Modifiable lifestyle factors associated with fragility hip fracture: a systematic review and meta-analysis.
Among the various hip fracture predictors explored to date, modifiable risk factors warrant special consideration, since they present promising targets for preventative measures. This systematic review and meta-analysis aims to assess various modifiable risk factors. We searched four online databases in September 2017. We included studies that reported on modifiable lifestyle risk factors for sustaining fragility hip fractures. The quality of the included studies was assessed using the Newcastle-Ottawa Scale (NOS). The inclusion criteria consisted of (1) adult patients with osteoporotic hip fracture, (2) original study, (3) availability of full text articles in English, and (4) report of a modifiable lifestyle risk factor. Thirty-five studies, containing 1,508,366 subjects in total, were included in this study. The modifiable risk factors that were significantly associated with an increased risk of hip fracture were the following: weight < 58 kg (128 lbs) (pooled OR 4.01, 95% CI 1.62-9.90), underweight body mass index (BMI) (< 18.5) (pooled OR 2.83, 95% CI 1.82-4.39), consumption of ≥ 3 cups of coffee daily (pooled OR 2.27, 95% CI 1.04-4.97), inactivity (pooled OR 2.14, 95% CI 1.21-3.77), weight loss (pooled OR 1.88, 95% CI 1.32-2.68), consumption of ≥ 27 g (approx. > 2 standard drinks) alcohol per day (pooled OR 1.54, 95% CI 1.12-2.13), and being a current smoker (pooled OR 1.50, 95% CI 1.22-1.85). Conversely, two factors were significantly associated with a decreased risk of hip fracture: obese BMI (> 30) (pooled OR 0.58, 95% CI 0.34-0.99) and habitual tea drinking (pooled OR 0.72, 95% CI 0.66-0.80). Modifiable factors may be utilized clinically to provide more effective lifestyle interventions for at risk populations. We found that low weight and underweight BMI carried the highest risk, followed by high coffee consumption, inactivity, weight loss, and high daily alcohol consumption.


Expression and function of a CP339,818-sensitive K⁺ current in a subpopulation of putative nociceptive neurons from adult mouse trigeminal ganglia.
Trigeminal ganglion (TG) neurons are functionally and morphologically heterogeneous, and the molecular basis of this heterogeneity is still not fully understood. Here we describe experiments showing that a subpopulation of neurons expresses a delayed-rectifying K(+) current (IDRK) with a characteristically high (nanomolar) sensitivity to the dihydroquinoline CP339,818 (CP). Although submicromolar CP has previously been shown to selectively block Kv1.3 and Kv1.4 channels, the CP-sensitive IDRK found in TG neurons could not be associated with either of these two K(+) channels. It could neither be associated with Kv2.1 channels homomeric or heteromerically associated with the Kv9.2, Kv9.3, or Kv6.4 subunits, whose block by CP, tested using two-electrode voltage-clamp recordings from Xenopus oocytes, resulted in the low micromolar range, nor to the Kv7 subfamily, given the lack of blocking efficacy of 3 μM XE991. Within the group of multiple-firing neurons considered in this study, the CP-sensitive IDRK was preferentially expressed in a subpopulation showing several nociceptive markers, such as small membrane capacitance, sensitivity to capsaicin, and slow afterhyperpolarization (AHP); in these neurons the CP-sensitive IDRK controls the membrane resting potential, the firing frequency, and the AHP duration. A biophysical study of the CP-sensitive IDRK indicated the presence of two kinetically distinct components: a fast deactivating component having a relatively depolarized steady-state inactivation (IDRKf) and a slow deactivating component with a more hyperpolarized V1/2 for steady-state inactivation (IDRKs).


Towards Faster Gene Expression Prediction via Dimensionality Reduction and Feature Selection.
The majority of genes have a genetic component to their expression. Elastic nets have been shown effective at predicting tissue-specific, individual-level gene expression from genotype data. We apply principal component analysis (PCA), linkage disequilibrium pruning, or the combination of the two to reduce, or generate, a lower-dimensional representation of the genetic variants used as inputs to the elastic net models for the prediction of gene expression. Our results show that, in general, elastic nets attain their best performance when all genetic variants are included as inputs; however, a relatively low number of principal components can effectively summarize the majority of genetic variation while reducing the overall computation time. Specifically, 100 principal components reduce the computational time of the models by over 80% with only an 8% loss in R2. Finally, linkage disequilibrium pruning does not effectively reduce the genetic variants for predicting gene expression. As predictive models are commonly made for over 27,000 genes for more than 50 tissues, PCA may provide an effective method for reducing the computational burden of gene expression analysis.


Complications After Hand-Assisted Laparoscopic Living Donor Nephrectomy.
To study the complications of hand-assisted laparoscopic living donor nephrectomy (HALLDN) with an emphasis on complications occurring early after hospital discharge up to 120 days after surgery. We retrospectively categorized complications using the Clavien-Dindo classification in 3002 HALLDNs performed at 1 center from January 1, 2000, through December 31, 2019. In addition to overall summaries, modeling was used to identify correlates of complications before and after living donation. Of these donors, 87% were White, 59% were female, the mean age was 45 years (range, 18-77 years), 30.3% had a body mass index of at least 30, and 36.3% had previous abdominopelvic surgery. There were no deaths related to the surgery. The incidence of major complications (intraoperative complications plus Clavien-Dindo grade ≥III postoperatively) was 2.5% (n=74). The overall complication rate was 12.4% (n=371), including 15 intraoperative, 76 postoperative before discharge, and 280 after discharge to 120 days. Reoperation was required in 1.8% of patients (n=54), and all but 1 of these were incision-related problems. Seventy-six percent of all complications occurred after discharge, including 85% of the reoperations. For major complications, no risk factor was found. Risk factors for any complication included paramedian incision (hazard ratio [HR], 2.54; 95% CI, 1.49 to 4.34; P<.001); a history of abdominopelvic surgery (HR, 1.37; 95% CI, 1.07 to 1.76; P=.01), male sex (HR, 1.37; 95% CI, 1.07 to 1.76; P=.01), non-White race (HR, 1.40; 95% CI, 1.05 to 1.88; P=.02), and early era of the experience. Most major complications of HALLDN occur after discharge, suggesting that close follow-up is warranted and that the current literature may underestimate the true incidence.


Early Virtual-Reality-Based Home Rehabilitation after Total Hip Arthroplasty: A Randomized Controlled Trial.
The benefits of early virtual-reality-based home rehabilitation following total hip arthroplasty (THA) have not yet been assessed. The aim of this randomized controlled study was to compare the efficacy of early rehabilitation via the Virtual Reality Rehabilitation System (VRRS) versus traditional rehabilitation in improving functional outcomes after THA. Subjects were randomized either to an experimental (VRRS; n = 21) or a control group (control; n = 22). All participants were invited to perform a daily home exercise program for rehabilitation after THA with different administration methods—namely, an illustrated booklet for the control group and a tablet with wearable sensors for the VRRS group. The primary outcome was the hip disability (HOOS JR). Secondary outcomes were the level of independence and the degree of global perceived effect of the rehabilitation program (GPE). Outcomes were measured before surgery (T0) and at the 4th (T1), 7th (T2), and 15th (T3) day after surgery. Mixed-model ANOVA showed no significant group effect but a significant effect of time for all variables (p < 0.001); no differences were observed in HOOS JR between VRRS and the control at T0, T1, T2, or T3. Further, no differences in the level of independence were found between VRRS and the control, whereas the GPE was higher at T3 in VRSS compared to the control (4.76 ± 0.43 vs. 3.96 ± 0.65; p < 0.001). Virtual-reality-based home rehabilitation resulted in similar improvements in functional outcomes with a better GPE compared to the traditional rehabilitation program following THA. The application of new technologies could offer novel possibilities for service delivery in rehabilitation.


PEG10 Promoter-Driven Expression of Reporter Genes Enables Molecular Imaging of Lethal Prostate Cancer.
The retrotransposon-derived paternally expressed gene 10 (PEG10) protein is ordinarily expressed at high levels in the placenta. Recently, it was discovered that PEG10 isoforms promote the progression of prostate cancer to a highly lethal androgen receptor (AR)-negative phenotype. The presence of PEG10 in other subtypes of prostate cancer has not been explored and a utility for PEG10 overexpression has not been developed. Here, we found that in addition to AR-null disease, PEG10 was also expressed in prostate cancer with constitutively active AR-splice variants. A molecular genetic imaging strategy for noninvasive imaging of AR-splice variant prostate cancer was developed by utilizing the cancer specificity of the PEG10 promoter to drive the expression of reporter genes. Plasmid insertion of a PEG10 promoter sequence optimized for enhanced output upstream of a reporter gene allowed detection of prostate cancer by near-infrared and positron emission tomography imaging after systemic administration of the plasmid in vivo. PEG10 expressing subcutaneous xenograft and intratibial tumor models were imaged by both modalities using this molecular genetic imaging strategy. This study demonstrates a preclinical proof-of-concept that the PEG10 promoter is a powerful and specific tool that can be utilized for noninvasive detection of aggressive prostate cancer subtypes. SIGNIFICANCE: PEG10 is expressed by prostate cancer with constitutively active AR-splice variants that can be exploited for noninvasive molecular imaging of this aggressive prostate cancer subytpe.


The reversible heat denaturation of chymotrypsinogen.
Within a restricted range of pH and protein concentration crystalline chymotrypsinogen undergoes thermal denaturation which is wholly reversed upon cooling. At a given temperature an equilibrium exists between native and reversibly denatured protein. Within the pH range 2 to 3 the amount of denatured protein is a function of the third power of the hydrogen ion activity. The presence of small amounts of electrolyte causes aggregation of the reversibly denatured protein. A specific anion effect has been observed at pH 2 but not at pH 3. Both the reversible denaturation reaction and the reversal reaction have been found to be first order reactions with respect to protein and the kinetic and thermodynamic constants for both reactions have been approximated at pH 2 and at pH 3. Renatured chymotrypsinogen has been found to be identical with native chymotrypsinogen with respect to crystallizability, solubility, activation to delta-chymotrypsin, sedimentation rate, and behavior upon being heated. Irreversible denaturation of chymotrypsinogen has been found to depend on pH, temperature, protein concentration, and time of heating. Irreversible denaturation results in an aggregation of the denatured protein.


