Refining the study of decision-making in animals: differential effects of d-amphetamine and haloperidol in a novel touchscreen-automated Rearing-Effort Discounting (RED) task and the Fixed-Ratio Effort Discounting (FRED) task.
Effort-based decision-making is impaired in multiple psychopathologies leading to significant impacts on the daily life of patients. Preclinical studies of this important transdiagnostic symptom in rodents are hampered, however, by limitations present in currently available decision-making tests, including the presence of delayed reinforcement and off-target cognitive demands. Such possible confounding factors can complicate the interpretation of results in terms of decision-making per se. In this study we addressed this problem using a novel touchscreen Rearing-Effort Discounting (RED) task in which mice choose between two single-touch responses: rearing up to touch an increasingly higher positioned stimulus to obtain a High Reward (HR) or touching a lower stimulus to obtain a Low Reward (LR). To explore the putative advantages of this new approach, RED was compared with a touchscreen version of the well-studied Fixed Ratio-based Effort Discounting (FRED) task, in which multiple touches are required to obtain an HR, and a single response is required to obtain an LR. Results from dopaminergic (haloperidol and d-amphetamine), behavioral (changes in the order of effort demand; fixed-ratio schedule in FRED or response height in RED), and dietary manipulations (reward devaluation by pre-feeding) were consistent with the presence of variables that may complicate interpretation of conventional decision-making tasks, and demonstrate how RED appears to minimize such variables.


Locating helicopter emergency medical service bases to optimise population coverage versus average response time.
New South Wales (NSW), Australia has a network of multirole retrieval physician staffed helicopter emergency medical services (HEMS) with seven bases servicing a jurisdiction with population concentrated along the eastern seaboard. The aim of this study was to estimate optimal HEMS base locations within NSW using advanced mathematical modelling techniques. We used high resolution census population data for NSW from 2011 which divides the state into areas containing 200-800 people. Optimal HEMS base locations were estimated using the maximal covering location problem facility location optimization model and the average response time model, exploring the number of bases needed to cover various fractions of the population for a 45 min response time threshold or minimizing the overall average response time to all persons, both in green field scenarios and conditioning on the current base structure. We also developed a hybrid mathematical model where average response time was optimised based on minimum population coverage thresholds. Seven bases could cover 98% of the population within 45mins when optimised for coverage or reach the entire population of the state within an average of 21mins if optimised for response time. Given the existing bases, adding two bases could either increase the 45 min coverage from 91% to 97% or decrease the average response time from 21mins to 19mins. Adding a single specialist prehospital rapid response HEMS to the area of greatest population concentration decreased the average state wide response time by 4mins. The optimum seven base hybrid model that was able to cover 97.75% of the population within 45mins, and all of the population in an average response time of 18 mins included the rapid response HEMS model. HEMS base locations can be optimised based on either percentage of the population covered, or average response time to the entire population. We have also demonstrated a hybrid technique that optimizes response time for a given number of bases and minimum defined threshold of population coverage. Addition of specialized rapid response HEMS services to a system of multirole retrieval HEMS may reduce overall average response times by improving access in large urban areas.


Predicting efficacy and guiding procedure choice in non-vascularized bone grafting: a CT Radiomics and clinical predictor approach.
There is no practical approach for accurately predicting the efficacy of non-vascularized bone grafting (NVBG) and guiding its optimal procedure. This study enrolled 153 patients with 182 hips that underwent NVBG procedures. The patients were randomly divided into a training cohort (n = 130) and a validation cohort (n = 52). In the training cohort, radiomics model, clinical model, and combined radiomics-clinical (C-R) model were constructed using Rad-scores and clinical predictors to predict the efficacy of NVBG. The optimal model was visualized by a nomogram and assessed by decision curve analysis (DCA). 128 hips that underwent successful NVBG were then randomized into a new training cohort (n = 92) and a new validation cohort (n = 36), and three models were constructed and validated to predict the choice of NVBG procedure. Japanese Investigation Committee (JIC) classification, exposure to risk factors postoperative, and Rad-scores consisting of four radiomics features were independent predictors for the efficacy of NVBG (P < 0.05). The C-R model provided better performance in both the training cohort (AUC: 0.818) and validation cohort (AUC: 0.747). To predict the choice of NVBG procedure, the C-R model built by JIC classification and Rad-scores consisting of five radiomics features showed the finest performance in both cohorts (AUC: 0.860 and 0.800, respectively). DCA showed great benefit using the C-R model for the choice of NVBG procedure. The approach integrated by CT radiomics and clinical predictors can be visually and quantitatively applied to predict the efficacy and guide the choice of NVBG procedure with great predictive accuracy.


Assessment of resistance of nasal continuous positive airway pressure interfaces.
To compare the resistance of interfaces used for the delivery of nasal continuous positive airway pressure (CPAP) in neonates, as measured by the generated system pressure at fixed gas flows, in an in vitro setting. Gas flows of 6, 8 and 10 L/min were passed through three sizes of each of a selection of available neonatal nasal CPAP interfaces (Hudson prong, RAM Cannula, Fisher & Paykel prong, Infant Flow prong, Fisher & Paykel mask, Infant Flow mask). The expiratory limb was occluded and pressure differential measured using a calibrated pressure transducer. Variation in resistance, assessed by mean pressure differential, was seen between CPAP interfaces. Binasal prong interfaces typically had greater resistance at the smallest assessed sizes, and with higher gas flows. However, Infant Flow prongs produced low pressures (<1.5 cmH2O) at all sizes and gas flows. RAM Cannula had a high resistance, producing a pressure >4.5 cmH2O at all sizes and gas flows. Both nasal mask interfaces had low resistance at all assessed sizes and gas flows, with recorded pressure <1 cmH2O in all cases. There is considerable variation in measured resistance of available CPAP interfaces at gas flows commonly applied in clinical neonatal care. Use of interfaces with high resistance may result in a greater drop in delivered airway pressure in comparison to set circuit pressure, which may have implications for clinical efficacy. Device manufacturers and clinicians should consider CPAP interface resistance prior to introduction into routine clinical care.


Likelihood ratio of sonographic findings in discriminating hydrosalpinx from other adnexal masses.
The objective of our study was to describe the "waist" sign as a feature of hydrosalpinx and to calculate the likelihood ratio of sonographic findings for predicting that a cystic adnexal mass is a hydrosalpinx. Consecutive institutional records were reviewed to identify patients with surgically excised cystic adnexal masses and preoperative sonograms that had been used to identify the mass. Two radiologists blinded to outcome reviewed the sonograms and recorded the presence of five specific sonographic findings: incomplete septation, short linear projection, small round projection, tubular shape, and presence of a waist. The likelihood ratios of these signs and combinations of signs for the diagnosis of hydrosalpinx were calculated. There were 55 patients with 67 cystic adnexal masses. Of the 67 cystic adnexal masses, there were 26 hydrosalpinges (39%), 36 cystic ovarian neoplasms (54%), and five paraovarian masses (7%). The likelihood ratios for the sonographic findings were as follows: incomplete septation, 2.1; short linear projection, 3.5; small round projection, 2.7; tubular shape, 10.5; and waist sign, 20.5. The waist sign in combination with tubular shape was found in 12 hydrosalpinges and no other masses (likelihood ratio of between 18.9 and infinity). Small round projection combined with tubular shape was found in 14 hydrosalpinges and one other mass (likelihood ratio of 22.1). Hydrosalpinx can be diagnosed with the highest likelihood when a tubular mass with the waist sign or a tubular mass with small round projections is encountered. Incomplete septations and short linear projections are less discriminating findings of hydrosalpinx.


Altered response to partial bladder outlet obstruction in mice lacking inducible nitric oxide synthase.
Following prolonged partial bladder outlet obstruction (BOO) in the mouse, cholinergic mediated detrusor contractility decreases. Previous work has demonstrated an increase in the inducible form of nitric oxide synthase (iNOS) at the mRNA and protein levels soon after obstruction. Since nitric oxide (NO), the product of the action of iNOS on molecular oxygen and l-arginine, produces vasodilation and decreases platelet aggregation, we believe it is an integral part of the initial detrusor response to obstruction. These experiments evaluated the detrusor response in mice incapable of producing iNOS. Wild type and knockout mice were partially obstructed for 1, 3, and 5 weeks. Physiologic evaluation consisted of cystometric analyses, and muscle strip studies in response to cholinergic and electrical stimulation. Strips were also relaxed with L-arginine, sodium nitroprusside, and 8-bromoguanosine 3' - 5' cyclic GMP, after precontraction. After 5 weeks of obstruction, both wild type (WT) and knockout (KO) mouse bladders increased significantly in weight. WT bladders obstructed for 5 weeks had the greatest capacity (increase of 42%, p = 0.022), and a decreased contractile response to carbachol (decrease of 32% at 10-5 M, p = 0.018). No differences were noted at 1 and 3 weeks of obstruction. In contrast, KO mice had a significantly larger bladder capacity at 1 week of obstruction compared with WT, and had significantly lower responses to electrical stimulation than WT at the same time (p = 0.03). Additionally, after 5 weeks of obstruction, bladder capacity and contractility returned to baseline levels in KO mice, at a time when WT mice had significantly larger capacity and decreased contractility. Bladder function following partial BOO in mice incapable of producing iNOS differed significantly from the normal response. Our data suggest that generation of iNOS soon after obstruction is necessary to prevent detrusor dysfunction at that time. Moreover, the enhanced function seen in KO bladders after longer periods of obstruction (5 weeks) in comparison to WT bladders suggests that reactive nitrogen species-induced protein nitrosylation may be involved in the loss of contractile function observed after more prolonged periods of obstruction.


Development of a surgical navigation system based on 3D Slicer for intraoperative implant placement surgery.
Implant placement has been widely used in various kinds of surgery. However, accurate intraoperative drilling performance is essential to avoid injury to adjacent structures. Although some commercially-available surgical navigation systems have been approved for clinical applications, these systems are expensive and the source code is not available to researchers. 3D Slicer is a free, open source software platform for the research community of computer-aided surgery. In this study, a loadable module based on Slicer has been developed and validated to support surgical navigation. This research module allows reliable calibration of the surgical drill, point-based registration and surface matching registration, so that the position and orientation of the surgical drill can be tracked and displayed on the computer screen in real time, aiming at reducing risks. In accuracy verification experiments, the mean target registration error (TRE) for point-based and surface-based registration were 0.31±0.06mm and 1.01±0.06mm respectively, which should meet clinical requirements. Both phantom and cadaver experiments demonstrated the feasibility of our surgical navigation software module.


