with PRISMA guidelines. Data bases searched: MEDLINE, EMBASE, CENTRAL, ClinicalTrials.gov and Web of Science. No limits were placed on language or year of publication. Review conducted in accordance with the PRISMA 2020 statement. All human and animal subjects treated with PBMT for hearing loss. Pre- and post-PBMT audio metric outcomes. Searches identified 122 abstracts and 49 full text articles. Of these, 17 studies met the inclusion criteria, reporting outcomes in 327 animals (11 studies), 30 humans (1 study), and 40 animal specimens (5 studies). PBMT parameters included 6 different wavelengths: 908 nm (1 study), 810 nm (1 study), 532 & 635 nm (1 study), 830 nm (3 studies), 808 nm (11 studies). The duration ranged from 4 to 60 minutes in a session, and the follow-up ranged from 5-28 days. Outcomes improved significantly when wavelengths within the range of 800-830 nm were used, and with greater duration of PBMT exposure. Included studies predominantly consisted of non-randomized controlled trials (10 studies). Hearing outcomes following PBMT appear to be superior to no PBMT for subjects with hearing loss, although higher level evidence is required to verify this. PBMT enables concentrated, focused delivery of light therapy to the inner ear through a non-invasive manner with minimal side effects. As a result of heterogeneity in reporting PBMT parameters and outcomes across the included studies, direct comparison is challenging.


Blockade of GABA(A) receptors in the ventromedial hypothalamus further stimulates glucagon and sympathoadrenal but not the hypothalamo-pituitary-adrenal response to hypoglycemia.
Hypoglycemia provokes a multifaceted counterregulatory response involving the sympathoadrenal system, stimulation of glucagon secretion, and the hypothalamo-pituitary-adrenal axis that is commonly impaired in diabetes. We examined whether modulation of inhibitory input from gamma-aminobutyric acid (GABA) in the ventromedial hypothalamus (VMH), a major glucose-sensing region within the brain, plays a role in affecting counterregulatory responses to hypoglycemia. Normal Sprague-Dawley rats had carotid artery and jugular vein catheters chronically implanted, as well as bilateral steel microinjection guide cannulas inserted down to the level of the VMH. Seven to 10 days following surgery, the rats were microinjected with artificial extracellular fluid, the GABA(A) receptor agonist muscimol (1 nmol/side), or the GABA(A) receptor antagonist bicuculline methiodide (12.5 pmol/side) before being subjected to a hyperinsulinemic-hypoglycemic (2.5 mmol/l) glucose clamp for 90 min. Following VMH administration of bicuculline methiodide, glucose infusion rates were significantly suppressed, whereas muscimol raised glucose infusion rates significantly compared with controls. Glucagon and epinephrine responses were elevated with the antagonist and suppressed with the agonist compared with controls. Corticosterone responses, however, were unaffected by either administration of the agonist or antagonist into the VMH. These data demonstrate that modulation of the GABAergic system in the VMH alters both glucagon and sympathoadrenal, but not corticosterone, responses to hypoglycemia. Our findings are consistent with the hypothesis that GABAergic inhibitory tone within the VMH can modulate glucose counterregulatory responses.


Identification of XMRV infection-associated microRNAs in four cell types in culture.
XMRV is a gammaretrovirus that was thought to be associated with prostate cancer (PC) and chronic fatigue syndrome (CFS) in humans until recently. The virus is culturable in various cells of human origin like the lymphocytes, NK cells, neuronal cells, and prostate cell lines. MicroRNAs (miRNA), which regulate gene expression, were so far not identified in cells infected with XMRV in culture. Two prostate cell lines (LNCaP and DU145) and two primary cells, Peripheral Blood Lymphocytes [PBL] and Monocyte-derived Macrophages [MDM] were infected with XMRV. Total mRNA was extracted from mock- and virus-infected cells at 6, 24 and 48 hours post infection and evaluated for microRNA profile in a microarray. MicroRNA expression profiles of XMRV-infected continuous prostate cancer cell lines differ from that of virus-infected primary cells (PBL and MDMs). miR-193a-3p and miRPlus-E1245 observed to be specific to XMRV infection in all 4 cell types. While miR-193a-3p levels were down regulated miRPlus-E1245 on the other hand exhibited varied expression profile between the 4 cell types. The present study clearly demonstrates that cellular microRNAs are expressed during XMRV infection of human cells and this is the first report demonstrating the regulation of miR193a-3p and miRPlus-E1245 during XMRV infection in four different human cell types.