Image design: generation of a prescribed image at the output of a band-limited system.
Image design involves determining the object distribution which produces a prescribed image at the output of a given imaging system. We solve the problem of generating two parallel lines at the output of an imaging system that is modeled by a linear band-limited system foliowed by a point nonlinearity of the hardlimiting type. It is shown that a solution always exists whatever the system's bandwidth. In principle, infinite two-line resolution is achievable. In practice, resolution much higher than the conventional resolution of the imaging system is possible.


Recovery of consciousness after severe brain injury: the role of arousal regulation mechanisms and some speculation on the heart-brain interface.
Recovery of consciousness after severe brain injury involves reconstitution of brain arousal mechanisms and cerebral integrative function. This review discusses several aspects of neuroanatomy and neuropathology relevant to the process of recovery. Particular emphasis is placed on the role of the anterior forebrain and circuit mechanisms linking the frontal lobe, striatum, and central thalamus. The article concludes with some observations on the heart-brain interface and future research directions in the context of recovery from severe brain injury.


Children's age at parental divorce and depression in early and mid-adulthood.
This study aimed to assess whether children's age at their parents' divorce is associated with depression in early and mid-adulthood, as indicated by medication purchase. A sibling comparison method was used to control for unobserved factors shared between siblings. The data were extracted from the Norwegian Population Register and Norwegian Prescription Database and included about 181,000 individuals aged 20-44 who had experienced parental divorce and 636,000 who had not. Controlling for age in 2004, sex, and birth order, children who were aged 15-19 when their parents divorced were 12 per cent less likely to purchase antidepressants as adults in 2004-08 than those experiencing the divorce aged 0-4. The corresponding reduction for those aged 20+ at the time of divorce was 19 per cent. However, the association between age at parental divorce and antidepressant purchases was only evident among women and those whose mothers had low education.


Autonomic Reactivity to Infant Crying in Maltreating Mothers.
We examined autonomic reactivity to infant crying in a sample of 42 maltreating and 38 non-maltreating mothers. Exploratively, we tested if differential reactivity was related to child neglect versus the combination of neglect and abuse, and we tested whether mothers' experiences with maltreatment in their own childhood moderated the association between their current maltreatment status and physiology. During a standardized cry paradigm, mothers listened to cry sounds of various pitches. Heart rate (HR), pre-ejection period (PEP), skin conductance levels (SCLs), and vagal tone (root mean square of successive differences [RMSSD]) were measured as indicators of underlying sympathetic and parasympathetic reactivity. The maltreating mothers showed lower SCL reactivity to the cry sounds than non-maltreating mothers. Furthermore, significant negative correlations between HR and PEP in the non-maltreating group differed from nonsignificant correlations in the maltreating group, which suggests a lack of sympathetic cardiac control in maltreating mothers. We found no differences between neglectful mothers and those who were additionally abusive. Together, our findings support the notion of sympathetic hypoarousal as a risk factor for child maltreatment, which may be indicative of disengagement in a caregiving context. Intervention programs might focus on improving maternal sensitivity to improve responsiveness to child signals.


Generalized pustular psoriasis in childhood.
Generalized pustular psoriasis is a rare form of psoriasis consisting of a generalized eruption of sudden onset with erythema and sterile pustules. In children, generalized pustular psoriasis is even more uncommon and may present as a severe and potentially life-threatening disorder. In this study, we present demographics, clinical aspects, treatment response, and follow-up of seven children with generalized pustular psoriasis. Retrospective study reviewing the records of seven children with generalized pustular psoriasis including age, gender, age of onset, presence of scalp and nail involvement, family history, concomitant diseases, precipitating factors, treatment modalities, and outcome. Age of first symptoms ranged from 1 month to 11 years. All patients received systemic retinoids at one time of the follow-up period. Other treatment modalities included immunosuppressive drugs, biologics, phototherapy, and sulfasalazine. Two patients presented with severe constitutional illness, secondary infection and septic shock, including one fatal outcome. All further cases have remained free of recurrences for a mean period of up to 3 years. In our study, generalized pustular psoriasis presented a wide clinical spectrum in children ranging from mild, asymptomatic outbreaks to more severe, life-threatening episodes. One fatality was observed. Children generally responded well to systemic retinoids. Further studies and long-term follow-up periods are needed to define potential trigger factors, efficacy and safety of different treatment modalities in children with generalized pustular psoriasis.


Magnetic Focus Lateral Flow Sensor for Detection of Cervical Cancer Biomarkers.
We report on a magnetic focus lateral flow biosensor (mLFS) for ultrasensitive detection of protein biomarkers in a practical format. With valosin-containing protein as a target protein, we show that the developed mLFS concept could detect as low as 25 fg/mL with magnetic focus to enhance target capture efficiency to deliver a 106-fold improvement in sensitivity compared to that of conventional lateral flow (LF) systems. The conceptualized strategy utilizes a simple magnet placed beneath the three-dimensional printed LF device to concentrate the targets at the signal zone without any additional instrumentation. In addition, protein mixtures extracted from the tissue of cervical cancer patients was also utilized to validate the sensor. To investigate the effect of magnetic focus on sensitivity, surface-enhanced Raman spectroscopy and dark-field imaging was utilized to characterize the distribution and movement of Fe3O4 core-Au shell nanoprobes in a model LF strip. Our experiments show that the magnetic focus results in an increased interaction time between the magnetic probe-labeled targets and the capture antibody, yielding a higher capture efficiency, allowing for ultrasensitive detection of the target not possible before with LF. The proposed mLFS can be utilized to detect a range of trace protein biomarkers for early diagnosis and can be combined with diverse pretreatment and signal amplification steps to query complex samples.


Low-dosage cytarabine therapy for herpes zoster with pneumonia.
Disseminated herpes zoster infection is frequently fatal. Cytarabine (cytosine arabinoside), a drug of variable efficacy, has offered some hope in the treatment of this condition. In this case of disseminated herpes zoster infection with pneumonia, there was remarkable recovery after low-dosage cytarabine infusion. The findings suggest that doses of the drug not exceeding 30 mg/sq m of body surface per 24 hours (30 mg/sq m/24 hr) may well be virucidal but not cytohematotoxic.


General practitioner perception of prostate-specific antigen testing has improved, but more awareness of prostate cancer risk in younger patients is still awaited.
In 2006, a county-wide survey of general practitioners (GPs) in the United Kingdom (UK) identified a reluctance to refer younger men with abnormal prostate specific antigen (PSA) levels. Younger men have the most to gain from early-detection of prostate cancer (PCa), which remains a national government priority in the UK and around the world. We sought to assess changes in perception of abnormal PSA-values amongst UK GPs over the past 10 years. A total of 500 self-administered paper questionnaires were distributed to individually named GPs. One hundred and forty two responded (28.4%), representing a patient population of ∼600,000. A series of visual analogue questions assessed referral thresholds and understanding of risk factors related to the development of PCa. GPs with a median of 23-years experience responded. Although mean PSA threshold for referral to urology did fall between 2006 and 2016 in both the 45-year (5.42 ng/mL vs. 4.61 ng/mL P = 0.0003) and 55-year (5.81 ng/mL vs. 5.30 ng/mL P = 0.0164) age groups, the median referral values were unchanged. Significantly, referral thresholds quoted for younger men (<65 years) were considerably higher than recommended UK maximum PSA-levels. Using case-based scenarios, practitioners appeared more likely to refer older men with abnormal PSA values, with GPs reporting an average 56.2% likelihood of referring an asymptomatic 55-year-old with elevated age-adjusted PSA of 4.6 ng/mL. A total of 95.1% recognised a family history of PCa to be a potential risk factor but other at-risk categories were not so clearly understood. Awareness of abnormal PSA values in UK primary care is improving, but continues to lag behind the evidence. Strategies to disseminate knowledge of maximum PSA-values to GPs should focus especially on those for younger patients.


Botulinum Toxin in Migraine Treatment.
Since botulinum toxin might have a therapeutic effect on pain, many studies investigating the efficiency of botulinum toxin in headache treatment have been done. The most satisfying results were achieved by botulinum toxin type A (BoNT/A) in the treatment of chronic migraine. In this paper, we reviewed the clinical effectiveness of BoNT/A in migraine and included our clinical experience. In our ongoing pilot study, where we have repeated BoNT/A injections every 12 weeks, The difference in the Migraine Disability Assessment (MIDAS) scores between the first and the second injections was 61.1%; and between the first and the 3rd injections was found to be 65.72%.


Failure in Medical Practice: Human Error, System Failure, or Case Severity?
The success rate in medical practice will probably never reach 100%. Success rates depend on many factors. Defining the success rate is both a technical and a philosophical issue. In opposition to the concept of success, medical failure should also be discussed. Its causality is multifactorial and extremely complex. Its actual rate and its real impact are unknown. In medical practice, failure depends not only on the human factor but also on the medical system and has at its center a very important variable-the patient. To combat errors, capturing, tracking, and analyzing them at an institutional level are important. Barriers such as the fear of consequences or a specific work climate or culture can affect this process. Although important data regarding medical errors and their consequences can be extracted by analyzing patient outcomes or using quality indicators, patient stories (clinical cases) seem to have the greatest impact on our subconscious as medical doctors and nurses and these may generate the corresponding and necessary reactions. Every clinical case has its own story. In this study, three different cases are presented to illustrate how human error, the limits of the system, and the particularities of the patient's condition (severity of the disease), alone or in combination, may lead to tragic outcomes There is a need to talk openly and in a balanced way about failure, regardless of its cause, to look at things as they are, without hiding the inconvenient truth. The common goal is not to find culprits but to find solutions and create a culture of safety.


[Specific expression of the foreign gene regulated by the rice rbcS promoter in transgenic rice].
To use different types of promoters in transgenic rice research, the 5'-upstream regulation region of rice Rubisco small subunit gene (rbcS) was cloned from a Chinese cultivar Wuyunjing 8, and its sequences were confirmed by comparison with the known genome sequences of both japonica and indica rice. The cloned rbcS promoter was fused to the 5'-upstream of GUS (beta-glucuronidase) coding region in a binary vector, and introduced into rice by Agrogacterium-mediated transformation. The integration of the rbcS-GUS fusion gene in transgenic rice was confirmed by PCR analysis. The results of both histochemical staining and quantitative analysis of GUS activity showed that the expression level of GUS fusion gene was significantly stronger in leaf blade and sheath than in other organs of transgenic rice plants, and the GUS activity was restricted to the mesophyll cells of leaf tissue, which showed that the rice rbcS promoter could control not only the tissue- but also the cell-specific expression of foreign genes in transgenic rice. The present results also demonstrated that light induction had a significant effect on the enhancement of transgene's expression when regulated by the rice rbcS promoter in transgenic rice. Our results showed that the rice rbcS promoter might be very useful for the expression of target genes in transgenic rice, with particularly high efficiency in leaf tissues.