The pathogenesis of choroidal neovascularization in patients with age-related macular degeneration.
Laser photocoagulation and several experimental treatments for choroidal neovascularization (CNV) in patients with age-related macular degeneration attempt to ablate the neovascularization, but do not address underlying angiogenic stimuli. As a result, recurrences are a major problem. Drug treatment to counter the growth of CNV would be a major advance, but its development is impeded by lack of knowledge concerning the stimuli and other molecular signals involved in the pathogenesis of CNV. Herein we explore clues that can be gleaned from clinical, epidemiological, pathological, and experimental data. These suggest that abnormalities of the extracellular matrix of retinal pigmented epithelial (RPE) cells may promote a pro-angiogenic RPE phenotype that contributes to the development of CNV. This provides a general hypothesis that can be tested, but it is also necessary to test hypotheses regarding the specific alterations in gene expression that contribute to CNV. Identification of alterations in gene expression will provide targets for rational design of drug treatment.


Determination of hydrastine and berberine in goldenseal raw materials, extracts, and dietary supplements by high-performance liquid chromatography with UV: collaborative study.
A multilaboratory collaborative study was conducted on a high-performance liquid chromatographic (HPLC) method utilizing UV detection, previously validated using AOAC single-laboratory validation guidelines for determination of hydrastine and berberine in goldenseal (Hydrastis canadensis L.) raw materials, extracts, and dietary supplements at levels ranging from 0.4 to 6% (w/w). Nine collaborating laboratories determined the hydrastine and berberine content in 8 blind samples. Sample materials included powdered botanical raw materials, whole root material, and 4 finished product dietary supplements containing either goldenseal powdered root material or extract. The materials were extracted with an acidified water and acetonitrile solution. HPLC analyses of the extracts were performed on a C18 column using UV detection at 230 nm. Results for powdered root material and capsule products ranged from about 0.2% (w/w) for each alkaloid to about 4% (w/w) for each alkaloid. Liquid tincture results were approximately 4000-5000 microg/mL for each alkaloid. Reproducibility relative standard deviations (RSDR) for hydrastine ranged from 2.68 to 6.65%, with HorRat values ranging from 0.77 to 1.89. RSDR for berberine ranged from 5.66 to 7.68%, with HorRat values ranging from 1.32 to 2.12. All finished products containing goldenseal extract yielded HorRat values <2.0. Based on these results, the method is recommended for Official First Action for determination of hydrastine and berberine in goldenseal raw materials and dietary supplement finished products containing powdered goldenseal and goldenseal extract.


Profile of the patients who present to immunology outpatient clinics because of frequent infections.
We aimed to determine the rate of primary immune deficiency (PID) among children presenting to our immunology outpatient clinic with a history of frequent infections and with warning signs of primary immune deficiency. The files of 232 children aged between 1 and 18 years with warning signs of primary immune deficiency who were referred to our pediatric immunology outpatient clinic with a complaint of frequent infections were selected and evaluated retrospectively. Thirty-six percent of the subjects were female (n=84) and 64% were male (n=148). PID was found in 72.4% (n=164). The most common diagnosis was selective IgA deficiency (26.3%, n=61). The most common diseases other than primary immune deficiency included reactive airway disease and/or atopy (34.4%, n=22), adenoid vegetation (12.3%, n=8), chronic disease (6.3%, n=4) and periodic fever, aphtous stomatitis and adenopathy (4.6%, n=3). The majortiy of the subjects (90.5%, n=210) presented with a complaint of recurrent upper respiratory tract infection. PID was found in all subjects who had bronchiectasis. The rates of the diagnoses of variable immune deficiency and Bruton agammaglubulinemia (XLA) were found to be significantly higher in the subjects who had lower respiratory tract infection, who were hospitalized because of infection and who had a history of severe infection compared to the subjects who did not have these properties (p<0.05 and p<0.01, respectively). Growth and developmental failure was found with a significantly higher rate in the patients who had a diagnosis of severe combined immune deficiency or hyper IgM compared to the other subjects (p<0.01). No difference was found in the rates of PID between the age groups, but the diagnosis of XLA increased as the age of presentation increased and this was considered an indicator which showed that patients with XLA were being diagnosed in a late period. It was found that the rate of diagnosis was considerably high (72.4%), when the subjects who had frequent infections were selected by the warning signs of PID.


Drug-metabolising enzyme polymorphisms and predisposition to anti-tuberculosis drug-induced liver injury: a meta-analysis.
Although some case-control studies have investigated the association between drug-metabolising enzyme (DME) gene polymorphisms and susceptibility to anti-tuberculosis drug-induced liver injury (ATLI), their results are conflicting, mainly due to limited power. To review the literature systematically, by means of a meta-analytical review, to evaluate the putative association and provide a quantitative summary estimate on the association with ATLI. We searched the databases of MEDLINE, PubMed, EMBASE and CBMdisc from 1966 to May 2007 using 'DME', 'hepatotoxicity', 'genetic polymorphism', 'genetic susceptibility' in combination with 'antitubercular agents', performed a manual search of citations from relevant original studies and review articles, and corresponded with authors. Nine eligible articles were included in this meta-analysis, including five on N-acetyltransferase 2 (NAT2), four on cytochrome P450 2E1 (CYP2E1) and two on glutathione S-transferase (GST) studies, separately. The overall ORs of ATLI risk associated with NAT2 homozygous variant genotype (mt/mt), CYP2E1 homozygous wild genotype (*1A/*1A), GSTM1 homozygous null genotype (null/null) and GSTT1 homozygous null genotype (null/null) were respectively 1.93 (95%CI 0.81-4.62), 2.22 (95%CI 1.06-4.66), 2.62 (95%CI 1.45-4.75) and 1.18 (95%CI 0.61-2.29). In addition, the OR for Asian ATLI associated with the NAT2 homozygous variant (mt/mt) and the combined genotype (w/w + w/mt) was 2.52 (95%CI 1.49-4.26). NAT2 mt/mt, CYP2E1*1A/*1A and GSTM1 null/null were observed to increase the risk of ATLI in tuberculosis patients. Our results support the hypothesis that NAT2 mt, CYP2E1*1A and GSTM1 null have a modest effect on genetic susceptibility to ATLI, but no significant evidence for GSTT1 null/null.


Long-Lasting, Pathway-Specific Impairment of a Novel Form of Spike-Timing-Dependent Long-Term Depression by Neuropathic Pain in the Anterior Cingulate Cortex.
Malfunctioning synaptic plasticity is one of the major mechanisms contributing to the development of chronic pain. We studied spike-timing dependent depression (tLTD) in the anterior cingulate cortex (ACC) of male mice, a brain region involved in processing emotional aspects of pain. tLTD onto layer 5 pyramidal neurons depended on postsynaptic calcium-influx through GluN2B-containing NMDARs and retrograde signaling via nitric oxide to reduce presynaptic release probability. After chronic constriction injury of the sciatic nerve, a model for neuropathic pain, tLTD was rapidly impaired; and this phenotype persisted even beyond the time of recovery from mechanical sensitization. Exclusion of GluN2B-containing NMDARs from the postsynaptic site specifically at projections from the anterior thalamus to the ACC caused the tLTD phenotype, whereas signaling downstream of nitric oxide synthesis remained intact. Thus, transient neuropathic pain can leave a permanent trace manifested in the disturbance of synaptic plasticity in a specific afferent pathway to the cortex.SIGNIFICANCE STATEMENT Synaptic plasticity is one of the main mechanisms that contributes to the development of chronic pain. Most studies have focused on potentiation of excitatory synaptic transmission, but very little is known about the reduction in synaptic strength. We have focused on the ACC, a brain region associated with the processing of emotional and affective components of pain. We studied spike-timing dependent LTD, which is a biologically plausible form of synaptic plasticity, that depends on the relative timing of presynaptic and postsynaptic activity. We found a long-lasting and pathway-specific suppression of the induction mechanism for spike-timing dependent LTD from the anterior thalamus to the ACC, suggesting that this pathology might be involved in altered emotional processing in pain.


Blue light cystoscopy with hexylaminolevulinate: Our 7 years experience.
The objective of the present study is to evaluate the diagnostic accuracy of hexylaminolevulinate (HAL) blue light cystoscopy compared with standard white light cystoscopy (WLC) in daily practice. An observational, comparative, controlled (within patient) study was carried out at our Center. 61 consecutive patients with suspected or confirmed bladder cancer were recruited for the study from January 2008 until January 2015. Patients with suspected bladder cancer (positive cytology with negative WLC) or history of previous high-grade NMIBC or CIS were included in the study. Biopsies/resection of each positive lesion/suspicious areas were always taken after the bladder was inspected under WLC and BLC. Diagnoses of bladder tumor or CIS were considered as positive results, and the presence of normal urothelium in the biopsy specimen as negative result. 61 BLC were performed. 15/61 (24.5%) with suspected initial diagnosis of NMIBC and 46/61 (75.5%) with a history of high-risk non-muscle invasive bladder cancer (NMIBC). We performed a total of 173 biopsies/TURBT of suspicious areas: 129 positive only to the BLC and 44 both positive to WLC and BLC. 84/173 biopsies/TURBT were positive for cancer. All 84 NMIBC were positive to the BLC, while 35/84 were positive to the WLC with a sensitivity of BLC and WLC respectively of 100% and 41.7%. Sensitivity of WLC for highgrade NMIBC and CIS was 34.1% and 39% respectively while sensitivity of BLC for high-grade NMIBC and CIS was 100%. The specificity of the WLC was 79.9% compared to 48.5% of the BLC. The positive predictive value of BLC and WLC were respectively 48% (95% CI: 0.447-0.523) and 79% (95% CI: 0.856-0.734). Our data confirm those reported in the literature: BLC increases the detection rate of NMIBC particularly in high risk patients (history of CIS or high grade). BLC is a powerful diagnostic tool in the diagnosis of bladder cancer if malignancy is suspected (positive urine cytology) and if conventional WLC is negative.


Structure-function analysis reveals the molecular determinants of the impaired biological function of DAX-1 mutants in AHC patients.
Mutations in the DAX-1 (NR0B1) gene cause the X-linked form of adrenal hypoplasia congenita (AHC), which is constantly found associated with hypogonadotropic hypogonadism (HHG). DAX-1 encodes an atypical orphan member of the nuclear hormone receptor superfamily. DAX-1 acts at multiple levels to repress the expression of genes involved in steroid hormone metabolism through a potent transcriptional repression domain present in its C-terminus, which is similar to the nuclear receptors' ligand binding domain. All DAX-1 mutations causing AHC/HHG alter the protein C-terminal domain, impairing its nuclear localization and, consequently, its transcriptional repression activity. Here we show that DAX-1 AHC mutants have a misfolded conformation, which correlates with their cytoplasmic retention. Extensive structure-function analysis reveals that the chemical nature of amino acid residues at positions interested by AHC mutations and critical determinants in helix 12 affect DAX-1 nuclear localization and transcriptional silencing. Surprisingly, mutations in a conserved putative corepressor binding surface have a negative effect upon DAX-1 transcriptional repression only when they also affect protein expression levels. These data suggest that a folding defect underlies the impaired function of DAX-1 missense mutants found in AHC/HHG patients and that interactions with transcriptional cofactors different from known corepressors mediate DAX-1 silencing properties.