Hyperspectral imaging to predict the effect of cyclophosphamide in primary membranous nephropathy.
Presently, there is a lack of accurate predictors of the efficacy of primary membranous nephropathy. The aim of this study is to explore the application value of hyperspectral imaging in predicting the efficacy of cyclophosphamide treatment in primary membranous nephropathy. A total of 30 patients with primary membranous nephropathy who were treated with glucocorticoid combined with cyclophosphamide were collected. Hematoxylin-eosin stained renal pathological images were acquired by hyperspectral imaging system at the spectral range of 400-1000 nm. The remission group data set contained 23 samples, while the non-remission group data set contained 28 samples. A one-dimensional convolutional neural network model was established to train and test the hyperspectral data, and the performance of the model was evaluated. From the spectral curve, the spectral difference between the remission group and the non-remission group was obvious between 525 and 700 nm. The spectral data in this band were analyzed by one-dimensional convolutional neural network, and the confusion matrix showed that the remission group and the non-remission group were successfully classified. The precision and recall were 0.89 and 0.81 for the non-response group and 0.83 and 0.90 for the response group, respectively, with an F1 score of 0.85 in both groups. The area under the AUC curve of the classification model reached 0.857. In this study, a one-dimensional convolutional neural network model was used to analyze the hyperspectral images of renal pathology of PMN patients, and the patients in remission group and non-remission group were successfully classified after glucocorticoid combined with cyclophosphamide treatment.


In vivo photodynamic activity of hypericin in transitional cell carcinoma bladder tumors.
In a recent clinical study, we showed that hypericin accumulates selectively in urothelial lesions of the bladder following intravesical administration of the compound in patients. This observation infers that hypericin, a potent photosensitizer, could be used as a selective photodynamic therapy (PDT) tool against superficial bladder cancer. In the present study we investigated the in vivo PDT activity of hypericin in transition cell carcinoma (TCC) tumors of the bladder. Both the distribution and tumor PDT response were carried out using subcutaneous heterotopic AY-27 TCC tumors in syngeneic rats. For both PDT and distribution studies, hypericin (1 or 5 mg/kg) was injected intravenously 0.5, 6 or 24 h before PDT or distribution evaluation. The data show that hypericin is a potent photosensitizer in the treatment of TCC tumors in vivo and that the interval between drug administration and photo-irradiation has a dramatic effect on the PDT outcome. Using a 0.5 h interval between drug administration and photo-irradiation the tumor regrowth study indicated that no tumor mass could me measured 9-10 days after PDT. On the contrary, lengthening the time interval between drug administration and photo-irradiation resulted in a gradual loss of PDT efficiency in these tumors. For instance, while the 6 h drug interval protocol produced a moderate PDT activity in which the tumor sizes decreased to about 50% of their original sizes 11-16 days after photo-irradiation, the 24 h interval protocol was even less effective. The distribution data indicate that the PDT efficiency of hypericin in TCC tumors corresponded to the plasma concentrations rather than to the over all concentrations in the tumor. It is therefore conceivable that the mechanism of PDT efficacy of hypericin in TCC tumors is through indirect (vascular effects) rather than through direct effects (cellular destruction) of hypericin in these tumors. In conclusion, our data indicate that hypericin is a potent photosensitizer against AY-27 TCC tumors and that the PDT efficacy of hypericin is largely determined by photosensitizer distribution in the tumor at the time of photo-irradiation.