Avoidance of bird repellents by mice (Mus musculus).
It is believed that mammalian chemosensory irritants are not aversive to birds and vice versa. Nevertheless, few avian repellents have been tested against mammals. For that reason, we evaluated the efficacy of 1.0% w/v methyl anthranilate, orthoaminoacetophenone, 2-amino-4',5'-methoxyacetophenone, 2-methoxyacetophenone, and veratryl amine as mouse repellents in 3-hr no-choice drinking tests. Relative to ingestion of plain water, all test substances significantly reduced (P < 0.05) intake. Orthoaminoaceto-phenone was the most effective repellent, with intake reduced to levels statistically indistinguishable from zero.


Necessity of q-expectation value in nonextensive statistical mechanics.
In nonextensive statistical mechanics, two kinds of definitions have been considered for expectation value of a physical quantity: one is the ordinary definition and the other is the normalized q-expectation value employing the escort distribution. Since both of them lead to the maximum-Tsallis-entropy distributions of a similar type, it is of crucial importance to determine which the correct physical one is. A point is that the definition of expectation value is indivisibly connected to the form of generalized relative entropy. Studying the properties of the relative entropies associated with these two definitions, it is shown how the use of the escort distribution is essential. In particular, the Shore-Johnson theorem for consistent minimum cross-entropy (i.e., relative-entropy) principle is found to select the formalism with the normalized q-expectation value and to exclude the possibility of using the ordinary expectation value from nonextensive statistical mechanics.


Novel C-3-(N-alkyl-aryl)-aminomethyl rifamycin SV derivatives exhibit activity against rifampicin-resistant Mycobacterium tuberculosis RpoBS522L strain and display a different binding mode at the RNAP β-subunit site compared to rifampicin.
Antimicrobial resistance is a main concern in tuberculosis treatment and is often associated with the emergence of Mycobacterium tuberculosis strains resistant to rifampicin (RIF), which is one of the cornerstones of tuberculosis chemotherapy. In this study, aminoalkyl-aromatic ring tails were appended to the C3 position of rifamycin core to assess the role of C3 substitutions to the anti-mycobacterial activity of the rifamycin antibiotics. The typical hydrazone unit of RIF was replaced by an amino-alkyl linkage to connect the aromatic ring tails with the rifamycin naphthoquinone core. Eight novel C3-(N-alkyl-aryl)-aminoalkyl analogues of rifamycin SV were synthesised and screened in vitro against wild-type HR37Rv and "hypervirulent" HN-878 strains, and a panel of rifampicin-resistant M. tuberculosis clinical isolates carrying mutations at the 522, 531 and 455 positions of the rpoB gene (RpoBS522L, RpoBS531L and RpoBH455D strains). The analogues exhibited anti-tubercular activity against H37Rv and HN-878 at submicromolar or nanomolar concentrations, and against clinical H37Rv isolates bearing the S522L mutations at low micromolar concentration. Benzylamine moiety-including analogue 8 was as active as rifampicin against HN-878 with a MIC90 value of 0.02 μM, whereas 14 and 15, which included tryptamine and para-methyl-sulfonylbenzylamine C3-substituents, respectively, showed higher anti-tubercular activity (MIC90 = 3 μM) compared to rifampicin against the S522L mutated H37Rv strain. Detailed in silico analysis of different RNAP molecular systems predicted a distinct, possibly novel, binding mode for the new rifamycin analogues. These were found to occupy a different space in the binding pockets of both wild type and mutated RNAP proteins compared to that of rifampicin. Moreover, the molecular modelling experiments investigated the ability of the novel analogues aromatic tails to establish key interactions at the RNAP binding site. These interesting findings might pave the way for generating rifamycin analogues that can overcome anti-microbial resistance in M. tuberculosis.


Detection of airborne influenza a virus in experimentally infected pigs with maternally derived antibodies.
This study assessed whether recently weaned piglets with maternally derived antibodies were able to generate infectious influenza aerosols. Three groups of piglets were assembled based on the vaccination status of the dam. Sows were either non-vaccinated (CTRL) or vaccinated with the same (VAC-HOM) strain or a different (VAC-HET) strain to the one used for challenge. Piglets acquired the maternally derived antibodies by directly suckling colostrum from their respective dams. At weaning, pigs were challenged with influenza virus by direct contact with an infected pig (seeder pig) and clinical signs evaluated. Air samples, collected using a liquid cyclonic air collector, and individual nasal swabs were collected daily for 10 days from each group and tested by matrix real-time reverse transcriptase polymerase chain reaction (RRT-PCR) assay. Virus isolation and titration were attempted for air samples on Madin-Darby canine kidney cells. All individual pigs from both VAC-HET and CTRL groups tested positive during the study but only one pig in the VAC-HOM group was positive by nasal swab RRT-PCR. Influenza virus could not be detected or isolated from air samples from the VAC-HOM group. Influenza A virus was isolated from 3.2% and 6.4% air samples from both the VAC-HET and CTRL groups, respectively. Positive RRT-PCR air samples were only detected in VAC-HET and CTRL groups on day 7 post-exposure. Overall, this study provides evidence that recently weaned pigs with maternally derived immunity without obvious clinical signs of influenza infection can generate influenza infectious aerosols which is relevant to the transmission and the ecology of influenza virus in pigs.


Multimodal navigation in the functional microsurgical resection of intrinsic brain tumors located in eloquent motor areas: role of tractography.
Nowadays the role of microsurgical management of intrinsic brain tumors is to maximize the volumetric resection of the tumoral tissue, minimizing the postoperative morbidity. The purpose of this paper was to study the benefits of an original protocol developed for the microsurgical treatment of tumors located in eloquent motor areas where the navigation and electrical stimulation of motor subcortical pathways have been implemented. A total of 17 patients who underwent resection of cortical or subcortical tumors in motor areas have been included in the series. The preoperative planning for multimodal navigation was done by integrating anatomical studies, motor functional MR (fMR) imaging, and subcortical pathway volumes generated by diffusion tensor (DT) imaging. Intraoperative neuromonitoring included motor mapping by direct cortical stimulation (CS) and subcortical stimulation (sCS), and localization of the central sulcus by using cortical multipolar electrodes and the N20 wave inversion technique. The location of all cortically and subcortically stimulated points with positive motor response was stored in the navigator and correlated with the cortical and subcortical motor functional structures defined preoperatively. The mean tumoral volumetric resection was 89.1 +/- 14.2% of the preoperative volume, with a total resection (> or = 100%) in 8 patients. Preoperatively a total of 58.8% of the patients had some kind of motor neurological deficit, increasing 24 hours after surgery to 70.6% and decreasing to 47.1% at 1 month later. There was a great correlation between anatomical and functional data, both cortically and subcortically. A total of 52 cortical points submitted to CS had positive motor response, with a positive correlation of 83.7%. Also, a total of 55 subcortical points had positive motor response; in these cases the mean distance from the stimulated point to the subcortical tract was 7.3 +/- 3.1 mm. The integration of anatomical and functional studies allows a safe functional resection of the brain tumors located in eloquent areas. Multimodal navigation allows integration and correlation among preoperative and intraoperative anatomical and functional data. Cortical motor functional areas are anatomically and functionally located preoperatively thanks to MR and fMR imaging and subcortical motor pathways with DT imaging and tractography. Intraoperative confirmation is done with CS and N20 inversion wave for cortical structures and with sCS for subcortical pathways. With this protocol the authors achieved a good volumetric resection in cortical and subcortical tumors located in eloquent motor areas, with an increase in the incidence of neurological deficits in the immediate postoperative period that significantly decreased 1 month later. Ongoing studies must define the safe limits for functional resection, taking into account the intraoperative brain shift. Finally, it must be demonstrated whether this protocol has any long-term benefit for patients by prolonging the disease-free interval, the time to recurrence, or the survival time.


The PRO-HOME Project. A multicomponent intervention for the protected discharge from the hospital of multimorbid and polytreated older individuals by using innovative technologies: A pilot study.
Discharge planning from the hospital of frail older patients is an important step to avoid inappropriate long-stay hospitalizations and to prevent the risks related to the prolonged hospitalization. In this frame, we developed an experimental trial-'PRO-HOME', a multicomponent programme of interventions for multimorbid and polytreated hospitalized older patients. The main aim of the study was to develop a protected discharge facility using a mini apartment equipped with advanced architectural and technological components to reduce the length of hospital stay of older participants (aged 65+ years old) admitted to the hospital for an acute event, deemed stable and dischargeable. This is a pilot randomized controlled study, comparing 30 hospitalized participants included in a multidimensional, transitional care programme based on information and communication technologies to 30 patients in standard usual care until hospital discharge. We presented the study design of the PRO-HOME programme, including architectural and technological components, the enrolment procedures, the components of the intervention that is physical activity, cognitive training and life-style education and the evaluation method of the intervention based on the Comprehensive Geriatric Assessment to explore the changes in the individual domains that are target of the multicomponent intervention. The final results will suggest whether the PRO-HOME programme represents a useful and feasible intervention to reduce the length of hospital stay of multimorbid and polytreated hospitalized older patients and improve their physical and cognitive performances and overall quality of life. Due to the characteristics of the population of interest of the PRO-HOME study, we involved in the study design and programme of the activities the participants enrolled in a previous smart home-based project named MoDiPro carried-out during a 3-year period. The elderly participants from the local population involved were asked, by means of focus groups, for feedback on their experience in MoDiPro, and their suggestions were integrated into the design phase of the current PRO-HOME project. The focus groups included open group interviews with a qualitative collection of the patients' feedback so that the participants could interact with each other.


Diagnostic and prognostic performance of the SAFE score in non-alcoholic fatty liver disease.
The steatosis-associated fibrosis estimator (SAFE) score was developed to detect clinically significant liver fibrosis in patients with NAFLD in the United States. We compare the performance of the SAFE score and other non-invasive tests to diagnose liver fibrosis and to correlate the scores with liver-related outcomes in patients with NAFLD in Hong Kong. This was a retrospective cohort study involving two data sets. The first cohort was a biopsy cohort of NAFLD patients (n = 279), and the second was a territory-wide cohort of NAFLD patients (n = 4603) retrieved from a territory-wide electronic healthcare database in Hong Kong. In detecting significant fibrosis, liver stiffness measured by transient elastography had the highest area under the receiver operating characteristic curve (AUROC) (.844), followed by SAFE score (.773). SAFE score had the highest AUROC among blood-based algorithms (.773 vs. .746 for FIB-4, .697 for APRI). Based on cut-off values of SAFE score (0 and 100 points), 854 (18.6%), 1596 (34.6%) and 2153 (46.8%) were in the low-, intermediate- and high-risk groups, respectively, in the territory-wide cohort. Six (.7%), 15 (.9%) and 59 (2.7%) developed liver-related events in those three groups respectively. Among patients who had liver-related events at 5 years, using the high cut-off, SAFE score could predict 84.9% of patients accurately, compared to 40.9% for FIB-4 and 27.2% for APRI. The SAFE score performed well and better than other blood-based markers in diagnosing significant fibrosis and predicting liver-related events in Asian patients with NAFLD.