Possible mechanistic insights into iron homeostasis role of the action of 4-aminoquinolines (chloroquine/hydroxychloroquine) on COVID-19 (SARS-CoV-2) infection.
With the recent direction in drug repurposing, many approved drugs have been evaluated to assess their effect on the coronavirus or SARS-CoV-2 infection (COVID-19). Driving this path, chloroquine (CQ) has been used in the treatment of malaria and hydroxychloroquine (HCQ) in immunomodulatory and anti-thrombotic action, playing a leading role in initial management of the viral infection. Literature search was done using Google Scholar, PubMed and Scopus database using keywords "chloroquine" "SARS-CoV-2" "COVID-19" "mechanism of action" and articles of interest were selected providing evidence of the possible role of CQ in viral infection. In a bid to understand how and if CQ and HCQ would exert their anti-viral property, mechanistic exegesis was done to review various proposed mechanisms of action. This revealed the inhibition of viral attachment and entry, inhibition of enveloped glycoprotein, inhibition of the development and proliferation of new viral particles as the way they perform their action. There is an interplay between iron metabolism and homeostasis with COVID-19 infection and viral reproduction. This study aims to show the functional role of CQ and HCQ, as well as to provide possible mechanistic insight on the role of iron on viral infection, iron starvation and its downstream cellular pathways involving hepcidin and proinflammatory cytokines. The overall aim of providing possible mode of action of CQ and HCQ in the management of COVID-19 infection is exhibited via its anti-viral, anti-inflammatory and anti-thrombotic activities.


Exploring the picosecond time domain of the solvation dynamics of coumarin 153 within beta-cyclodextrins.
We report molecular dynamics simulation results of equilibrium and dynamical characteristics pertaining to the solvation of the dye coumarin 153 (C153) trapped within hydrophobic cavities of di- and trimethylated beta-cyclodextrins (CD) in aqueous solutions. We found that stable configurations of the encapsulated probe are characterized by a slanted docking, in which the plane of the C153 lies mostly parallel to one of the glucose units of the CD. "In and out" dynamical modes of the encapsulated probe present very small amplitudes. The rotational dynamics of the trapped coumarin can be cast in terms of a simple model that includes diffusive motions within a local restrictive environment coupled to the overall rotational motion of the CD. We have examined the early stages of the solvation response of the environment following a vertical excitation of the probe. Regardless of the degree of CD methylation, the water dynamical response seems to be completed within 2-3 ps and does not differ substantially from that observed for nonencapsulated probes. The CD response is characterized by a single, subpicosecond relaxation that involves intramolecular motions. We also explored dynamical modes that could account for the recently reported persistence of Stokes shifts in the nanosecond time domain. In all cases, the only sources of ultraslow dynamics that we detected were those associated with gauche-trans interconversions in primary hydroxyl chains of the CD, which do not seem to be directly connected to the electronic excitation of the probe.


Studies of the structure of lung surfactant protein SP-A.
SP-A, a glycoprotein of pulmonary surfactant, consists of an NH2-terminal domain containing a collagen-like sequence and a COOH-terminal domain with sequence homology to several Ca2(+)-dependent lectins. We have compared the size, thermal stability, and secondary structure of recombinant SP-A, the product of a fibroblast line transfected with a single human gene encoding SP-A, with natural SP-A isolated from canine and human lungs. Our results suggest both recombinant and natural SP-A are assembled as large oligomers. More variability in the degree of oligomerization was observed with recombinant human SP-A than with natural canine SP-A. As shown by collagenase digestion, the full assembly of protein subunits was dependent on an intact collagen-like domain. The cysteines in the noncollagen domain of SP-A form intrachain bonds between residues 135-226 and 204-218. The circular dichroism spectra of both recombinant and natural SP-A were consistent with the presence of a collagen-like triple helix. As determined by the change in ellipticity at 205 nm, the thermal transition temperatures of canine, natural human, and recombinant SP-A were 51.5, 52.3, and 42.0 degrees C, respectively. These results suggest differences in the assembly and stability of the natural and recombinant proteins.


The importance of the CXCL12-CXCR4 chemokine ligand-receptor interaction in prostate cancer metastasis.
Chemokines or chemotactic cytokines are known to be important in the directional migration or chemotaxis of leucocytes in conditions of homeostasis and in inflammatory or immunological responses. However, the role of chemokines is extending beyond their involvement in mediating leucocyte trafficking with an increasing body of evidence suggesting these proteins are intimately involved in many stages of tumour development and progression. Our aim was to study the role of the CXCL12:CXCR4 chemokine ligand:receptor complex in determining the organ-specific metastasis of prostate cancer. CXCR4 mRNA expression was determined by RT-PCR in 3 metastatic prostate cancer cell lines DU145, LNCaP and PC3, the primary prostate cancer cell line 1542 CPT3X and the normal prostate epithelial cell lines 1542 NPTX and Pre 2.8. This was followed by Taqman quantitative PCR analysis of CXCR4 mRNA in these cell lines. Flow cytometry analysis was then used to measure the expression of the CXCR4 receptor protein on the cell surface. The influence of the receptor on cell migration was studied using Transwell, Migration Assays. Finally, Taqman quantitative PCR was performed on RNA obtained from laser microdissected fresh primary prostate tumour and benign tissue samples from patients. In DU145, LNCaP and PC3 CXCR4 mRNA expression was approximately 1000, 400 and 21 times respectively that of 1542 NPTX, Pre 2.8 and 1542 CPT3X. In patient primary tumour samples and patient benign tissue specimens CXCR4 mRNA expression was similar to that of the metastatic cell line DU145. Flow cytometry analysis showed that significantly higher levels of the CXCR4 receptor were present on the cell surface of the 3 metastatic cell lines. Migration studies revealed that chemotaxis of the metastatic cell lines PC3 and DU145 was enhanced by CXCL12 ligand and inhibited by antibody to CXCR4. CXCL12 did not influence the migration of the normal prostate epithelial cell line 1542 NPTX. We have demonstrated that human prostate cell lines derived from metastases express functional CXCR4 receptor and that CXCL12 ligand enhances their migratory capabilities. Also, laser microdissected primary patient tumours and patient benign tissue specimens express CXCR4 mRNA at high levels (it is suggested that post-transcriptional modification of the CXCR4 receptor plays a major role in regulating protein expression). These results suggest prostate cancers may be influenced by the CXCL12:CXCR4 pathway during metastasis. This pathway would provide a novel target for therapeutic intervention.


Comparing the levels of CTLA-4-dependent biological defects in patients with LRBA deficiency and CTLA-4 insufficiency.
Lipopolysaccharide-responsive beige-like anchor protein (LRBA) deficiency and cytotoxic T-lymphocyte protein-4 (CTLA-4) insufficiency are recently described disorders that present with susceptibility to infections, autoimmunity, and lymphoproliferation. Clinical and immunological comparisons of the diseases with long-term follow-up have not been previously reported. We sought to compare the clinical and laboratory manifestations of both diseases and investigate the role of flow cytometry in predicting the genetic defect in patients with LRBA deficiency and CTLA-4 insufficiency. Patients were evaluated clinically with laboratory assessments for lymphocyte subsets, T follicular helper cells (TFH ), LRBA expression, and expression of CD25, FOXP3, and CTLA4 in regulatory T cells (Tregs) at baseline and 16 h post-stimulation. LRBA-deficient patients (n = 29) showed significantly early age of symptom onset, higher rates of pneumonia, autoimmunity, chronic diarrhea, and failure to thrive compared to CTLA-4 insufficiency (n = 12). In total, 29 patients received abatacept with favorable responses and the overall survival probability was not different between transplanted versus non-transplanted patients in LRBA deficiency. Meanwhile, higher probability of survival was observed in CTLA-4-insufficient patients (p = 0.04). The T-cell subsets showed more deviation to memory cells in CTLA-4-insufficiency, accompanied by low percentages of Treg and dysregulated cTFH cells response in both diseases. Cumulative numbers of autoimmunities positively correlated with cTFH frequencies. Baseline CTLA-4 expression was significantly diminished in LRBA deficiency and CTLA-4 insufficiency, but significant induction in CTLA-4 was observed after short-term T-cell stimulation in LRBA deficiency and controls, while this elevation was less in CTLA-4 insufficiency, allowing to differentiate this disease from LRBA deficiency with high sensitivity (87.5%) and specificity (90%). This cohort provided detailed clinical and laboratory comparisons for LRBA deficiency and CTLA-4 insufficiency. The flow cytometric approach is useful in predicting the defective gene; thus, targeted sequencing can be conducted to provide rapid diagnosis and treatment for these diseases impacting the CTLA-4 pathway.


Immunosuppressive mechanisms in protein-calorie malnutrition.
Protein-calorie malnutrition (PCM) induces immunosuppression leading to increased mortality rates. Impaired macrophage respiratory burst activity (superoxide anion [O2-] generation) occurs in PCM, but cellular mechanisms are unclear. The major pathway resulting in O2- production involves inositol lipid-dependent signal transduction. This study examined the effect of mild versus severe PCM on macrophage O2- generating signal transduction pathways specific for responses to Candida albicans. Mice (CFW/Swiss Webster: n = 300) were randomized to either control or low protein diets for 3 or 8 weeks. Peritoneal macrophages were harvested for O2- production, mannose-fucose receptor (MFR) expression, membrane phospholipid analysis, arachidonic acid (AA) content, prostaglandin E2 (PGE2) production, and protein kinase C levels. O2- release was impaired in both mild and severe PCM. MFR expression was also decreased at these time points. Inositol lipid content was significantly lower at the 8-week time point only, although PGE2 and AA were significantly higher in the low protein diet group at 3 weeks. Protein kinase C levels were unchanged by PCM. Thus, mild PCM significantly increases macrophage-PGE2 production secondary to increased AA phospholipid content, with subsequent inhibition of O2- and MFR expression. Severe PCM inhibits macrophage (O2-) through depletion of critical membrane phospholipid components with subsequent impairment in signal transduction.