Molecular geometries of the heme axial ligands from the triheme cytochrome PpcF from Geobacter metallireducens reveal a conserved heme core architecture.
Electroactive Geobacter bacteria can perform extracellular electron transfer and present a wide metabolic versatility. These bacteria reduce organic, toxic and radioactive compounds, and produce electric current while interacting with electrodes, making them interesting targets for numerous biotechnological applications. Their global electrochemical responses rely on an efficient interface between the inside and the cell's exterior, which is driven by the highly abundant periplasmic triheme PpcA-family cytochromes. The functional features of these cytochromes have been studied in G. sulfurreducens and G. metallireducens, and although they share a high degree of structural homology and sequence identity, their properties are quite distinct. In this work, the heme axial ligand geometries and the magnetic properties of PpcF from G. metallireducens were determined. The data obtained constitute important constraints for the determination of its solution structure in the oxidized state and indicate that the (i) heme core architecture; (ii) axial ligands geometries and (iii) magnetic properties of the cytochrome are conserved compared to the other members of the PpcA-families. Furthermore, the results also indicate that the heme arrangement is crucial to maintain an intrinsic regulation of the protein's redox properties and hence its electron transfer efficiency and functionality.


The intrinsic memorability of face photographs.
The faces we encounter throughout our lives make different impressions on us: Some are remembered at first glance, while others are forgotten. Previous work has found that the distinctiveness of a face influences its memorability--the degree to which face images are remembered or forgotten. Here, we generalize the concept of face memorability in a large-scale memory study. First, we find that memorability is an intrinsic feature of a face photograph--across observers some faces are consistently more remembered or forgotten than others--indicating that memorability can be used for measuring, predicting, and manipulating subsequent memories. Second, we determine the role that 20 personality, social, and memory-related traits play in face memorability. Whereas we find that certain traits (such as kindness, atypicality, and trustworthiness) contribute to face memorability, they do not suffice to explain the variance in memorability scores, even when accounting for noise and differences in subjective experience. This suggests that memorability itself is a consistent, singular measure of a face that cannot be reduced to a simple combination of personality and social facial attributes. We outline modern neuroscience questions that can be explored through the lens of memorability.


Discerning the mechanism of action of HtrA4: a serine protease implicated in the cell death pathway.
High-temperature requirement protease A4 (HtrA4) is a secretary serine protease whose expression is up-regulated in pre-eclampsia (PE) and hence is a possible biomarker of PE. It has also been altered in cancers such as glioblastoma, breast carcinoma, and prostate cancer making it an emerging therapeutic target. Among the human HtrAs, HtrA4 is the least characterized protease pertaining to both structure and its functions. Although the members of human HtrA family share a significant structural and functional conservation, subtle structural changes have been associated with certain distinct functional requirements. Therefore, intricate dissection of HtrA4 structural and functional properties becomes imperative to understand its role in various biological and pathophysiological processes. Here, using inter-disciplinary approaches including in silico, biochemical and biophysical studies, we have done a domain-wise dissection of HtrA4 to delineate the roles of the domains in regulating oligomerization, stability, protease activity, and specificity. Our findings distinctly demonstrate the importance of the N-terminal region in oligomerization, stability and hence the formation of a functional enzyme. In silico structural comparison of HtrA4 with other human HtrAs, enzymology studies and cleavage assays with X-linked inhibitor of apoptosis protein (XIAP) show overall structural conservation and allosteric mode of protease activation, which suggest functional redundancy within this protease family. However, significantly lower protease activity as compared with HtrA2 indicates an additional mode of regulation of the protease activity in the cellular milieu. Overall, these studies provide first-hand information on HtrA4 and its interaction with antiapoptotic XIAP thus implicating its involvement in the apoptotic pathway.


Model of age estimation based on dental factors of unknown cadavers among Iranians.
Two hundred and ten cadavers ranging in age from 25 to 60 were studied. Thirty subjects from each five-year interval were selected, and data were collected using the mandibular anterior and premolar teeth after extraction. Sections ranging from 0.5 to 1.0 mm were measured by a stereo microscope with a precision of 0.1 mm. The following factors were determined: attrition, periodontosis, root resorption, secondary dentine apposition, cementum apposition, and translucency of the root. Statistical analysis used the sum of ranks of the dental factors as an independent variable in a linear regression model to estimate the age of the cadaver. Among the different mandibular teeth, the sum of ranks of the first premolar factors had the best correlation coefficient with age. The sum of the dental factors presented a better model than each of the factors alone. The first premolar is recommended as the first step in the estimation of age.