Long-term enhancement of CA1 synaptic transmission is due to increased quantal size, not quantal content.
Quantal components of Schaffer collateral synaptic transmission recorded intracellularly from CA1 pyramidal cells were examined using 2 methods: simultaneous recordings of CA3-CA1 cell-pairs, and minimal electrical stimulation in stratum radiatum. Quantal parameters estimated by the method of failures and by a computer algorithm that optimized parameter estimates using deconvolution of background noise were highly correlated. EPSP-amplitude histograms of CA3-CA1 cell pairs (N = 10) and minimal electrical stimulation (N = 33) could be adequately described either by Poisson or binomial statistics, or by both, and exhibited similar estimates of unit quantal size (q) and mean quantal content (m). Paired-pulse stimulation with 50 msec between stimuli resulted in an expected facilitation in the EPSP amplitude and increase in m during the second response, as estimated by noise deconvolution, by the decrease in apparent failures, and by a decrease in the coefficient of variation of the EPSP. Tetanization of the Schaffer collaterals that induced long-term enhancement (LTE/LTP) of the population response was associated with an average increase in q for minimal-stimulation responses, with no significant change in any estimate of m. Taken together, these data indicate that, under the present experimental conditions, LTE is expressed as an increase in quantal size, rather than an increase in the number of quanta released per presynaptic impulse. Although this is not definitive evidence for a postsynaptic mechanism, these findings do further restrict the classes of possible presynaptic mechanisms that may be proposed to account for LTE expression.


Demonstration of ipsilateral retinocollicular projections in the tree shrew (Tupaia glis).
Ipsilateral retinocollicular projections labeled by anterograde transport of wheatgerm agglutinin-horseradish peroxidase (HRP) conjugate in the tree shrew were examined. For those animals in which this pathway was demonstrated (4 of 14) ipsilateral collicular labeling extended across approximately the anterior two-thirds of the colliculus, with the exception of the extreme rostral pole. Labeling was invariably punctuate and spaced at regular intervals in the lower stratum griseum superficiale. The laminar distribution and patchy terminations of ipsilateral projections are discussed in relation to two apparently independent pathways originating in the temporal retina, the crossed and uncrossed collicular pathways.


Maternal inheritance of human mitochondrial DNA.
Human mitochondrial DNA was obtained from peripheral blood platelets donated by the members of several independent families. The samples were screened for nucleotide sequence polymorphisms between individuals within these families. In each family in which we were able to detect a distinctly different restriction endonuclease cleavage pattern between the parents, the progeny exhibited the maternal cleavage pattern. Informative polymorphisms were detected for Hae II (PuGCGCPy) in a three-generation family composed of 33 members, for HincII (GTPyPuAC) in a two-generation family composed of four members, and for Hae III(GGCC) in a two-generation family composed of four members. The Hae II polymorphism was analyzed through all three generations in both the maternal and paternal lines. The results of this study demonstrate that human mitochondrial DNA is maternally inherited. The techniques described for using peripheral blood platelets as a source of human mitochondrial DNA represent a convenient way to obtain data on mitochondrial DNA variation in both individuals and populations.


Preventive Effect of Changing Contrast Media in Patients With A Prior Mild Immediate Hypersensitivity Reaction to Gadolinium-Based Contrast Agent.
Currently, the prevention of recurrent immediate hypersensitivity reactions (HSRs) to contrast media (CM) requests premedication and changing the culprit contrast agent. However, strategies for the prevention of immediate HSRs to gadolinium-based magnetic resonance contrast agents (GBCAs) have not yet been established. This study aimed to evaluate the effectiveness of changing the contrast agent and single-dose premedication for HSR recurrence prevention in patients with a history of mild immediate HSR to GBCA. The outcomes of patients with mild immediate HSR to GBCA who subsequently underwent enhanced magnetic resonance imaging between October 2012 and July 2017 were analyzed. The institutional CM monitoring system was retrospectively reviewed, and data on the application of premedication and choice of CM were obtained. Gadolinium-based magnetic resonance contrast agents were classified into 3 classes according to their molecular structure (macrocyclic ionic, macrocyclic nonionic, and linear ionic). Intravenous chlorpheniramine 4 mg, 30 minutes before the GBCA administration, or intravenous methylprednisolone sodium succinate 40 mg plus chlorpheniramine 4 mg, 1 hour before the GBCA administration, was administrated as premedication regimen. Recurrence rates of immediate HSR were compared according to prevention strategies. A total of 185 patients with a history of mild immediate HSR to GBCA were re-exposed to GBCA 397 times during the study period. The overall recurrence rate was 19.6% (78/397). Changing the culprit GBCA significantly reduced the recurrence rate, compared with reusing the culprit GBCA (6.9%, 9/130 and 25.8%, 69/267; P < 0.001). The recurrence rate was lowest when the GBCA was changed to a different molecular structure class from the culprit agent, followed by changing to CM with the same molecular structure and reusing the culprit GBCA (6.2%, 7/113 vs 11.8%, 2/17 vs 25.8%, 69/267; P < 0.001 with χ test for trend). Single-dose premedication demonstrated no significant prophylactic effect on recurrence (20.4%, 17/98 vs 17.3%, 61/299 with and without premedication, respectively; P = 0.509). Premedication in addition to changing CM also showed no additional prophylactic effect (7.2%, 7/97 and 6.1%, 2/33, respectively; P = 0.821). Changing the CM from the culprit agent could reduce the chance of HSR recurrence in patients with prior mild immediate HSR to GBCA, especially when the CM was changed to one of a different molecular structure class. However, single-dose premedication administration did not show significant HSR recurrence rate difference.


Mode of action of amphotericin B on chick embryo fibroblasts and on mouse Ehrlich tumour cells: a cytological and cytochemical analysis.
Chick embryo fibroblasts cultivated in vitro and Ehrlich ascites tumor cells (in vivo or in vitro) have been treated with amphotericin B. Cell multiplication is strongly inhibited. Large clear zones appear in the fibroblast nucleoi (phase-contrast and electron-microscope observations). Many treated fibroblasts and tumour cells have a high DNA content (pre-mitotic or polyploid level; measurements by cytophotometry). However, the RNA content (cytophotometry) and the total protein content (cytophotometry and micro-interferometry) are relatively low in the tumour cells. As shown by autoradiography, DNA synthesis is active but RNA synthesis and, in some cases, protein synthesis are inhibited. Due to this unbalanced growth, the cells cannot divide.


The effects of vertically coronally advanced flap and free gingival graft techniques on shallow vestibule: a randomized comparative prospective trial.
The present study aimed to compare the influence of vertically coronally advanced flap (V-CAF) and free gingival graft (FGG) techniques on shallow vestibule depth (VD). Parallel-arm randomized clinical trial was conducted on 38 sites treated with either FGG or V-CAF. Periodontal variables (VD, recession depth and width, probing depth, clinical attachment level, keratinized tissue height (KTH), and tissue thickness (TT)), clinician- and patient-based subjective variables were assessed. All periodontal variables showed significant improvements in both groups at all follow-up intervals compared to baseline (p < 0.05). Both groups increased VD compared to baseline. RC and CRC were similar after treatment for both techniques. FGG provided a greater increase in KTH (p < 0.001) and VD (VD1, p = 0.02 and VD2, p < 0.001) while V-CAF exhibited more TT gain (p = 0.002). Except overall tissue appearance that was better in V-CAF (p < 0.001), no inter-group significant difference existed in patient-based variables. Both techniques were significantly effective in VD increasing. While both techniques were equally successful in RC, V-CAF provided higher TT gain and better tissue appearance. V-CAF can be chosen instead of FGG in the treatment of recessions with shallow VD. It can be recommended to prefer V-CAF instead of FGG in the treatment of shallow vestibule. NCT05777811 (clinicaltrials.gov).


Perceptions of Ethical Decision-Making Climate Among Clinicians Working in European and U.S. ICUs: Differences Between Nurses and Physicians.
To examine perceptions of nurses and physicians in regard to ethical decision-making climate in the ICU and to test the hypothesis that the worse the ethical decision-making climate, the greater the discordance between nurses' and physicians' rating of ethical decision-making climate with physicians hypothesized to rate the climate better than the nurses. Prospective observational study. A total of 68 adult ICUs in 13 European countries and the United States. ICU physicians and nurses. None. Perceptions of ethical decision-making climate among clinicians were measured in April-May 2014, using a 35-items self-assessment questionnaire that evaluated seven factors (empowering leadership by physicians, interdisciplinary reflection, not avoiding end-of-life decisions, mutual respect within the interdisciplinary team, involvement of nurses in end-of-life care and decision-making, active decision-making by physicians, and ethical awareness). A total of 2,275 nurses and 717 physicians participated (response rate of 63%). Using cluster analysis, ICUs were categorized according to four ethical decision-making climates: good, average with nurses' involvement at end-of-life, average without nurses' involvement at end-of-life, and poor. Overall, physicians rated ethical decision-making climate more positively than nurses (p < 0.001 for all seven factors). Physicians had more positive perceptions of ethical decision-making climate than nurses in all 13 participating countries and in each individual participating ICU. Compared to ICUs with good or average ethical decision-making climates, ICUs with poor ethical decision-making climates had the greatest discordance between physicians and nurses. Although nurse/physician differences were found in all seven factors of ethical decision-making climate measurement, the factors with greatest discordance were regarding physician leadership, interdisciplinary reflection, and not avoiding end-of-life decisions. Physicians consistently perceived ICU ethical decision-making climate more positively than nurses. ICUs with poor ethical decision-making climates had the largest discrepancies.