Antidepressants for depression in medical illness.
To determine whether antidepressants are clinically effective and acceptable for the treatment of depression in people who also have a physical illness. Medline, Cochrane Library Trials Register and Cochrane Depression and Neurosis Group Trials Register were all systematically searched, supplemented by hand searches of two journals and reference searching. All relevant randomised trials comparing any antidepressant drug (as defined in the British National Formulary) with placebo or no treatment, in patients of either sex over 16, who have been diagnosed as depressed by any criterion, and have a specified physical disorder (for example cancer, myocardial infarction). "Functional" disorders where there is no generally agreed physical pathology (e.g. irritable bowel syndrome) were excluded. The main outcome measures are numbers of individuals who recover/improve at the end of the trial and, as a proxy for treatment acceptability, numbers who complete treatment. Data was extracted independently by the reviewers onto data collection forms and differences settled by discussion. 18 studies were included, covering 838 patients with a range of physical diseases (cancer 2, diabetes 1, head injury 1, heart 1, HIV 5, lung 1, multiple sclerosis 1, renal 1, stroke 3, mixed 2). Depression was diagnosed clinically in 3 studies, otherwise by structured interview or checklist. Only 5 studies described how they performed randomisation. 1 study compared drug with no treatment, and the rest with placebo: all of the latter said they were double blind. 6 studies used SSRIs, 3 atypical antidepressants, and the remainder tricyclics. Patients treated with antidepressants were significantly more likely to improve than those given placebo (13 studies, OR 0.37, 95% CI 0.27-0.51) or no treatment (1 study, OR 3.45, 95% CI 11.1-1.10). About 4 patients would need to be treated with antidepressants to produce one recovery from depression which would not have occurred had they been given placebo (NNT 4.2, 95% CI 3.2-6.4). Most antidepressants (tricyclics and SSRIs together, 15 trials ) produced a small but significant increase in dropout (OR 1.66, 95% CI 1.14-2.40. NNH 9.8, 95% CI 5.4-42.9). The "atypical" antidepressant mianserinproduced significantly less dropout than placebo. Only 2 studies used numerical scales designed to measure effects on function and quality of life; in HIV (Karnofsky scale), drug was better than no treatment; in lung disease (Sickness Impact Profile), drug was not significantly different from placebo. Only 7 studies reported looking for changes in the physical disease. Antidepressants produced no change in immune function in HIV relative to placebo (2 studies) or no treatment (1 study). Relative to placebo, antidepressants produced no change in cardiovascular function in heart disease, in respiratory function in lung disease, or in vital signs or laboratory tests in cancer (1 study each). Nortriptyline produced worse control in diabetes. Trends towards tricyclics being more effective than SSRIs, but also more likely to produce dropout were noted, but these are based on non-randomised comparisons between trials. The review provides evidence that antidepressants, significantly more frequently than either placebo or no treament, cause improvement in depression in patients with a wide range of physical diseases. About 4 patients would need to be treated with antidepressants to produce one recovery from depression which would not have occurred had they been given placebo (NNT 4.2, 95% CI 3.2-6.4). Antidepressants seem reasonably acceptable to patients, in that about 10 patients would need to be treated with antidepressants to produce one dropout from treatment which would not have occurred had they been given placebo (NNH 9.8, 95% CI 5.4-42.9). The evidence is consistent across the trials, apart from 2 trials in cancer, where the "atypical" antidepressant mianserin produced significantly less dropout than placebo. Trends towards tricyclics being more effective than SSRIs, but also more likely to produce dropout were noted, but these are based on non-randomised comparisons between trials. Problems with the evidence include most of the trials' use of observers, rather than patients, to decide on improvement, and concentration mainly on symptoms rather than function and quality of life. There is also a possibility of undetected negative trials. Nevertheless, the review provides evidence that use of antidepressants should at least be considered in those with both physical illness and depression. Regarding diagnosis, the existence of a cheap and readily available treatment for depression should encourage detailed assessment of persistent low mood in the physically ill.


Complementation of deletion mutants in the genes encoding the F1-ATPase by expression of the corresponding bovine subunits in yeast S. cerevisiae.
The F1F0 ATP synthase is composed of the F1-ATPase which is bound to F0, in the inner membrane of the mitochondrion. Assembly and function of the enzyme is a complicated task requiring the interactions of many proteins for the folding, import, assembly, and function of the enzyme. The F1-ATPase is a multimeric enzyme composed of five subunits in the stoichiometry of alpha3beta3gammadeltaepsilon. This study demonstrates that four of the five bovine subunits of the F1-ATPase can be imported and function in an otherwise yeast enzyme effectively complementing mutations in the genes encoding the corresponding yeast ATPase subunits. In order to demonstrate this, the coding regions of each of the five genes were separately deleted in yeast providing five null mutant strains. All of the strains displayed negative or a slow growth phenotype on medium containing glycerol as the carbon source and strains with a null mutation in the gene encoding the gamma-, delta- or epsilon-gene became completely, or at a high frequency, cytoplasmically petite. The subunits of bovine F1 were expressed individually in the yeast strains with the corresponding null mutations and targeted to the mitochondrion using a yeast mitochondrial leader peptide. Expression of the bovine alpha-, beta-, gamma-, and epsilon-, but not the delta-, subunit complemented the corresponding null mutations in yeast correcting the corresponding negative phenotypes. These results indicate that yeast is able to import, assemble subunits of bovine F1-ATPase in mitochondria and form a functional chimeric yeast/bovine enzyme complex.


Ventral Pallidum Is the Primary Target for Accumbens D1 Projections Driving Cocaine Seeking.
Outputs from the nucleus accumbens (NAc) include projections to the ventral pallidum and the ventral tegmental area and subtantia nigra in the ventral mesencephalon. The medium spiny neurons (MSN) that give rise to these pathways are GABAergic and consist of two populations of equal number that are segregated by differentially expressed proteins, including D1- and D2-dopamine receptors. Afferents to the ventral pallidum arise from both D1- and D2-MSNs, whereas the ventral mesencephalon is selectively innervated by D1-MSN. To determine the extent of collateralization of D1-MSN to these axon terminal fields we used retrograde labeling in transgenic mice expressing tdTomato selectively in D1-MSN, and found that a large majority of D1-MSN in either the shell or core subcompartments of the accumbens collateralized to both output structures. Approximately 70% of D1-MSNs projecting to the ventral pallidum collateralized to the ventral mesencephalon, whereas >90% of mesencephalic D1-MSN afferents collateralized to the ventral pallidum. In contrast, <10% of dorsal striatal D1-MSNs collateralized to both the globus pallidus and ventral mesencephalon. D1-MSN activation is required for conditioned cues to induce cocaine seeking. To determine which D1-MSN projection mediates cued cocaine seeking, we selectively transfected D1-MSNs in transgenic rats with an inhibitory Gi-coupled DREADD. Activation of the transfected Gi-DREADD with clozapine-N-oxide administered into the ventral pallidum, but not into the ventral mesencephalon, blocked cue-induced cocaine seeking. These data show that, although accumbens D1-MSNs largely collateralize to both the ventral pallidum and ventral mesencephalon, only D1-MSN innervation of the ventral pallidum is necessary for cue-induced cocaine seeking.SIGNIFICANCE STATEMENT Activity in D1 dopamine receptor-expressing neurons in the NAc is required for rodents to respond to cocaine-conditioned cues and relapse to drug seeking behaviors. The D1-expressing neurons project to both the ventral pallidum and ventral mesencephalon, and we found that a majority of the neurons that innervate the ventral pallidum also collateralize to the ventral mesencephalon. However, despite innervating both structures, only D1 innervation of the ventral pallidum mediates cue-induced cocaine seeking.


Comparison of Two Estrogen Chemically Activated Luciferase Expression Cell Bioassays to Liquid Chromatography-Mass Spectrometry for Quantifying Estrone in Water Samples.
Chemically activated luciferase expression (CALUX) cell bioassays are popular tools for assessing endocrine activity of chemicals such as certain environmental contaminants. Although activity equivalents can be obtained from CALUX analysis, directly comparing these equivalents to those obtained from analytical chemistry methods can be problematic because of the complexity of endocrine active pathways. We explored the suitability of two estrogen CALUX bioassays (the Organisation for Economic Co-operation and Development-approved VM7Luc4E2 cell bioassay and the VM7LucERβc9 cell bioassay) for quantitation of estrogen. Quadrupole-time of flight ultraperformance liquid chromatography-mass spectrometry (LC/MS) was selected as a comparative method. Regression analysis of measured estrone (E1) calibration samples showed all three methods to be highly predictive of nominal concentrations (p ≤ 7.5 × 10-51 ). Extracts of water sampled from laboratory dilutor tanks containing E1 at 0, 20, and 200 ng/L alone and in combination with atrazine were selected to test the quantitative capabilities of the CALUX assays. Process controls (0 and 100 ng E1/L) and a separate E1 standard (10 ng/ml, used to prepare the E1 process control) were also tested. Levels of E1 determined by LC/MS analysis and bioanalytical equivalents (ng E1/L) determined by CALUX analyses were comparable except in certain instances where the samples required dilution prior to CALUX analyses (e.g., the E1 process control and E1 standard). In those instances, measurements by CALUX were slightly but significantly decreased relative to LC/MS. Atrazine had no effect on the ability of either LC/MS or the CALUX bioassays to quantify E1. The present study illustrates the CALUX bioassays as successful in quantifying an estrogen in simple water samples and further characterizes their utility for screening. Environ Toxicol Chem 2023;42:333-339. Published 2022. This article is a U.S. Government work and is in the public domain in the USA.


[ENT hemangiomas - a new method for their treatment].
The objective of the present study was to address intricate problems of the treatment of ENT hemangiomas. Fibro-Vein preparation (FVP) was used for the first time for the treatment of the patients presenting with the vascular tumour in ENT. The clinical application of FVP was preceded by an experiment designed to determine the minimal dose needed to induce tissue sclerosis and adhesion. Monotherapy with FVP was given to 76 patients having the ENT hemangioma. The results of the treatment give reason to state that the use of Fibro-Vein for the treatment of the patients presenting with ENT hemangioma is an effective and sparing treatment modality.