Effects of rumen-protected Capsicum oleoresin on immune responses in dairy cows intravenously challenged with lipopolysaccharide.
The objective of this experiment was to investigate the effects of rumen-protected Capsicum oleoresin (RPC) on productivity and immune responses including feed intake, milk yield and composition, white and red blood cells, lipid peroxidation, and blood concentration of cortisol, haptoglobin, glucose, and insulin in lactating dairy cows experimentally challenged with lipopolysaccharide (LPS). The experiment was a replicated 3 × 3 Latin square design with 9 multiparous Holstein cows in three 28-d periods. Treatments were 0 (control), 100, and 200 mg of RPC/cow per day, mixed with small portions of the total mixed ration and top-dressed. Bacterial LPS was intravenously administered at 1.0 μg/kg of body weight in the last week of each experimental period, and blood samples were collected at 0, 2, 4, 8, and 24 h after administration. Dry matter intake, milk yield, and white blood cells including neutrophils, lymphocytes, monocytes, and eosinophils were decreased, and rectal temperature, hemoglobin, and serum concentrations of cortisol and haptoglobin were increased by LPS. Red blood cells, platelets, and plasma concentration of thiobarbituric acid reactive substances were not affected by LPS. Dry matter intake, milk yield, and milk composition in the 5 d post-LPS challenge were not affected by RPC. Rectal temperature, white blood cells, red blood cells, hemoglobin, and platelets were also not affected by RPC. Compared with the control, RPC tended to decrease cortisol at 2 h following LPS challenge and decreased haptoglobin concentration in serum across sampling points. Concentration of thiobarbituric acid reactive substances in plasma was decreased by RPC at 24 h post-LPS challenge. Glucose and insulin were not affected by RPC, but serum insulin concentration at 8 h was lowered by RPC compared to the control. Collectively, RPC had no or subtle effects on feed intake, milk yield and composition, rectal temperature, white and red blood cells, and serum glucose and insulin concentration in dairy cows challenged by LPS. However, RPC tended to decrease cortisol and decreased concentrations of haptoglobin and thiobarbituric acid reactive substances in blood following LPS challenge. Data suggest that dietary supplementation of RPC may modulate acute phase responses induced by bacterial infection in lactating dairy cows.


Effect of Conditioning and Aging on the Bond Strength and Interfacial Morphology of Glass-ionomer Cement Bonded to Dentin.
To determine the bond stability and the change in interfacial ultrastructure of a conventional glassionomer cement bonded to dentin, with and without pretreatment using a polyalkenoic acid conditioner. The occlusal dentin surfaces of 10 teeth were ground flat. Glass-ionomer cement was bonded to the surfaces either with or without polyalkenoic acid conditioning. The teeth were sectioned into 1-mm2 stick-shaped specimens. The 200 specimens obtained were randomly assigned to four groups with different periods of storage in water: 1 week, 1 month, 3 months, and 6 months. The microtensile bond strength (μTBS) was determined for each storage time. Additional specimens were prepared for transmission electron microscopy (TEM); they were produced with or without prior polyalkenoic acid conditioning in the same manner as for the μTBS test. There was no significant difference in μTBS to conditioned dentin (p > 0.05). After 6 months of aging, the μTBS to non-conditioned dentin was significantly reduced as compared to the 1-week, 1-month, and 3-month results (p < 0.05). The failures appeared to be of a mixed nature, although aging caused more areas of cohesive than adhesive failure in all groups. TEM observation showed a demineralized layer and an amorphous gel phase in the polyalkenoic acid conditioned group. Aging did not reduce the bond strength of the conventional glass-ionomer cement to dentin when the surface was pretreated with a polyalkenoic acid conditioner.