Laser cooling in a silica optical fiber at atmospheric pressure.
For the first time, to the best of our knowledge, laser cooling is reported in a silica optical fiber. The fiber has a 21-µm diameter core doped with 2.06 wt.% ${{\rm Yb}^{3 + }}$Yb3+ and co-doped with ${{\rm Al}_2}{{\rm O}_3}$Al2O3 and ${{\rm F}^ - }$F- to increase the critical quenching concentration by a factor of 16 over the largest reported values for the Yb-doped silica. Using a custom slow-light fiber Bragg grating sensor, temperature changes up to $ - {50}\;{\rm mK}$-50mK were measured with 0.33 W/m of absorbed pump power per unit length at 1040 nm. The measured dependencies of the temperature change on the pump power and the pump wavelength are in excellent agreement with predictions from an existing model, and they reflect the fiber's groundbreaking quality for the radiation-balanced fiber lasers.


Non-linear Amplification of Variability Through Interaction Across Scales Supports Greater Accuracy in Manual Aiming: Evidence From a Multifractal Analysis With Comparisons to Linear Surrogates in the Fitts Task.
Movement coordination depends on directing our limbs to the right place and in the right time. Movement science can study this central requirement in the Fitts task that asks participants to touch each of two targets in alternation, as accurately and as fast as they can. The Fitts task is an experimental attempt to focus on how the movement system balances its attention to speed and to accuracy. This balance in the Fitts task exhibits a hierarchical organization according to which finer details (e.g., kinematics of single sweeps from one target to the other) change with relatively broader constraints of task parameters (e.g., distance between targets and width of targets). The present work seeks to test the hypothesis that this hierarchical organization of movement coordination reflects a multifractal tensegrity in which non-linear interactions across scale support stability. We collected movement series data during a easy variant of the Fitts task to apply just such a multifractal analysis with surrogate comparison to allow clearer test of non-linear interactions across scale. Furthermore, we test the role of visual feedback both in potential and in fact, i.e., by manipulating both whether experimenters instructed participants that they might potentially have to close their eyes during the task and whether participants actually closed their eyes halfway through the task. We predict that (1) non-linear interactions across scales in hand movement series will produce variability that will actually stabilize aiming in the Fitts task, reducing standard deviation of target contacts; (2) non-linear interactions across scales in head sway will stabilize aiming following the actual closing eyes; and (3) non-linear interactions across scales in head sway and in hand movements will interact to support stabilizing effects of expectation about closing eyes. In sum, this work attempts to make the case that the multifractal-tensegrity hypothesis supports more accurate aiming behavior in the Fitts task.


Hepatitis E and blood donation safety in selected European countries: a shift to screening?
The public health implications of hepatitis E virus (HEV) in Europe have changed due to increasing numbers of hepatitis E cases and recent reports of chronic, persistent HEV infections associated with progression to cirrhosis in immunosuppressed patients. The main infectious risk for such immunosuppressed patients is exposure to undercooked infected pork products and blood transfusion. We summarised the epidemiology of HEV infections among blood donors and also outlined any strategies to prevent transfusion-transmitted HEV, in 11 European countries. In response to the threat posed by HEV and related public and political concerns, most of the observed countries determined seroprevalence of HEV in donors and presence of HEV RNA in blood donations. France, Germany, Spain and the United Kingdom (UK) reported cases of transfusion-transmitted HEV. Ireland and the UK have already implemented HEV RNA screening of blood donations; the Netherlands will start in 2017. Germany and France perform screening for HEV RNA in several blood establishments or plasma donations intended for use in high-risk patients respectively and, with Switzerland, are considering implementing selective or universal screening nationwide. In Greece, Portugal, Italy and Spain, the blood authorities are evaluating the situation. Denmark decided not to implement the HEV screening of blood donations.


Prospective study of motor, sensory, psychologic, and autonomic functions in patients with irritable bowel syndrome.
The aim of this study was to assess pathophysiology in irritable bowel syndrome (IBS). A total of 122 IBS patients (3 male) and 41 healthy females underwent the following: questionnaires (symptoms, psychology), autonomic function, gut transit, gastric volumes, satiation, rectal compliance, and sensation (thresholds and pain ratings) testing. Proportions of patients with abnormal (<10th and >90th percentiles) motor or sensory functions according to bowel symptoms (constipation [C], diarrhea [D], mixed [M),) pain/bloat, and number of primary symptoms were estimated. IBS subgroups (C, D, M) were similar in age, gastric and small-bowel transit, satiation, gastric volumes, rectal compliance, sensory thresholds, and pain ratings. IBS was associated with body mass index, somatic symptoms, and anxiety and depression scores. Significant associations were observed with colonic transit (IBS-C [P = .078] and IBS-D [P < .05] at 24 h; IBS-D [P < .01] and IBS-M [P = .056] at 48 h): 32% of IBS patients had abnormal colonic transit: 20.5% at 24 hours and 11.5% at 48 hours. Overall, 20.5% of IBS patients had increased sensation to distensions: hypersensitivity (<10th percentile thresholds) in 7.6%, and hyperalgesia (pain sensation ratings to distension >90th percentile for ratings in health) in 13%. Conversely, 16.5% of IBS patients had reduced rectal sensation. Pain greater than 6 times per year and bloating were not associated significantly with motor, satiation, or sensory functions. Endorsing 1 to 2 or 3 to 4 primary IBS symptoms were associated with abnormal transit and sensation in IBS. In tertiary referral (predominantly female) patients with IBS, colonic transit (32%) is the most prevalent physiologic abnormality; 21% had increased and 17% had decreased rectal pain sensations. Comprehensive physiologic assessment may help optimize management in IBS.


Comprehensive care in chronic obstructive pulmonary disease.
Once a diagnosis of COPD is suspected, history, physical examination, pulmonary function tests, chest roentgenogram, sputum analysis, and so forth, are useful to assess the severity of obstructive airways diseases. A comprehensive program of care is then outlined (Table 2). General measures include avoidance of infection and inhalants, humidification, and proper rest and diet. Appropriate medications may include bronchodilators, antibiotics, corticosteroids, cromolyn sodium, digitalis, and diuretics. Inhalation therapy as aerosols, IPPB, and supplemental oxygen may be indicated. Physical therapy with postural drainage, exercise reconditioning, and occupational therapy deserve attention. The day-to-day care of the vast majority of patients with COPD is managed by primary care physicians. This systematic approach to pulmonary rehabilitation will yield definite rewards. Patients will feel and perform better. They will note an improved exercise tolerance, leading to increased activities of daily living. They will experience reduction in the frequency and duration of hospitalization as well as a decrease in anxiety and depression with an improved quality of life.


The effects of nanoscale pits on primary human osteoblast adhesion formation and cellular spreading.
Current understanding of the mechanisms involved in ossesoinegration following implantation of a biomaterial has led to an emphasis being placed on the modification of material topography to control interface reactions. Recent studies have inferred nanoscale topography as an important mediator of cell adhesion and differentiation. Biomimetic strategies in orthopaedic research aim to exploit these influences to regulate cellular adhesion and subsequent bony tissue formation. Here experimental topographies of nanoscale pits demonstrating varying order have been fabricated by electron-beam lithography in (poly)carbonate. Osteoblast adhesion to these nanotopographies was ascertained by quantification of the relation between adhesion complex formation and total cell area. This study is specifically concerned with the effects these nanotopographies have on adhesion formation in S-phase osteoblasts as identified by BrdU incorporation. Nanopits were found to reduce cellular spreading and adhesion formation.


Vasoactive intestinal peptide modulates T lymphocyte migration in Peyer's patches of rat small intestine.
Although vasoactive intestinal peptide (VIP) has been postulated to function in modulation of T cell trafficking, the exact mechanism has not been elucidated in vivo. In the present study, the effects of VIP on T lymphocyte migration were examined in rat Peyer's patches. T lymphocytes collected from intestinal lymph of rats were labeled with carboxyfluorescein diacetate succinimidyl ester and injected into the jugular vein. Peyer's patches of the recipient rats were observed with intravital fluorescence microscopy. In vivo intra-arterial infusion of or in vitro incubation with VIP did not affect the initial lymphocyte interaction with postcapillary venules of Peyer's patches. However, these treatments with VIP significantly inhibited transendothelial migration and also significantly blocked the interstitial migration of T cells and inhibited their subsequent appearance in the interfollicular lymphatics. Treatment with adenosine 3',5'-cyclic monophosphate (cAMP)-inducing agents resulted in similar inhibitory effect on T lymphocyte migration in Peyer's patches. In conclusion, VIP has significant inhibitory effects on T lymphocyte migration in Peyer's patches, possibly mediated by elevation of the intracellular cAMP concentrations.


Neuroactive steroids and human recombinant rho1 GABAC receptors.
The gamma-aminobutyric acid type C (GABAC) receptor is structurally related to the GABAA receptors, yet quite distinct physiologically and pharmacologically. Neuroactive steroids are known to be potent and efficacious modulators of the GABAA receptor, but they are less well characterized in their actions on the GABAC receptor. We have examined the actions of neuroactive steroids and analogs on rho1 (GABAC) receptors expressed in Xenopus laevis oocytes, with two goals in mind. First, we tested a larger number of endogenous steroids, to determine whether particularly potent steroids could be found. Second, we examined the structure-activity relationship for steroid actions, and some mechanistic features, to determine the possible numbers of steroid binding sites and mechanisms of action. In total, 41 compounds were examined. Estradiols are inhibitors, essentially equipotent with picrotoxinin. No endogenous steroid tested was highly efficacious at potentiation. The results of the structure-activity studies and the effects of two mutations to the second transmembrane region of the rho1 GABAC receptor indicate that there are several mechanisms by which steroids can inhibit the receptor. Surprisingly, estradiol shares some features with picrotoxin. Inhibition by negatively charged compounds was not sensitive to membrane potential, and inhibition by all compounds tested was reduced at higher concentrations of GABA. The data indicate that the binding sites mediating potentiation and inhibition differ from each other and that there are several (three or more) mechanisms for producing inhibition.