The BHLF1 Locus of Epstein-Barr Virus Contributes to Viral Latency and B-Cell Immortalization.
The Epstein-Barr virus (EBV) BHLF1 gene encodes an abundant linear and several circular RNAs believed to perform noncoding functions during virus replication, although an open reading frame (ORF) is retained among an unknown percentage of EBV isolates. Evidence suggests that BHLF1 is also transcribed during latent infection, which prompted us to investigate the contribution of this locus to latency. Analysis of transcripts transiting BHLF1 revealed that its transcription is widespread among B-cell lines supporting the latency I or III program of EBV protein expression and is more complex than originally presumed. EBV-negative Burkitt lymphoma cell lines infected with either wild-type or two different BHLF1 mutant EBVs were initially indistinguishable in supporting latency III. However, cells infected with BHLF1- virus ultimately transitioned to the more restrictive latency I program, whereas cells infected with wild-type virus either sustained latency III or transitioned more slowly to latency I. Upon infection of primary B cells, which require latency III for growth in vitro, both BHLF1- viruses exhibited variably reduced immortalization potential relative to the wild-type virus. Finally, in transfection experiments, efficient protein expression from an intact BHLF1 ORF required the EBV posttranscriptional regulator protein SM, whose expression is limited to the replicative cycle. Thus, one way in which BHLF1 may contribute to latency is through a mechanism, possibly mediated or regulated by a long noncoding RNA, that supports latency III critical for the establishment of EBV latency and lifelong persistence within its host, whereas any retained protein-dependent function of BHLF1 may be restricted to the replication cycle.IMPORTANCE Epstein-Barr virus (EBV) has significant oncogenic potential that is linked to its latent infection of B lymphocytes, during which virus replication is not supported. The establishment of latent infection, which is lifelong and can precede tumor development by years, requires the concerted actions of nearly a dozen EBV proteins and numerous small non-protein-coding RNAs. Elucidating how these EBV products contribute to latency is crucial for understanding EBV's role in specific malignancies and, ultimately, for clinical intervention. Historically, EBV genes that contribute to virus replication have been excluded from consideration of a role in latency, primarily because of the general incompatibility between virus production and cell survival. However, here, we provide evidence that the genetic locus containing one such gene, BHLF1, indeed contributes to key aspects of EBV latency, including its ability to promote the continuous growth of B lymphocytes, thus providing significant new insight into EBV biology and oncogenic potential.


Evaluation of the efficacy of humic acids to counteract the toxic effects of aflatoxin B1 in turkey poults.
This study aims to evaluate the efficacy of humic acid (HA) from worm compost as an adsorbent for aflatoxin B1 (AFB1) in turkey poults. The experiment involved the inclusion of 0.25% (w/w) HA in the diet of turkey poults consuming aflatoxin-contaminated feed (250 ng AFB1/g). A total of 350 1-day-old female Nicholas-700 turkey poults were randomly allocated to five equal groups: negative control (basal diet); positive control (basal diet + 250 ng AFB1/g; HA (basal diet + 0.25% HA); HA + AFB1 (basal diet + HA + 250 ng AFB1/g); and zeolite + AFB1 (basal diet + 0.25% zeolite + 250 ng AFB1/g). Each group had seven replicates of 10 poults (n = 70). The impact of HA addition was evaluated in terms of performance parameters, relative organ weights, liver histological lesions, and serum biochemical and hematological constituents. In general, the addition of HA improved body weight (BW), body weight gain (BWG), and feed conversion rate (FCR). Furthermore, HA effectively mitigated the toxic effects caused by AFB1 in the majority of the analyzed variables. The results indicated that HA effectively counteracted the AFB1-induced toxic effects in turkey poults. Based on these findings, it can be concluded that HA is capable of removing AFB1 from the contaminated diet.


Lactate Metabolism, Signaling, and Function in Brain Development, Synaptic Plasticity, Angiogenesis, and Neurodegenerative Diseases.
Neural tissue requires a great metabolic demand despite negligible intrinsic energy stores. As a result, the central nervous system (CNS) depends upon a continuous influx of metabolic substrates from the blood. Disruption of this process can lead to impairment of neurological functions, loss of consciousness, and coma within minutes. Intricate neurovascular networks permit both spatially and temporally appropriate metabolic substrate delivery. Lactate is the end product of anaerobic or aerobic glycolysis, converted from pyruvate by lactate dehydrogenase-5 (LDH-5). Although abundant in the brain, it was traditionally considered a byproduct or waste of glycolysis. However, recent evidence indicates lactate may be an important energy source as well as a metabolic signaling molecule for the brain and astrocytes-the most abundant glial cell-playing a crucial role in energy delivery, storage, production, and utilization. The astrocyte-neuron lactate-shuttle hypothesis states that lactate, once released into the extracellular space by astrocytes, can be up-taken and metabolized by neurons. This review focuses on this hypothesis, highlighting lactate's emerging role in the brain, with particular emphasis on its role during development, synaptic plasticity, angiogenesis, and disease.


Evidence of thrombotic microangiopathy in children with SARS-CoV-2 across the spectrum of clinical presentations.
Most children with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection have mild or minimal disease, with a small proportion developing severe disease or multisystem inflammatory syndrome in children (MIS-C). Complement-mediated thrombotic microangiopathy (TMA) has been associated with SARS-CoV-2 infection in adults but has not been studied in the pediatric population. We hypothesized that complement activation plays an important role in SARS-CoV-2 infection in children and sought to understand if TMA was present in these patients. We enrolled 50 hospitalized pediatric patients with acute SARS-CoV-2 infection (n = 21, minimal coronavirus disease 2019 [COVID-19]; n = 11, severe COVID-19) or MIS-C (n = 18). As a biomarker of complement activation and TMA, soluble C5b9 (sC5b9, normal 247 ng/mL) was measured in plasma, and elevations were found in patients with minimal disease (median, 392 ng/mL; interquartile range [IQR], 244-622 ng/mL), severe disease (median, 646 ng/mL; IQR, 203-728 ng/mL), and MIS-C (median, 630 ng/mL; IQR, 359-932 ng/mL) compared with 26 healthy control subjects (median, 57 ng/mL; IQR, 9-163 ng/mL; P < .001). Higher sC5b9 levels were associated with higher serum creatinine (P = .01) but not age. Of the 19 patients for whom complete clinical criteria were available, 17 (89%) met criteria for TMA. A high proportion of tested children with SARS-CoV-2 infection had evidence of complement activation and met clinical and diagnostic criteria for TMA. Future studies are needed to determine if hospitalized children with SARS-CoV-2 should be screened for TMA, if TMA-directed management is helpful, and if there are any short- or long-term clinical consequences of complement activation and endothelial damage in children with COVID-19 or MIS-C.


Human papillomavirus vaccination in the European Union/European Economic Area and globally: a moral dilemma.
While many European Union/European Economic Area (EU/EEA) countries recently expanded human papillomavirus (HPV) vaccination to boys, HPV vaccine supply is currently limited for girls in low- and middle-income countries (LMIC) that are severely affected by HPV.Globally, about 50% of countries have introduced HPV vaccination. Some LMIC with high burden of cervical cancer have not yet introduced HPV vaccination, or are reaching suboptimal vaccination coverage. While WHO issued a call for cervical cancer elimination in 2018, a global shortage of HPV vaccines is currently predicted to last at least until 2024.We reviewed national policies of EU/EEA countries and recommendations of the World Health Organization (WHO) Strategic Advisory Group of Experts on immunisation to discuss current challenges and dose-sparing options. Several EU/EEA countries have extended HPV vaccination to boys and the European Cancer Organisation has issued a resolution for elimination of all HPV-associated cancers in both sexes. The European Centre for Disease Prevention and Control concluded in its 2020 guidance that cost-effectiveness of extending routine vaccination to boys depends on several context-specific factors. The extension of HPV vaccination to boys in EU/EEA countries may affect global availability of vaccines. Temporary dose-sparing options could be considered during the COVID-19 post-pandemic period.


Visual Analytics for Topic Model Optimization based on User-Steerable Speculative Execution.
To effectively assess the potential consequences of human interventions in model-driven analytics systems, we establish the concept of speculative execution as a visual analytics paradigm for creating user-steerable preview mechanisms. This paper presents an explainable, mixed-initiative topic modeling framework that integrates speculative execution into the algorithmic decisionmaking process. Our approach visualizes the model-space of our novel incremental hierarchical topic modeling algorithm, unveiling its inner-workings. We support the active incorporation of the user's domain knowledge in every step through explicit model manipulation interactions. In addition, users can initialize the model with expected topic seeds, the backbone priors. For a more targeted optimization, the modeling process automatically triggers a speculative execution of various optimization strategies, and requests feedback whenever the measured model quality deteriorates. Users compare the proposed optimizations to the current model state and preview their effect on the next model iterations, before applying one of them. This supervised human-in-the-loop process targets maximum improvement for minimum feedback and has proven to be effective in three independent studies that confirm topic model quality improvements.


Clutch size in the tropical scincid lizard Emoia sanfordi, a species endemic to the Vanuatu Archipelago.
The majority of species in the scincid genus Emoia (Squamata: Scincidae) have a fixed clutch size of two eggs per clutch and produce between two and four clutches per year. One lineage within Emoia, the Emoia samoensis species group, consists of 13 species occurring in Melanesia and the islands of the southwestern Pacific Ocean, and exhibits variation in clutch size, with previously reported clutch sizes of two to five eggs. Little is known about reproduction in several members of this lineage including Emoia sanfordi, a large-bodied lizard endemic to the archipelago of Vanuatu in the South Pacific. We analyzed reproduction and clutch size in E. sanfordi females and discovered that there is a substantial amount of intraspecific variation, with clutch size ranging from two to seven eggs, with a modal clutch size of five eggs. Females were reproductively active throughout the study period of June through October and appear to be laying multiple clutches. The variation in clutch size seen in E. sanfordi is congruent with the variation previously reported within other closely related species.


How adolescents with ASD process social information in complex scenes. combining evidence from eye movements and verbal descriptions.
We investigated attention, encoding and processing of social aspects of complex photographic scenes. Twenty-four high-functioning adolescents (aged 11-16) with ASD and 24 typically developing matched control participants viewed and then described a series of scenes, each containing a person. Analyses of eye movements and verbal descriptions provided converging evidence that both groups displayed general interest in the person in each scene but the salience of the person was reduced for the ASD participants. Nevertheless, the verbal descriptions revealed that participants with ASD frequently processed the observed person's emotion or mental state without prompting. They also often mentioned eye-gaze direction, and there was evidence from eye movements and verbal descriptions that gaze was followed accurately. The combination of evidence from eye movements and verbal descriptions provides a rich insight into the way stimuli are processed overall. The merits of using these methods within the same paradigm are discussed.


Gastric potential difference measurements. The gastric mucosal integrity and function studied with a new method for measurement of the electric potential difference across the stomach wall.
PD--the electric potential difference across the gastric mucosa--is a variable used to describe the gastric mucosal integrity and function. A new, reliable, and easily applied method for gastric PD measurements corrected for the disturbing liquid junction potentials between gastric juice and the PD measuring probe is presented. PD is measured with the gastric lumen negative, and a numeric reduction in PD is used as an expression of an injured mucosal condition. A reduced gastric PD is found along with a reduced gastric mucosal blood flow after intravenous indomethacin in anesthetized dogs. Increasing the FFA/albumin ratios in mini-pigs causes vasoconstriction and PD reduction. Short hypoxia and selective gastric ischemia cause a reversible PD reduction and no morphologic changes in anesthetized dogs, but ischemia for 1 h causes more permanent changes in PD, pH, and morphology. This damage can be reduced by allopurinol pretreatment, possibly due to the inhibition of oxygen-derived free radical formation. Gastric PD and pH were measured in volunteers and duodenal ulcer patients during Stroop's color word conflict test, in which mental stress causes sympathetic activation. A PD reduction and a pH increase were found along with stress induction, thereby indicating an influence of mental stress on stomach mucosal function. It is concluded that gastric PD measurement may be useful in ulcer pathogenetic research, and a sufficient gastric mucosal blood flow is stressed as being important for the mucosal defense.