Hemodynamic effects of neurohypophyseal peptides with antidiuretic activity in dogs.
Arginine vasopressin (AVP) is known to produce increases in total peripheral resistance (TPR) and mean arterial pressure (MAP) and decreases in heart rate (HR), cardiac output (CO), and plasma renin activity (PRA). Some recent observations with AVP and synthetic analogues have suggested that under certain conditions, AVP can induce cardiovascular and reninsecretory responses in the opposite directions. To characterize the receptors mediating these responses, the effects of AVP, oxytocin, and synthetic neurohypophyseal analogues with specific antidiuretic, vasoconstrictor, or oxytocic activities were studied in conscious dogs. AVP and 2-phenylalanine-8-ornithine-oxytocin (Phe2Orn8OT, a selective vasoconstrictor agonist) produced similar responses when infused at 10 ng X kg-1 X min-1. That is, TPR and MAP increased, and CO, HR, and PRA decreased. Pretreatment with a selective vasoconstrictor antagonist, [1-(beta-mercapto-beta,beta-cyclopentamethylenepropionic acid) 2-(O-methyl)tyrosine]AVP, abbreviated d(CH2)5Tyr(Me)-AVP (10 micrograms/kg), blocked the actions of Phe2Orn8OT. However, in the presence of d(CH2)5Tyr(Me)AVP, AVP actually decreased TPR and increased CO, HR, and PRA. An analogue with selective antidiuretic activity, 4-valine-8-D-AVP (VDAVP, 10 ng X kg-1 X min-1), produced the same effects as the combination of vasopressin plus d(CH2)5Tyr(Me)AVP. Neither the effects of VDAVP nor of AVP plus antagonist were blocked by propranolol (1 mg/kg). These data indicate that vasopressin, by its antidiuretic activity, produces cardiovascular effects that are opposite to many of those produced by its vasoconstrictor action and that these effects are not dependent on mediation by beta-adrenoceptors.


Transcription factors KLF1 and KLF2 positively regulate embryonic and fetal beta-globin genes through direct promoter binding.
Krüppel-like factors (KLFs) control cell differentiation and embryonic development. KLF1 (erythroid Krüppel-like factor) plays essential roles in embryonic and adult erythropoiesis. KLF2 is a positive regulator of the mouse and human embryonic β-globin genes. KLF1 and KLF2 have highly homologous zinc finger DNA-binding domains. They have overlapping roles in embryonic erythropoiesis, as demonstrated using single and double KO mouse models. Ablation of the KLF1 or KLF2 gene causes embryonic lethality, but double KO embryos are more anemic and die sooner than either single KO. In this work, a dual human β-globin locus transgenic and KLF knockout mouse model was used. The results demonstrate that the human ε- (embryonic) and γ-globin (fetal) genes are positively regulated by KLF1 and KLF2 in embryos. Conditional KO mouse experiments indicate that the effect of KLF2 on embryonic globin gene regulation is at least partly erythroid cell-autonomous. KLF1 and KLF2 bind directly to the promoters of the human ε- and γ-globin genes, the mouse embryonic Ey- and βh1-globin genes, and also to the β-globin locus control region, as demonstrated by ChIP assays with mouse embryonic blood cells. H3K9Ac and H3K4me3 marks indicate open chromatin and active transcription, respectively. These marks are diminished at the Ey-, βh1-, ε- and γ-globin genes and locus control region in KLF1(-/-) embryos, correlating with reduced gene expression. Therefore, KLF1 and KLF2 positively regulate the embryonic and fetal β-globin genes through direct promoter binding. KLF1 is required for normal histone modifications in the β-globin locus in mouse embryos.


Ethical considerations in psychopharmacological research involving children and adolescents.
Increased community utilization of psychotropic medications among children has brought attention to pediatric psychopharmacology research and associated ethical issues. To discuss ethical aspects of child participation in psychopharmacology protocols. Selective review of relevant scientific and regulatory literature. Efficacy and safety of psychotropics in children cannot be entirely inferred from adult data and direct participation of children in research is necessary. Child research must follow special regulations that are in addition to those common to all human research. For research with prospect of direct benefit, a critical factor is whether the risk/benefit ratio is favorable to the participating child. For research without such a prospect, the concepts of minimal risk and minor increase over minimal risk apply. However, the interpretation and application of these principles to specific protocols vary across settings and among ethics committees. Thus far, little empirical investigation has been conducted on children and parents' motivation for research participation, effectiveness of the informed consent and assent procedures, possibility of persistent consequences of exposure to experimental treatments and placebo, and validation of the concepts of minimal risk and minor increase over minimal risk. Research on human subject issues relevant to child participation is a promising approach to improving ethical methods and procedures of pediatric psychopharmacology.