Optimized High-Yield Purification of Obesity-Associated Melanocortin 4 Receptor.
Obesity has emerged as a global public health challenge associated with increased risk of hyperlipidemia and hypertension. It contributes to high sympathetic activity and increased catecholamine levels. The hypothalamic melanocortin system is known to regulate the energy homeostasis. The role of melanocortin 4 receptor (MC4R) has been demonstrated pharmacologically and in animal studies, which showed that severe obesity in MC4R knockout mice was caused by increased food intake and decreased energy consumption. Over 70 multiple different mis- -sense and nonsense mutations in hMC4R have been found at a high frequency of 2-8% in severe early onset or hereditary obesity. The single amino acid variation (D90N) located in the second transmembrane domain (TM2) of MC4R results in accelerated growth and childhood onset obesity. Interestingly, the functional characterization of D90N hMC4R mutant TM2 (m-hMC4R-TM2) revealed normal cell surface expression and binding with agonist similar to the hMC4R wild-type TM2 (wt-hMC4R-TM2) but loss of signal transduction mediated via Gs/adenylyl cyclase activation. It is essential to delineate the three-dimensional structure of MC4Rs in order to elucidate their functional aspects. In this study, we demonstrate the optimized expression and isolation of wt/m-hMC4R-TM2 proteins under different chemical cleavage reaction times and purification procedures via SDS precipitation. The solid-state NMR spectroscopy was carried out to study the structure of wt/m-hMC4R- TM2 protein in the anisotropic phospholipid bicelles. The KSI-wt/m-hMC4R-TM2 fusion proteins developed in cell culture with LB medium. In order to isolate the expressed fusion protein from the cell, ultrasonication, Ni-NTA affinity chromatography, dialysis, and lyophilization techniques were used. Then, to obtain a protein with higher purity and higher yield, the CNBr chemical cleavage time was subdivided into 30 minutes, 1 h, 2 h, 3 h, and 4 h. Purification process was performed using FPLC, and 100 mM KCl and dialysis were used to remove the SDS. CD spectrometer, MALDI-TOF, solution-state NMR, and solid-state NMR were used to confirmed purity and structure of the wt/m-hMC4R-TM2. The precipitation method was used to remove the SDS bound to proteins as KCl-SDS. We optimized the 2 h cleavage reaction times for both wt-hMC4R-TM2 and m-hMC4R-TM2 depending on the purity based on mass spectra and 1H-15N HSQC spectra and the yield after final purification. The 1D 1H-15N CP (Cross polarization) solid-state NMR spectra suggest that the wt/m-hMC4R- TM2 undergo rotational diffusion around a perpendicular axis along the bilayer normal. We expressed wt/m-hMC4R-TM2 in E.coli and optimized the isolation and purification process, especially CNBr chemical cleavage time. The efficiency of KCl-SDS precipitation was confirmed via MALDI-TOF MS and the pure proteins obtained using this method were characterized by CD spectroscopy and solution-state NMR. The results of 1H-15N HSQC spectra in solution- state NMR also show the probability for structural studies. The 1D 1H-15N CP solid-state NMR spectra indicate that most of the residues in both the wt/m-hMC4R-TM2 peptides are integrated into the membrane.


Long-term exposure to a butter-rich diet induces mild-to-moderate steatosis in Chang liver cells and Swiss albino mice models.
Butter is one of the widely used fats present in the diet. However, there is no satisfactory study available that evaluates the effect of a high-fat diet containing butter as the principal fat on the development of non-alcoholic fatty liver disease (NAFLD). In the present study, butter was used for the development of steatosis in Chang liver cells in an in vitro study and Swiss albino mice in an in vivo study. In vitro steatosis was established, and butter was compared with oleic acid in Chang liver cells using an oil red O (ORO)-based colorimetric assay. In the in vivo study, a butter-rich special diet was fed for 15 weeks to mice, who showed no significant change in body weight. The expression pattern of phosphatase and tensin homolog (PTEN) and miR-21 was compared by reverse transcriptase-PCR. Special diet-fed animals showed downregulated PTEN compared to normal diet-fed animals, while levels of miR-21 remained the same. Elevations in biochemical parameters, viz., triglycerides and liver function tests showed symptoms of onset of NAFLD. Histophathological study of livers of test animals confirmed mild-to-moderate degree of NAFLD.


Distinct structural features of phospholipids differentially determine ethanol sensitivity and basal function of BK channels.
Large conductance Ca2+ -activated K+ (BK) channel activity and its potentiation by ethanol are both critically modulated by bilayer phosphatidylserine (PS), a phospholipid involved in membrane-bound signaling. Whether PS is uniquely required for ethanol to modify channel activity is unknown. Furthermore, the structural determinants in membrane phospholipid molecules that control alcohol action remain to be elucidated. We addressed these questions by reconstituting BK channels from human brain (hslo) into bilayers that contained phospholipids differing in headgroup size, charge, and acyl chain saturation. Data demonstrate that ethanol potentiation of hslo channels is blunted by conical phospholipids but favored by cylindrical phospholipids, independently of phospholipid charge. As found with ethanol action, basal channel activity is higher in bilayers containing cylindrical phospholipids. Basal activity and its ethanol potentiation in bilayers containing phosphatidylcholine, however, are not as robust as in those containing PS. These results are best interpreted as resulting from the relief of bilayer stress caused by inclusion of cylindrical phospholipids, with this relief being synergistically evoked by molecular shape and negative headgroup charge. Present findings suggest that hslo gating structures targeted by ethanol are accessible to sense changes in bilayer stress. In contrast, hslo unitary conductance is significantly higher in bilayers that contain negatively charged phospholipids independently of molecular shape, a result that is likely to be dependent on an interaction between anionic phospholipids and deep channel residues coupled to the selectivity filter.


CXCL1: A new diagnostic biomarker for human tuberculosis discovered using Diversity Outbred mice.
More humans have died of tuberculosis (TB) than any other infectious disease and millions still die each year. Experts advocate for blood-based, serum protein biomarkers to help diagnose TB, which afflicts millions of people in high-burden countries. However, the protein biomarker pipeline is small. Here, we used the Diversity Outbred (DO) mouse population to address this gap, identifying five protein biomarker candidates. One protein biomarker, serum CXCL1, met the World Health Organization's Targeted Product Profile for a triage test to diagnose active TB from latent M.tb infection (LTBI), non-TB lung disease, and normal sera in HIV-negative, adults from South Africa and Vietnam. To find the biomarker candidates, we quantified seven immune cytokines and four inflammatory proteins corresponding to highly expressed genes unique to progressor DO mice. Next, we applied statistical and machine learning methods to the data, i.e., 11 proteins in lungs from 453 infected and 29 non-infected mice. After searching all combinations of five algorithms and 239 protein subsets, validating, and testing the findings on independent data, two combinations accurately diagnosed progressor DO mice: Logistic Regression using MMP8; and Gradient Tree Boosting using a panel of 4: CXCL1, CXCL2, TNF, IL-10. Of those five protein biomarker candidates, two (MMP8 and CXCL1) were crucial for classifying DO mice; were above the limit of detection in most human serum samples; and had not been widely assessed for diagnostic performance in humans before. In patient sera, CXCL1 exceeded the triage diagnostic test criteria (>90% sensitivity; >70% specificity), while MMP8 did not. Using Area Under the Curve analyses, CXCL1 averaged 94.5% sensitivity and 88.8% specificity for active pulmonary TB (ATB) vs LTBI; 90.9% sensitivity and 71.4% specificity for ATB vs non-TB; and 100.0% sensitivity and 98.4% specificity for ATB vs normal sera. Our findings overall show that the DO mouse population can discover diagnostic-quality, serum protein biomarkers of human TB.


FoxP2 regulation during undirected singing in adult songbirds.
Learned vocal communication, including human speech, is a socially influenced behavior limited to certain animals. This ability requires auditory feedback during vocalization, which allows for on-line evaluation, to achieve the desired vocal output. To date, FOXP2 (forkhead box P2), a transcriptional repressor, is the only molecule directly linked to human speech. Identified FOXP2 mutations cause orofacial dyspraxia accompanied by abnormalities in corticostriatal circuitry controlling voluntary orofacial movements. These observations implicate FOXP2 in the developmental formation of neural circuits used in speech, but whether FOXP2 additionally plays an active role in mature circuitry was unknown. To address this question, we use a songbird, the zebra finch (Taeniopygia guttata), whose learned song and underlying circuitry are well characterized. We show that, when adult males sing, FoxP2 mRNA is acutely downregulated within area X, the specific region of the songbird striatum dedicated to song. Furthermore, we find downregulation in males that sing by themselves (undirected singers) but not in males that sing to females (directed singers). This FoxP2 downregulation cannot be a simple consequence of the motor act because birds sang in both directed and undirected contexts. Our data suggest that FoxP2 is important not only for the formation but also for the function of vocal control circuitry. Social context-dependent, acute changes in FoxP2 within the basal ganglia of adult songbirds also suggest, by analogy, that the core deficits of affected humans extend beyond development and beyond basic central motor control.


[Cytological mechanisms involved in assimilation of n-alkanes by yeasts].
The paper describes cytological mechanism of adaptation of yeasts to assimilation of aliphatic hydrocarbons added to a growth medium as a sole source of carbon. The process was studied by light optical and electron microscopy, employing fluorescent labelling and electron microscopy contrasting. Two types of yeasts were found, which differed by the response of the cell walls to hydrocarbons: those that formed "channels" and those that did not form them. Cytological response to hydrocarbon assimilation was detected also in the mitochondria and canals of the endoplasmic reticulum. Components of the Golgi apparatus may also participate in this process, in particular, in formation of peroxisomes (microbodies). Close contacts of the yeast cells with the hydrocarbon being assimilated is important; assimilation may start in a close vicinity of the cell walls. The rate of flavin production by Candida tropicalis 303 IBFM increases during growth on solid paraffins, beginning with C20-paraffin.


Interferometric optical time-domain reflectometry for distributed optical-fiber sensing.
The technique of optical time-domain reflectometry is analyzed to determine the effect of an optical phase modulation on light backscattered in an optical fiber. It is shown that the spatial distribution along the fiber of an external phase modulation can be measured with a spatial resolution close to that of optical time-domain reflectometry. A distributed interferometric sensor arrangement that employs this technique is investigated experimentally, and a satisfactory interrogation of more than 1000 resolution intervals is demonstrated.


Intraexaminer reliability of identifying a dysfunctional segment in the thoracic and lumbar spine.
To examine the intrarater reliability of identifying a manipulable lesion in the lumbar and thoracic spine. An experienced osteopath used dynamic and static examination to assess 12 asymptomatic subjects for signs of joint dysfunction in the thoracic and lumbar spine. The selected segment was marked with an UV invisible mark. A second examiner visualized these marks with an UV lamp and recorded them on acetates for analysis; this process was then repeated an hour later. The distance from the marks to a fixed point was measured and within-day intrarater reliability was calculated using intraclass correlation coefficients (ICCs). The ICC(1,1) for the thoracic spine was 0.70 (95% confidence interval [CI], 0.27-0.90). In the lumbar spine the ICC(1,1) was 0.96 (95% CI, 0.87-0.99). This study shows that the lumbar spine joint perceived to be the joint most likely to benefit from a high-velocity low-amplitude thrust can be identified with good within-day reliability in an asymptomatic sample using a defined examination protocol. However, the reliability in identifying a joint exhibiting signs of segmental dysfunction in the thoracic spine was poor.