The effect of cortisol on the synthesis of rat plasma albumin, fibrinogen and transferrin.
A decrease of absolute synthesis of albumin, no change in that of fibrinogen and an increased fractional synthesis of transferrin were observed 3h after intraperitoneal administration of a pharmacological dose of 5 mg of cortisol to 220g rats in the post-absorptive state and previously kept on a diet with 40% protein. The concentration in liver of total free amino acids was practically unchanged at this time. Intraperitoneal administration of a mixture of amino acids with the cortisol raised this concentration and was accompanied by an almost complete de-repression of the synthesis of albumin, with no real effect on that of fibrinogen. In considerable contrast, in rats studied at 24h after intraperitoneal administration of cortisol, and who had been fed once in the interim (but who had received no amino acids intraperitoneally), there was a marked increase in the absolute synthesis of albumin and fibrinogen, with an increase in fractional synthesis that was less proportionately but still very significant and which included transferrin. The amino acid concentrations had risen above the supplemented values at 3h but not as much proportionately as the fractional synthesis rates, and of course not as much as the absolute synthesis rates, of albumin and fibrinogen. These time-dependent effects of cortisol suggest to us that our studies resolve the apparently conflicting results of the effect of cortisol on the synthesis of albumin reported by others.


Clustering Treatment Outcomes in Women with Gambling Disorder.
The rising prevalence of gambling disorder (GD) among women has awakened considerable interest in the study of therapeutic outcomes in females. This study aimed to explore profiles of women seeking treatment for GD based on a set of indicators including sociodemographic features, personality traits, clinical state at baseline, and cognitive behavioral therapy (CBT) outcomes. Two-step clustering, an agglomerative hierarchical classification system, was applied to a sample of n = 163 women of ages ranging from 20 to 73 years-old, consecutively attended to by a clinical unit specialized in the treatment of G. Three mutually exclusive clusters were identified. Cluster C1 (n = 67, 41.1%) included the highest proportion of married, occupationally active patients within the highest social status index. This cluster was characterized by medium GD severity levels, the best psychopathological functioning, and the highest mean in the self-directedness trait. C1 registered 0% dropouts and only 14.9% relapse. Cluster C2 (n = 63; 38.7%) was characterized by the lowest GD severity, medium scores for psychopathological measures and a high risk of dropout during CBT. Cluster C3 (n = 33; 20.2%) registered the highest GD severity, the worst psychopathological state, the lowest self-directedness level and the highest harm-avoidance level, as well as the highest risk of relapse. These results provide new evidence regarding the heterogeneity of women diagnosed with GD and treated with CBT, based on the profile at pre- and post-treatment. Person-centered treatments should include specific strategies aimed at increasing self-esteem, emotional regulation capacities and self-control of GD women.


The contribution of diet, physical activity and sedentary behaviour to body mass index in women with and without polycystic ovary syndrome.
What is the contribution of diet, physical activity and sedentary behaviour to body mass index (BMI) in women with and without polycystic ovary syndrome (PCOS)? PCOS status, higher energy intake and glycaemic index and lower physical activity were independently associated with BMI. Obesity worsens the clinical features of PCOS and women with PCOS have an elevated prevalence of overweight and obesity. It is not known whether there is a contribution of lifestyle factors such as dietary intake, physical activity or sedentary behaviour to the elevated prevalence of obesity in PCOS. This study is a population-based observational study with data currently collected at 13 year follow-up. The study commenced in 1996. For this analysis, data are analysed at one time point corresponding to the Survey 5 of the cohort in 2009. At this time 8200 participants remained (58% retention of baseline participants) of which 7466 replied to the questionnaire; 409 self-reported a diagnosis of PCOS and 7057 no diagnosis of PCOS. Australian women born in 1973-1978 from the Australian Longitudinal Study on Women's Health. Mean BMI was higher in women with PCOS compared with non-PCOS (29.3 ± 7.5 versus 25.6 ± 5.8 kg/m(2), P < 0.001). Women with PCOS reported a better dietary intake (elevated diet quality and micronutrient intake and lower saturated fat and glycaemic index intake) but increased energy intake, increased sitting time and no differences in total physical activity compared with non-PCOS. PCOS status, higher energy intake and glycaemic index and lower physical activity, as well as age, smoking, alcohol intake, occupation, education and country of birth, were independently associated with BMI. The weaknesses of this study include the self-reported diagnosis of PCOS, and the women not reporting PCOS not having their control status clinically verified which is likely to underrepresent the PCOS population. We are also unable to determine if lifestyle behaviours contributed to the PCOS diagnosis or were altered in response to diagnosis. The strengths of this study include the community-based nature of the sample which minimizes selection bias to include women with a variety of clinical presentations. These results are therefore generalizable to a broader population than the majority of research in PCOS examining this research question which are performed in clinic-based populations. This study is in agreement with the literature that PCOS is independently associated with elevated BMI. We provide new insights that diet quality is subtly improved but that sedentary behaviour is elevated in PCOS and that PCOS status, higher energy intake and glycaemic index and lower physical activity are independently associated with BMI. L.J.M. was supported by a South Australian Cardiovascular Research Development Program (SACVRDP) Fellowship (AC11S374); a program collaboratively funded by the National Heart Foundation of Australia, the South Australian Department of Health and the South Australian Health and Medical Research Institute, S.A.M. was funded by an Australian Research Council Future Fellowship (FT100100581), S.Z. was funded by a Heart Foundation Career Development Fellowship (ID CR10S5330) and H.J.T. was funded by an NHMRC fellowship (ID 545888). None of the authors has any conflict of interest to declare. Not applicable.


Somatic genomic variations in extra-embryonic tissues.
In the mature chorion, one of the membranes that exist during pregnancy between the developing fetus and mother, human placental cells form highly specialized tissues composed of mesenchyme and floating or anchoring villi. Using fluorescence in situ hybridization, we found that human invasive cytotrophoblasts isolated from anchoring villi or the uterine wall had gained individual chromosomes; however, chromosome losses were detected infrequently. With chromosomes gained in what appeared to be a chromosome-specific manner, more than half of the invasive cytotrophoblasts in normal pregnancies were found to be hyperdiploid. Interestingly, the rates of hyperdiploid cells depended not only on gestational age, but were strongly associated with the extraembryonic compartment at the fetal-maternal interface from which they were isolated. Since hyperdiploid cells showed drastically reduced DNA replication as measured by bromodeoxyuridine incorporation, we conclude that aneuploidy is a part of the normal process of placentation potentially limiting the proliferative capabilities of invasive cytotrophoblasts. Thus, under the special circumstances of human reproduction, somatic genomic variations may exert a beneficial, anti-neoplastic effect on the organism.


Rapid LA-REIMS and comprehensive UHPLC-HRMS for metabolic phenotyping of feces.
Ambient ionization-based techniques hold great potential for rapid point-of-care applicable metabolic fingerprinting of tissue and fluids. Hereby, feces represents a unique biospecimen as it integrates the complex interactions between the diet, gut microbiome and host, and is therefore ideally suited to study the involvement of the diet-gut microbiome axis in metabolic diseases and their treatments at a molecular level. We present a new method for rapid (<10 s) metabolic fingerprinting of feces, i.e. laser-assisted rapid evaporative ionization mass spectrometry (LA-REIMS) with an Nd:YAG laser (2940 nm) and quadrupole Time-of-Flight mass spectrometer as main components. The LA-REIMS method was implemented on mimicked crude feces samples from individuals that were assigned a state of type 2 diabetes or euglycaemia. Based on the generated fingerprints, enclosing 4923 feature ions, significant segregation according to disease classification was achieved through orthogonal partial least squares discriminant analysis (Q2(Y) of 0.734 and p-value of 1.93e-17) and endorsed by a general classification accuracy of 90.5%. A comparison between the discriminative performance of the novel LA-REIMS and our established ultra-high performance liquid-chromatography high-resolution MS (UHPLC-HRMS) metabolomics and lipidomics methodologies for fingerprinting of stool was performed. Based on the supervised modelling results upon UHPLC-HRMS (Q2(Y) ≥ 0.655 and p-value ≤ 4.11 e-5), equivalent or better discriminative performance of LA-REIMS fingerprinting was concluded. Eventually, comprehensive UHPLC-HRMS was employed to assess metabolic alterations as observed for the defined classes, whereby metformin treatment of the type 2 diabetes patients was considered a relevant study factor to acquire new mechanistic insights. More specifically, ten metabolization products of metformin were identified, with (hydroxylated) triazepinone and metformin-cholesterol reported for the first time in vivo.In conclusion, LA-REIMS was established as an expedient strategy for rapid metabolic fingerprinting of feces, whereby potential implementations may relate, but are not limited to differential diagnosis and treatment efficacy evaluation of metabolic diseases. Yet, LC-HRMS remains essential for in-depth biological interpretation.


Liver stiffness and fibrosis-4 alone better predict liver events compared with aspartate aminotransferase to platelet ratio index in a cohort of human immunodeficiency virus and hepatitis C virus co-infected patients from ANRS CO13 HEPAVIH cohort.
HIV/hepatitis C virus (HCV) co-infection leads to major complications, and noninvasive markers developed to stage liver fibrosis could be used as prognostic markers. We aimed to compare the performances of liver stiffness (LS), fibrosis-4 (FIB-4), and aspartate aminotransferase to platelet ratio index (APRI) to predict liver-related events in HIV/HCV co-infected patients. HIV/HCV co-infected patients from the ANRS CO13 HEPAVIH cohort were included if they had LS, FIB-4, and APRI measurements done in a window of 3 months. Primary outcome was the time between inclusion and occurrence of a liver-related event. Univariable and multivariable Fine and Gray models were performed. Predictive performances were compared by the area under the receiver operating characteristic (AUROC) differences after correction of optimistic by bootstrap samples. Best cutoffs to predict liver-related events were estimated by sensitivity and specificity maximization. A total of 998 patients were included. Overall, 70.7% were men. Their median age was 46.8 years. According to LS value, 204 (20.4%) patients had cirrhosis. Overall, 39 patients experienced at least one liver-related event. In univariable analysis, LS AUROC curve was significantly superior to FIB-4 and APRI AUROC curves, being 87.9, 78.2, and 75.0%, respectively. After adjustment on age, CD4 levels, and insulin resistance, no differences were observed. The best cutoffs to identify patients at low or high risk of liver-related events were below 8.5, 1.00, and 0.35 and above 16.5, 4.00, and 1.75 for LS, FIB-4, and APRI, respectively. To predict HCV-related events, APRI had lower performance than LS and FIB-4. FIB-4 is as good as LS to predict HCV-related events, suggesting that it can be used for the management of HIV/HCV co-infected patients and replace LS.