Direct OPTOS Nerve Size Determination of Prevalent Optic Nerve Hypoplasia in Alaska.
Optic nerve hypoplasia (ONH), one of the most common causes of pediatric blindness in developed countries, has been difficult to directly quantify. We sought to measure optic nerve size in Alaskan pediatric patients with optic nerve hypoplasia using ultra-widefield fundus imaging. Adult and pediatric patients underwent conventional ultra widefield fundus imaging (OPTOS, Dunfermline, Scotland) with manual image processing to determine optic nerve size validated against refractive error and nystagmus and compared to optical spectral domain tomography. De-identified cases were then compared relative to visual acuity and birth prevalence. In Alaska's only pediatric ophthalmology outreach clinic, 108 cases of ONH less than 20 years old were clinically identified with 80 having ultra-widefield analysis. Median horizontal optic nerve diameter for 135 normals was 1.70 (95% C.I. 1.49, 2.14) whereas in patients clinically diagnosed with optic nerve hypoplasia was 1.23 (95% C.I 0.38, 1.45). Visual acuity (20/y) was related to horizontal optic nerve diameter (x) by y = 187 x-4.1. Horizontal nerve diameter h could be estimated from vertical nerve diameter v by h = 0.73v + 0.3 even in nystagmus patients. From 108 with ONH, 6 had threshold retinopathy of prematurity, 12 profound nystagmus, 32 legally blind, 6 with septo-optic dysplasia, and 5 with fetal alcohol syndrome. ONH is very prevalent in Alaska occurring at least 8-10 per 10,000 births. Compared to vertical diameter, horizontal diameter was more distinctive of optic nerve hypoplasia and more perturbed by nystagmus. Both were independent of refractive error. When hand-held, spectral domain OCT is not convenient, ultra-widefield fundus analysis is recommended for direct estimation of optic nerve size in children and adults. Optic nerve hypoplasia is prevalent in Alaskan children.


Association Between Breastfeeding and Child Stunting in Mexico.
Globally, the prevalence of child stunting has been decreasing over the past decades. However, in low- and middle-income countries such as Mexico, stunting is still the most prevalent form of undernutrition affecting a large number of children in the most vulnerable conditions. Breastfeeding has been identified as one of the key affordable and modifiable maternal health behaviors protecting against child stunting. To examine the association between breastfeeding (defined as never breastfed, any breastfeeding for <6 months, and any breastfeeding for ≥6 months) and other individual-, household-, and area-level factors with child stunting (defined as length/height-for-age-z-score for sex under -2 standard deviations of the World Health Organization child growth standards' median) in Mexico. Secondary data analysis using the 2012 Mexican Health and Nutrition Survey, which allowed representativeness of rural and urban areas at national level and among 4 regions in Mexico. Our subset included data on 2,089 singleton Mexican children aged 6-35 months with information on previously identified risk and protective factors for stunting. We conducted fixed- and mixed-effects logistic regression models sequentially controlling for each level of factors. Overall, 12.3% of children were stunted and 71.1% were breastfed for ≥6 months. Any breastfeeding and being female were consistent protective factors against child stunting across all models. In contrast, child low birthweight, maternal short stature, higher number of children aged <5 years per household, and moderate to severe food insecurity were consistent risk factors for child stunting across all models. According to our findings, efforts to reduce child stunting in Mexico should include prenatal strategies aiming to prevent low birthweight offspring particularly among short-stature women, moderate to severe food insecure households, families with a higher number of children aged <5 years, and indigenous communities. Postnatal components should include multilevel strategies to support breastfeeding.