UXO victims and health status of populations living in Xiengkhuang Province, Lao PDR: A household-based survey.
During the Vietnam War from 1964 to 1973, over 2 million tons of bombs were dropped on Laos. Approximately 30% of the bombs did not explode and have posed a continued threat to civilians throughout the country. Approximately 200 casualties per year have been reported nationwide. Therefore, we conducted a household survey to better understand magnitude of UXO victims, accessibility to the MCH services and child healthcare seeking behaviors of the population in the Xiengkhuang province. The household-based survey was carried out in February 2006 among 6 of 541 villages. 283 household representatives were interviewed. The cumulative number of UXO victims identified from 1973 to 2005 was 45 casualties, of which 9 (20.0%) occurred in the year 2005. 37.5% reported knowledge of danger zones for UXO contamination. Among the 91 children under the age of 5 years, households on average reported 1.7 episodes per year for diarrhea, 2.7 for respiratory disease, 2.3 for fever and 1.7 for measles. 69.4% of children under five were completed the routine immunization program, 62.6% of mothers used ANC service for their most recent pregnancy, 58.2% accessed family planning services, and 28.6% delivered their most recent child at a healthcare facility. UXO victims in the targeted villages sharply increased in 2005. Insufficient knowledge about UXO danger and MRE seems to be a central factor in the high rate of UXO-associated accidents. Diarrhea, respiratory disease and measles remained health problems for children under 5 years. MCH services utilization were higher than Laos nationally.


Circulating miR-22, miR-24 and miR-34a as novel predictive biomarkers to pemetrexed-based chemotherapy in advanced non-small cell lung cancer.
Pemetrexed has been widely used in patients with advanced non-small cell lung cancer (NSCLC). The clinical relevance of polymorphisms of folate pathway genes for pemetrexed metabolism have not been fully elucidated yet. The aim of this study was to evaluate the expression levels of circulating miR-22, miR-24, and miR-34a, possibly involved in folate pathway, in NSCLC patients treated with pemetrexed compared with healthy controls and to investigate their impact on patient clinical outcomes. A total of 22 consecutive patients with advanced NSCLC, treated with pemetrexed-based chemotherapy and 27 age and sex matched healthy controls were included in this preliminary analysis. miR-22, miR-24, and miR-34a targets were identified by TargetScan 6.2 algorithm, validating the involvement of these microRNAs in folate pathway. MicroRNAs were isolated from whole blood and extracted with miRNAeasy Mini Kit (Qiagen). miRNA profiling was performed using Real-Time PCR. SPSS 17 was used to data analysis. miR-22, miR-24, and miR-34a were found upregulated (P<0.05) in NSCLC patients versus healthy controls. Higher expression levels were recorded for miR-34a. Nevertheless, significantly higher miR-22 expression was observed in patients developing progressive disease (P=0.03). No significant associations with clinical outcome were recorded for miR-24 and miR-34a. Albeit preliminary, these data support the involvement of miR-22, miR-24, and miR-34a in advanced NSCLC. The correlation between high expression of miR-22 in whole blood and the lack of response in pemetrexed treated NSCLC patients indicates that miR-22 could represent a novel predictive biomarker for pemetrexed-based treatment.


FACTORS ASSOCIATED WITH INDISCRIMINATE FRIENDLINESS IN HIGH-RISK CHILDREN.
Indiscriminate friendliness (IF) refers to a lack of reticence with strangers and is well-documented in neglected children. This risky behavior is distinct from attachment insecurity, and persists when parenting/caregiving improves. A previous review has suggested that caregiving quality is not associated with IF. This review aimed to explore factors associated with IF and whether quality of caregiving is important. Ten articles were reviewed using the S.H. Downs and N. Black (1998) Checklist for randomized and nonrandomized studies. Overall quality was high. Results showed that IF is present in fostered populations and postinstitutionalized children. Attachment security was not associated with IF. Length of time in institution and inhibitory control were associated with IF. Inhibitory control moderated the association between IF and number of caregivers. Genetic factors predispose children to IF and may impact on persistence. Quality of caregiving was associated with IF. Emotional availability (i.e., the degree to which carer and child are able to respond to each other's emotional signals) predicted IF. Limitations across studies included heterogeneity in IF measurement and unreliable measures of preadoptive care. Parenting may be a useful target for intervention. Future research should focus on developing a standardized measure of IF as well as evaluating a parental intervention.


Magnesium supplementation helps to improve carotid intima media thickness in patients on hemodialysis.
The atherosclerotic process progresses more dynamically in hemodialysis (HD) patients than in the general population. In HD patients, lower magnesium levels were reported to be associated with increased atherosclerosis of the common carotid artery. We tested the hypotheses that magnesium supplementation helps to improve carotid intima media thickness (IMT) in HD patients. A total of 47 patients on HD were included in the study. Patients were randomly divided into two groups: group A (Mg group), in which patients were given magnesium citrate orally at a dosage of 610 mg every other day for 2 months and group B (control group), in which patients received only calcium acetate therapy as a phosphate binder. At baseline and 2 months later, all patients underwent a carotid artery ultrasound scan to measure carotid IMT. At the end of 2 months, mean serum calcium, phosphorus, and calcium x phosphorus product were not changed in both groups. As expected, mean serum Mg level significantly increased in the Mg group at the end of 2 months. In addition, serum parathyroid hormone (PTH) level significantly decreased in the Mg group at the end of 2 months (P = 0.003). Baseline carotid IMT was similar between the groups. Bilateral carotid IMT was significantly improved in patients treated with magnesium citrate compared to initial values (P = 0.001 for left, P = 0.002 for right). Based on the present data, magnesium may play an important protective role in the progression of atherosclerosis in patients on dialysis. Further studies are needed to assess more accurately the role of magnesium in atherosclerotic regression in dialysis patients.


Optical logarithmic filtering using inherent film nonlinearity.
Linear optical spatial filtering cannot be effectively applied to multiplied and convolved signals. One approach is to first perform a logarithmic transformation to produce a signal in additive form suitable for linear filtering processes [A. V. Oppenheim et al., Proc. IEEE 56, 1264 (1968)]. It was first suggested that a halftone screen be used to perform such a transformation [H. Kato and J. W. Goodman, Appl. Opt. 14, 1813 (1975)]. However, the maximum spatial resolution of this halftone screen technique is limited by the resolution of the screen. In this paper, we shall propose a different technique using the inherent film nonlinearity for the logarithmic transformation. Such a technique would enable the transformation of signals of very high spatial resolution, limited only by the resolution limit of the photographic film. This technique is applied to the spatial filtering and detection of signals in multiplicative noise. Experimental comparisons between linear and logarithmic filtering are presented.


The Impact of Diabetes on Morbidity and Mortality Following Thyroidectomy.
To explore the association between diabetes and outcomes in thyroidectomy patients. This retrospective cohort analysis used the 2015-2017 American College of Surgeons National Surgery Quality Improvement Program database. Current Procedural Terminology (CPT) codes were used to identify thyroidectomy cases (60210, 60212, 60220, 60225, 60240, 60252, 60254, 60260, 60270, and 60271). Demographics, comorbidities, and complication incidences were compared between diabetic and nondiabetic patients using Pearson's chi-square test/Fisher's exact test as appropriate. The independent effect of diabetes on outcomes was analyzed using binary logistic regression. A total of 47,776 (95.4%) nondiabetic and 2307 (4.6%) diabetic patients undergoing thyroidectomy were identified from 2015 to 2017. Chi-square analysis demonstrated that diabetic patients had higher incidences of obesity (55.2% vs. 33.2%; p < 0.001), dyspnea (12.7% vs. 4.8%; p < 0.001), poor functional status (1.9% vs. 0.4%; p < 0.001), ventilator dependence (0.6% vs. 0.1%; p < 0.001), chronic obstructive pulmonary disease (COPD; 6.8% vs. 2.2%; p < 0.001), congestive heart failure (1.1% vs. 0.3%; p < 0.001), acute renal failure (0.3% vs. 0.0%; p < 0.001), hypertension (79.2% vs. 32.4%; p < 0.001), dialysis (2.0% vs. 0.4%; p < 0.001), open wound (1.1% vs. 0.1%; p < 0.001), steroid use (5.3% vs. 2.3%; p < 0.001), bleeding disorders (3.6% vs. 0.9%; p < 0.001), preoperative blood transfusions (0.2% vs. 0.0%; p = 0.001), and systemic sepsis (1.0% vs. 0.3%; p < 0.001). Demographic characteristics were significantly different between the cohorts including gender (p < 0.001), age (p < 0.001), race (p < 0.001), and Hispanic ethnicity (p = 0.033). After adjusting for these factors, logistic regression analyses showed that diabetes was associated with acute renal failure (OR: 5.836; 95% CI: 1.060-32.134; p = 0.043), wound disruption (OR: 6.194; 95% CI: 1.752-21.900; p = 0.005), prolonged length of stay (OR: 1.430; 95% CI: 1.261-1.622; p < 0.001), unplanned readmission (OR: 1.380; 95% CI: 1.096-1.737; p = 0.006), superficial incisional surgical site infections (OR: 0.240; 95% CI: 0.058-0.995; p = 0.049), urinary tract infection occurrences (OR: 2.173; 95% CI: 1.186-3.980; p = 0.012), organ space surgical site infection occurrences (OR: 3.322; 95% CI: 1.016-10.864; p = 0.047), pneumonia occurrences (OR: 2.091; 95% CI: 1.125-3.884; p = 0.020), any medical complication (OR: 1.697; 95% CI: 1.246-2.313; p = 0.001), and any complication (OR: 1.495; 95% CI: 1.136-1.968; p = 0.004). Diabetes mellitus is a significant factor associated with increased odds of complications following thyroidectomy. 3 Laryngoscope, 133:3628-3632, 2023.