Phytotoxic and nematicide evaluation of Croton ehrenbergii (Euphorbiaceae).
Within its natural habitat, Croton ehrenbergii exhibits an innate defense mechanism that is not seen in other plants; it grows unharmed amidst predators and nature, while other species perish. In light of its capacity for defense, C. ehrenbergii was evaluated to better understand the scope of its phytotoxic and nematicide properties. To assess this, fractions obtained by bipartition of a hydro-alcoholic extract, and l-quebrachitol, the main constituent of C. ehrenbergii, were evaluated on Lactuca sativa (dicotyledon) and Lolium perenne (monocotyledon) seeds. Additionally, bipartition fractions and l-quebrachitol were evaluated on a population of Meloidogyne incognita for their nematicide activity. From this phytochemical research, l-quebrachitol (1), phenylalanine (2), trans-4-hydroxy-N-methylproline (3) and the flavonoids: kaempferol (4), tiliroside (5), nicotiflorine (6) and rutin (7) were identified by spectroscopic analysis. Both methanol and hexane fractions from aerial parts of the plant inhibited the germination and elongation of roots and stems in L. sativa, but not in Lolium perenne, showing that these fractions mostly inhibit the dicotyledon species used in this research. l-Quebrachitol showed slightly higher seed germination inhibition for Lolium perenne in comparison with L. sativa. Three of the fractions evaluated showed nematicide activity against Meloidogyne incognita larvae (J2) at the 48 h benchmark, compared with carbofuran. l-Quebrachitol shows higher mortality after 48 h exposure at a lower concentration than carbofuran. A variety of compounds were isolated from this research, some were common within the Croton genus (4-7), whereas others were not (1-3). This is the first phytochemical, phytotoxic and nematicide report on C. ehrenbergii. Methanol and hexane fractions from the aerial parts of C. ehrenbergii and l-quebrachitol could be used as alternative natural herbicides, predominantly against dicotyledon weed seeds. All fractions had a good mortality percentage against J2 Meloidogyne incognita larvae. © 2019 Society of Chemical Industry.


Seasonal and Annual Change in Physiological Ocular Growth of 7- to 11-Year-Old Norwegian Children.
To investigate seasonal and annual change in physiological eye growth in Norwegian school children. Measurements of ocular biometry, non-cycloplegic spherical equivalent autorefraction (SER), and choroidal thickness (ChT) were obtained for 92 children (44 females) aged 7 to 11 years at four time points over a year (November 2019-November 2020). Seasons (3- and 5-month intervals) were classified as winter (November-January), winter-spring (January-June), and summer-autumn (June-November). Cycloplegic SER was obtained in January and used to group children. The seasonal and annual changes were tested with a linear mixed-effects model (P values were adjusted for multiple comparisons). All the children experienced annual ocular growth, irrespective of SER, but less so during the summer-autumn. The baseline SER was lower (P < 0.001), axial length (AL) was longer (P < 0.038), and choroids were thicker in 10- to 11-year-old than 7- to 8-year-old mild hyperopes (P = 0.002). Assuming mild hyperopes (n = 65) experience only physiological eye growth, modeling revealed seasonal and annual increases in AL across sex and age (P < 0.018), with less change during the summer-autumn than winter-spring. The 7- to 8-year-olds had a larger decrease annually and over winter-spring in SER (P ≤ 0.036) and in ChT over winter-spring than the 10- to 11-year-olds (P = 0.006). There were significant seasonal and annual changes in AL in children who had physiological eye growth irrespective of age within this cohort. Annual changes in SER and seasonal choroidal thinning were only observed in 7- to 8-year-old children. This indicates continued emmetropization in 7- to 8-year-olds and a transition to maintaining emmetropia in 10- to 11-year-olds.


Clinical care for obesity: A preliminary survey of sixty-eight countries.
Obesity is a chronic relapsing condition affecting a rapidly increasing number of people worldwide. The United Nations has stated that universal health coverage is an essential element of the globally-agreed sustainable development goals. This article provides a preliminary report of a survey of relevant health professionals and other interest groups on the readiness of health systems to provide obesity treatment services. Interviews and questionnaires were completed by 274 respondents from a total of 68 low, middle and high income countries. Respondents in the majority of countries stated that there were professional guidelines for obesity treatment, but that there was a lack of adequate services, especially in lower income countries, and in rural areas of most countries. Lack of treatment was attributed to a broad range of issues including: no clear care pathways from primary care to secondary services; absent or limited secondary services in some regions; lack of trained multi-disciplinary support professionals; potentially high costs to patients; long waiting times for surgery; and stigma experienced by patients within the health care services. Defining obesity as a disease may help to overcome stigma and may also help to secure better funding streams for treatment services. However, the survey found that few countries were ready to accept this definition. Furthermore, until countries fully adopt and implement obesity prevention policies the need for treatment will continue to rise while the necessary conditions for treatment will remain inadequate.


Living Unnoticed: Cognitive Impairment in Older People with Multimorbidity.
To investigate the correlation between MMSE ≤ 23 and the presence of a diagnosis of dementia in the medical record in a population with multimorbidity. This cross-sectional study was part of the Ambulatory Geriatric Assessment - a Frailty Intervention Trial (AGe-FIT; N = 382). Participants were community dwelling, aged ≥ 75 years, had received inpatient hospital care at least three times during the past 12 months, and had three or more concomitant diagnoses according to the International Classification of Diseases, 10th revision. The Mini Mental State Examination (MMSE) was administered at baseline. Medical records of participants with MMSE scores < 24 were examined for the presence of dementia diagnoses and two years ahead. Fifty-three (16%) of 337 participants with a measure of MMSE had a MMSE scores < 24. Six of these 53 (11%) participants had diagnoses of dementia (vascular dementia, n = 4; unspecified dementia, n = 1; Alzheimers disease, n = 1) according to medical records; 89% did not. A MMSE-score < 24 is not well correlated to a diagnosis of dementia in the medical record in a population of elderly with multimorbidity. This could imply that cognitive decline and the diagnosis of dementia remain undetected in older people with multimorbidity. Proactive care of older people with multimorbidity should focus on cognitive decline to detect cognitive impairment and to provide necessary help and support to this very vulnerable group.


Clinical Study of Mitomycin C in Reducing Haze Formation After Ultraviolet A/Riboflavin Crosslinking for Keratoconus.
To evaluate the safety and efficacy of mitomycin C (MMC) in haze formation after ultraviolet A/riboflavin corneal crosslinking (CXL) for progressive keratoconus. A total of 60 keratoconic eyes of 48 patients were enrolled in this prospective comparative study after obtaining informed consent. In the CXL group, standard corneal CXL was performed, whereas in the CXL+MMC group, 0.02% MMC was used for 30 s soon after CXL. Comprehensive ophthalmologic examinations were performed on all patients before surgery and at 1, 3, 6, and 12 months after surgery. The epithelium recovered within 3 to 4 days after CXL, and the healing time was comparable in the two groups. There was no significant endothelial cell density loss after CXL in both groups. Eyes in both groups showed improvement of uncorrected distance visual acuity (Snellen) and best-corrected visual acuity (Snellen; P<0.05), and there was a decrease in K-max, cylinder degree, and central corneal thickness (CCT) (P<0.05). There was no significant statistical difference between the groups regarding postoperative K-max reduction, refraction, and CCT (P>0.05). Corneal haze scores were significantly higher in the CXL group at 1 and 3 months after CXL (P=0.012 and P=0.028, respectively), but were similar to the MMC group at 6 and 12 months after surgery (P=0.329 and P=0.543, respectively). Prophylactic intraoperative use of 0.02% MMC can significantly reduce CXL-associated haze formation, especially in the early postoperative period, and no signs of weakening CXL efficacy were observed.


A QRS scoring system for assessing left ventricular function after myocardial infarction.
A QRS scoring system for estimating the size of a myocardial infarct was evaluated in 55 patients who did not have left ventricular hypertrophy or conduction abnormalities. Serial 12-lead surface electrocardiograms were scored according to a 29-point system based on the duration of Q and R waves and on the ratios of R-to-Q amplitude and R-to-S amplitude. The scores were proportional to the severity of wall-motion abnormalities, which was determined by radionuclide blood-pool scanning and which correlated inversely with the radionuclide-determined left ventricular ejection fraction (LVEF). A score less than 3 was 93 per cent sensitive and 88 per cent specific for both severe regional dyssynergy and major depression of the global LVEF. The following equation was used to estimate the LVEF from the QRS score: LVEF (%) = 60 - (3 x QRS score). After acute myocardial infarction, an electrocardiogram can provide important indirect quantitative information about left ventricular function.


Improved Lasso for genomic selection.
Empirical experience with genomic selection in dairy cattle suggests that the distribution of the effects of single nucleotide polymorphisms (SNPs) might be far from normality for some traits. An alternative, avoiding the use of arbitrary prior information, is the Bayesian Lasso (BL). Regular BL uses a common variance parameter for residual and SNP effects (BL1Var). We propose here a BL with different residual and SNP effect variances (BL2Var), equivalent to the original Lasso formulation. The λ parameter in Lasso is related to genetic variation in the population. We also suggest precomputing individual variances of SNP effects by BL2Var, to be later used in a linear mixed model (HetVar-GBLUP). Models were tested in a cross-validation design including 1756 Holstein and 678 Montbéliarde French bulls, with 1216 and 451 bulls used as training data; 51 325 and 49 625 polymorphic SNP were used. Milk production traits were tested. Other methods tested included linear mixed models using variances inferred from pedigree estimates or integrated out from the data. Estimates of genetic variation in the population were close to pedigree estimates in BL2Var but not in BL1Var. BL1Var shrank breeding values too little because of the common variance. BL2Var was the most accurate method for prediction and accommodated well major genes, in particular for fat percentage. BL1Var was the least accurate. HetVar-GBLUP was almost as accurate as BL2Var and allows for simple computations and extensions.


Formulation development and evaluation of Diltiazem HCl sustained release matrix tablets using HPMC K4M and K100M.
The aim of this study was to develop a sustained release hydrophilic matrix tablet of Diltiazem HCl and evaluates the effect of formulation variables (e.g. lubricant, binder, polymer content and viscosity grades of HPMC) on drug release. Twelve different formulations (F1-F12) were prepared by direct compression. The results of the physical parameters and assay were found to be within the acceptable range. Rate of drug release was found to be slow as the fraction of the polymer was increased from 20-50%. The drug release rate from tablets containing K4M was effectively controlled by increasing the talc concentration, whereas the burst effect was reduced by increasing binder content. The drug release was higher with K4M as compare to K100M. Model-dependent and independent methods were used for data analysis and the best results were observed for K4M in Higuchi (R(2)=0.9903-0.9962) and K100M in Baker and Lonsdale (R(2)=0.9779-0.9941). The release mechanism of all formulations was non-Fickian. F7 (50% K4M, 2% talc, 10% Avicel PH101) and F11 (40% K100M) were very close to targeted release profile. F12 (50% K100M) exhibited highest degree of swelling and lowest erosion. The f1 and f2 test were performed taking F11 as a reference formulation.


Electroencephalographic evaluation of cerebral hyperperfusion syndrome following superficial temporal artery-middle cerebral artery anastomosis.
Low-flow bypass, such as superficial temporal artery-middle cerebral artery (STA-MCA) anastomosis, can result in cerebral hyperperfusion syndrome (CHS). The present study evaluated the pathophysiological conditions of CHS through the use of repeated electroencephalography (EEG). Among a total of 22 patients who underwent STA-MCA anastomosis over a course of 4 years, 3 patients were diagnosed with CHS based on clinical symptoms and neuroradiological examinations, including cerebral blood flow evaluation. Case 1 and Case 2 developed CHS on postoperative day 1, when EEG demonstrated focal slow waves on the frontal region of the operated side, indicating cortical dysfunction in these areas. Although prompt recovery of these EEG findings was noted with improvement of the clinical symptoms in Case 1, Case 2 developed an intracranial hemorrhage on postoperative day 5, when EEG clearly depicted persistent nonconvulsive status epilepticus (NCSE) after control of convulsive status epilepticus. In contrast, the clinical onset in Case 3 was delayed to postoperative day 6 and EEG revealed frequent ictal discharges in the operated hemisphere, although convulsive seizures were not apparent. Administration of anticonvulsants was performed after the diagnosis of NCSE, and complete recovery from CHS was achieved. Although the pathophysiology of CHS is cortical dysfunction, ictal hyperperfusion associated with NCSE could be included. The present findings emphasize the importance of repeated EEG examinations in the differential diagnosis of the various types of pathophysiological conditions of CHS.


[Retinoic acid inhibits tumor necrosis factor-alpha induced injury in human lung epithelial cells].
To study the effect of tumor necrosis factor-alpha (TNF-alpha) on the proliferation and apoptosis of type II lung alveolar epithelial cells and the regulation of retinoic acid (RA) to this effect. Human type II lung alveolar epithelial cells of the line A569 were cultured and divided into 2 groups: control group (cultured in culture fluid only), TNF-group (cultured in the culture fluid with TNF-alpha 10 micromol/L for 24 or 46 h respectively), RA group (cultured in culture fluid without RA 1 microg/L), and TNF-alpha plus RA group (cultured in culture fluid with TNF-alpha 10 micromol/L + RA 1 microg/L). MTT method was used to test the proliferation of the A549 cells. The cell apoptosis was detected by flow cytometric assay. The proliferation rates of A549 cells treated with TNF-alpha of the concentrations of 0, 0.1, 1, 5, and 10 microg/L were 95.0%, 90.0%, 79.6%, 72.4%, and 59.6% after 24 h (F = 18.04, P < 0.001), and were 93.2%, 82.7%, 61.5%, 50.3%, and 44.7% after 48 h (F = 40.61, P < 0.0001). The inhibition effect on the proliferation of A549 cells treated with 10 microg/L TNF-alpha for 24 h could be reversed, however, the inhibition effect on the proliferation of A549 cells treated with 10 microg/L TNF-a for 48h could not be reversed. RA alone did not significantly promote the proliferation of the A549 cells, but weakened the inhibitory effect of TNF-alpha on the proliferation of the A549 cells. The apoptotic rate of the A549 cells treated with TNF-alpha for 12 h, 24 h and 48 h respectively were 14.3% +/- 3.2%, 18.6% +/- 2.9%, and 43.4% +/- 3.5% respectively, all significantly higher than those of the control group (6.3% +/- 1.2%, 8.2% +/- 1.3%, and 26.1% +/- 2.5% respectively, all P < 0.01), and the apoptotic rate of the A549 cells treated with both TNF-alpha and RA for 12 h, 24 h, and 48h were 4.8% +/- 1.1%, 5.2% +/- 1.3%, and 16.4% +/- 2.3% respectively, all significantly lower than those of the TNF-alpha group (all P < 0.01), and the apoptotic rates of the A549 cells treated with both TNF-alpha and RA for 24 h and 48 h respectively were both significantly lower than those of the control group (both P < 0.05). RA relieves the injury of alveolar epithelial cells and protects the pulmonary surfactant by inhibiting the destruction of TNF-alpha to type II lung alveolar epithelial cells.


Critical role of 7,8-didemethyl-8-hydroxy-5-deazariboflavin for photoreactivation in Chlamydomonas reinhardtii.
DNA photolyases use two noncovalently bound chromophores to catalyze photoreactivation, the blue light-dependent repair of DNA that has been damaged by ultraviolet light. FAD is the catalytic chromophore for all photolyases and is essential for photoreactivation. The identity of the second chromophore is often 7,8-didemethyl-8-hydroxy-5-deazariboflavin (FO). Under standard light conditions, the second chromophore is considered nonessential for photoreactivation because DNA photolyase bound to only FAD is sufficient to catalyze the repair of UV-damaged DNA. phr1 is a photoreactivation-deficient strain of Chlamydomonas. In this work, the PHR1 gene of Chlamydomonas was cloned through molecular mapping and shown to encode a protein similar to known FO synthases. Additional results revealed that the phr1 strain was deficient in an FO-like molecule and that this deficiency, as well as the phr1 photoreactivation deficiency, could be rescued by transformation with DNA constructs containing the PHR1 gene. Furthermore, expression of a PHR1 cDNA in Escherichia coli produced a protein that generated a molecule with characteristics similar to FO. Together, these results indicate that the Chlamydomonas PHR1 gene encodes an FO synthase and that optimal photoreactivation in Chlamydomonas requires FO, a molecule known to serve as a second chromophore for DNA photolyases.


Evaluating the partial contribution of the P3 event-related potential elicited by auditory oddball stimuli during the Stroop task.
The cognitive load of a precisely timed task, such as the Stroop task, may be measured through the use of event-related potentials (ERPs). To determine the time at which cognitive load is at its peak, oddball tones may be applied at various times surrounding a cognitive task. However, we need to determine whether the simultaneous presentation of auditory and visual stimuli would mask a potential change in P3 in an ERP-producing task. If the contribution of the Stroop stimulus is too large, then Stroop ERP with oddball stimuli occurring at different timepoints may not be directly comparable across the various timepoints due to the contribution of the Stroop ERP. The aim of this study was to measure the magnitude of the difference wave between that of simultaneously presented stimuli and that of linearly added stimuli of separate responses. Participants were fitted with a dry-sensor EEG cap and were presented with a series of Stroop and auditory stimuli. For some Stroop stimuli, auditory stimuli occurred simultaneously or in a close time proximity to the Stroop stimuli. We sought to estimate the linear contribution of the ERP from Stroop and oddball stimuli. We found that the magnitude of the difference waves were 3.07 ± 1.65 μV and 2.82 ± 1.34 μV for congruent and incongruent stimuli, respectively. As the average amplitude in the P3 region for both the congruent and incongruent difference waves was lower than the magnitude of the auditory oddball presented simultaneously with Stroop stimuli (12.13 ± 1.00 μV for congruent and 11.78 ± 1.05 μV for incongruent Stroop), we expect that the contribution of P3 auditory oddball would not mask a potential Stroop effect even if the timing of the auditory oddball stimuli were experimentally manipulated, a direction that we hope to explore in future work. In conclusion, we determine this paradigm is suitable for measuring cognitive load in precisely timed tasks.Clinical Relevance- This study establishes the efficacy of presenting a Stroop task as a proxy for a cognitive challenge that could cause cognitive overload.


Ability Emotional Intelligence and Subjective Happiness in Adolescents: The Role of Positive and Negative Affect.
Adolescence is an increasingly vulnerable period for the onset of affective disorders and other mental health issues that can significantly impact an individual's subjective well-being. This study aims to examine the relationship between emotional intelligence (ability EI), measured with a performance-based instrument, and Subjective Happiness in adolescents. It also explores the mediating role of positive (PA) and negative affect (NA) in this association and the moderating role of gender. The sample consisted of 333 first-year secondary school students from five centers in Spain, with an average age of 12.11 years (SD = 0.64), ranging from 11-14 years. Path analysis revealed an indirect effect (through NA and PA jointly) of Total Ability EI on Subjective Happiness and a positive direct effect that was observed only in females. Furthermore, this association was explored through various branches of ability EI. The results of this study suggest that interventions aimed at improving emotional abilities in adolescents while modulating the intensity of their emotions could significantly impact their overall well-being.


Macrophages inhibit neovascularization in a murine model of age-related macular degeneration.
Age-related macular degeneration (AMD) is the leading cause of blindness in people over 50 y of age in at least three continents. Choroidal neovascularization (CNV) is the process by which abnormal blood vessels develop underneath the retina. CNV develops in 10% of patients with AMD but accounts for up to 90% of the blindness from AMD. Although the precise etiology of CNV in AMD remains unknown, the macrophage component of the inflammatory response, which has been shown to promote tumor growth and support atherosclerotic plaque formation, is thought to stimulate aberrant angiogenesis in blinding eye diseases. The current theory is that macrophage infiltration promotes the development of neovascularization in CNV. We examined the role of macrophages in a mouse model of CNV. IL-10(-/-) mice, which have increased inflammation in response to diverse stimuli, have significantly reduced CNV with increased macrophage infiltrates compared to wild type. Prevention of macrophage entry into the eye promoted neovascularization while direct injection of macrophages significantly inhibited CNV. Inhibition by macrophages was mediated by the TNF family death molecule Fas ligand (CD95-ligand). Immune vascular interactions can be highly complex. Normal macrophage function is critical in controlling pathologic neovascularization in the eye. IL-10 regulates macrophage activity in the eye and is an attractive therapeutic target in order to suppress or inhibit CNV in AMD that can otherwise lead to blindness.