Selective disruption of dopamine D2 receptors in pituitary lactotropes increases body weight and adiposity in female mice.
Prolactin, a pleiotropic hormone secreted by lactotropes, has reproductive and metabolic functions. Chronically elevated prolactin levels increase food intake, but in some hyperprolactinemic states such as in the global dopamine D2 receptor (D2R) knockout mouse, food intake is not increased. Here, we conduct a cell-specific genetic dissection study using conditional mutant mice that selectively lack D2Rs from pituitary lactotropes (lacDrd2KO) to evaluate the role of elevated prolactin levels without any confounding effect of central D2Rs on motor and reward mechanisms related to food intake. LacDrd2KO female mice exhibited chronic hyperprolactinemia, pituitary hyperplasia, and a preserved GH axis. In addition, lacDrd2KO female but not male mice showed increased food intake by 3 months of age, and from 5 months onward their body weights were heavier. Marked increments in fat depots, adipocyte size, serum triglycerides, and nonesterified fatty acid levels and a decrease in lipolytic enzymes in adipose tissue were seen. Furthermore, lacDrd2KO female mice had glucose intolerance but a preserved response to insulin. In the hypothalamus, Npy mRNA expression was increased, and Pomc and Ppo mRNA levels were unaltered (in contrast to results in global D2R knockout mice). Thus, the orexigenic effect of prolactin and its action on hypothalamic Npy expression were fully evidenced, leading to increased food intake and adiposity. Our results highlight the metabolic role of prolactin and illustrate the value of studying cell-specific mutant mice to disentangle the pathophysiological mechanisms otherwise masked in null allele mutants or in animals treated with pervasive pharmacological agents.


Chronic stress induces a selective decrease in AMPA receptor-mediated synaptic excitation at hippocampal temporoammonic-CA1 synapses.
Chronic stress promotes depression, but how it disrupts cognition and mood remains unknown. Chronic stress causes atrophy of pyramidal cell dendrites in the hippocampus and cortex in human and animal models, and a depressive-like behavioral state. We now test the hypothesis that excitatory temporoammonic (TA) synapses in the distal dendrites of CA1 pyramidal cells in rats are altered by chronic unpredictable stress (CUS) and restored by chronic antidepressant treatment, in conjunction with the behavioral consequences of CUS. We observed a decrease in AMPAR-mediated excitation at TA-CA1 synapses, but not Schaffer collateral-CA1 synapses, after CUS, with a corresponding layer-specific decrease in GluA1 expression. Both changes were reversed by chronic fluoxetine. CUS also disrupted long-term memory consolidation in the Morris water maze, a function of TA-CA1 synapses. The decreases in TA-CA1 AMPAR-mediated excitation and performance in the consolidation test were correlated positively with decreases in sucrose preference, a measure of anhedonia. We conclude that chronic stress selectively decreases AMPAR number and function at specific synapses and suggest that this underlies various depressive endophenotypes. Our findings provide evidence that glutamatergic dysfunction is an underlying cause of depression and that current first-line antidepressant drugs act by restoring excitatory synaptic strength. Our findings suggest novel therapeutic targets for this debilitating disease.


The Role of Oxytocin in Parenting and as Augmentative Pharmacotherapy: Critical Issues and Bold Conjectures.
Despite the sometimes heated debate about the validity of human oxytocin studies, experimental oxytocin research with intranasal administration is a growing field with promising preliminary findings. The effects of intranasally administered oxytocin compared to placebo on brain neural activity have been supported in animal studies and in human studies of neural resting state. In several studies, oxytocin sniffs have been shown to lead to down-regulation of amygdala activation in response to infant attachment vocalisations. Meta-analytic evidence shows that oxytocin enhances the salience of (emotional) stimuli, lowers stress and arousal, and elevates empathic concern and tender care, in particular for offspring and in-group members. Less firm evidence points at the amnestic effects of oxytocin. We also note that the average effect sizes of oxytocin experiments are small to modest, and that most studies include a small number of subjects and thus are seriously underpowered, which implies a high risk for publication bias and nonreplicability. Nevertheless, we argue that the power of within-subjects experiments with oxytocin has been underestimated. Much more work is needed, however, to create a firm knowledge base of the neural and behavioural effects of oxytocin. Human oxytocin research is still taking place in the context of discovery, in which bold conjectures are being generated. In the context of justification, these conjectures should subsequently be subjected to stringent attempts at refutations before we jump to theoretical or clinical conclusions. For this context of justification, we propose a multisite multiple replications project on the social stimuli salience enhancing effect of oxytocin. Clinical application of oxytocin is premature. Meta-analytically, the use of oxytocin in clinical groups tends to show only effectiveness in changing symptomatology in individuals with autism spectrum disorders but, even then, it is not yet a validated therapy and its use is premature because safety and long-term side-effects have not been sufficiently studied, in particular in children.


Different glycosylation profiles of cystatin F alter the cytotoxic potential of natural killer cells.
Cystatin F, a cysteine peptidase inhibitor, is a potent modulator of NK cytotoxicity. By inhibiting granule-mediated cytotoxicity pathway, cystatin F induces formation of non-functional NK cell stage, called split-anergy. We show that N-glycosylation determines the localization and cellular function of cystatin F. Cystatin F mostly exhibited high-mannose glycosylation in U-937 cells, both high-mannose and complex glycosylation in NK-92 and primary NKs, and predominantly complex glycosylation in super-charged NKs. Manipulating N-glycosylation with kifunensine increased high-mannose glycosylation of cystatin F and lysosome localisation, which decreased cathepsin C activity and reduced NK cytotoxicity. Mannose-6-phosphate could significantly reduce the internalization of extracellular cystatin F. By comparing NK cells with different cytotoxic potentials, we found that high-mannose cystatin F was strongly associated with lysosomes and cathepsin C in NK-92 cell line. In contrast, in highly cytotoxic super-charged NKs, cystatin F with complex glycosylation was associated with the secretory pathway and less prone to inhibit cathepsin C. Modulating glycosylation to alter cystatin F localisation could increase the cytotoxicity of NK cells, thereby enhancing their therapeutic potential for treating cancer patients.


Repair of Damaged Articular Cartilage: Current Approaches and Future Directions.
Articular hyaline cartilage is extensively hydrated, but it is neither innervated nor vascularized, and its low cell density allows only extremely limited self-renewal. Most clinical and research efforts currently focus on the restoration of cartilage damaged in connection with osteoarthritis or trauma. Here, we discuss current clinical approaches for repairing cartilage, as well as research approaches which are currently developing, and those under translation into clinical practice. We also describe potential future directions in this area, including tissue engineering based on scaffolding and/or stem cells as well as a combination of gene and cell therapy. Particular focus is placed on cell-based approaches and the potential of recently characterized chondro-progenitors; progress with induced pluripotent stem cells is also discussed. In this context, we also consider the ability of different types of stem cell to restore hyaline cartilage and the importance of mimicking the environment in vivo during cell expansion and differentiation into mature chondrocytes.


Antidepressive-like effect of imperatorin from Angelica dahurica in prenatally stressed offspring rats through 5-hydroxytryptamine system.
Adolescence is a time of continued brain maturation, particularly in limbic and cortical regions, which undoubtedly plays a role in the physiological and emotional changes. Prenatally stressed offspring rats were used to investigate the potential antidepressive-like effects of imperatorin (IMP) extracted from the root of radix angelica. After 4 weeks of treatment of IMP, behavioral tests (sucrose-preference test, forced-swimming test, and open-field test) were measured. 5-hydroxytryptamine (5-HT) concentration in the hippocampus and frontal cortex was measured using an enzyme-linked immunosorbent assay. Serotonin transporters (5-HTT) and 5-HT1A receptor (5-HT1AR) mRNA expression in the hippocampus and frontal cortex were also determined by real-time PCR. Administration with IMP (15 and 30 mg/kg/day, intragastrically) for 28 days markedly increased the percentage of sucrose (anhedonia), decreased the immobility time, and increased the number of total crossings, center crossings, rearing, and grooming in the male prenatally stressed offspring. Meanwhile, we found that 5-HT concentration in the hippocampus and frontal cortex was significantly increased in the IMP-treated group. Subsequently, we found significantly decreased 5-HTT and increased 5-HT1AR mRNA expressions in the hippocampus and frontal cortex after IMP treatment in the prenatally stressed male offspring. IMP showed antidepressive-like effects and increased 5-HT concentration in male prenatally stressed offspring, suggesting that IMP could be of therapeutic use in preventing depressive-like behavior in adolescence.


Protein, cell and bacterial fouling resistance of polypeptoid-modified surfaces: effect of side-chain chemistry.
Peptidomimetic polymers consisting of poly-N-substituted glycine oligomers (polypeptoids) conjugated to biomimetic adhesive polypeptides were investigated as antifouling surface coatings. The polymers were immobilized onto TiO(2) surfaces via an anchoring peptide consisting of alternating residues of 3,4-dihydroxyphenylalanine (DOPA) and lysine. Three polypeptoid side-chain compositions were investigated for antifouling performance and stability toward enzymatic degradation. Ellipsometry and XPS analysis confirmed that purified polymers adsorbed strongly to TiO(2) surfaces, and the immobilized polymers were resistant to enzymatic degradation as demonstrated by mass spectrometry. All polypeptoid-modified surfaces exhibited significant reductions in adsorption of lysozyme, fibrinogen and serum proteins, and were resistant to 3T3 fibroblast cell attachment for up to seven days. Long-term in vitro cell attachment studies conducted for six weeks revealed the importance of polypeptoid side-chain composition, with a methoxyethyl side chain providing superior long-term fouling resistance compared to hydroxyethyl and hydroxypropyl side chains. Finally, attachment of both gram-positive and gram-negative bacteria for up to four days under continuous-flow conditions was significantly reduced on the polypeptoid-modified surfaces compared to unmodified TiO(2) surfaces. The results reveal the influence of polypeptoid side-chain chemistry on short-term and long-term protein, cell and bacterial fouling resistance.


Structure and restriction enzyme maps of the circularly permuted DNA of staphylococcal bacteriophage phi 11.
One restriction enzyme map of Staphylococcus aureus bacteriophage phi 11 DNA was established by reciprocal double digestions with the enzymes EcoRI, HaeII, and KpnI. The sequential order of the EcoRI fragments was thereafter established by a novel approach involving blotting of DNA partially cleaved with EcoRI and the probing the blots with nick-translated terminal fragments. A circular map of the phi 11 DNA was established, and the phage genome was circularly permuted based on the failure to end label mature viral DNA, restriction maps of replicating DNA, and finally, homoduplex analysis in the electron microscope. A restriction enzyme map of the prophage form of phi 11 DNA was obtained by analysis of chromosomal DNA from a lysogenic strain.


The neural substrates of probabilistic and intertemporal decision making.
Many important decisions involve outcomes that are either probabilistic or delayed. Based on similarities in decision preferences, models of decision making have postulated that the same psychological processes may underlie decisions involving probabilities (i.e., risky choice) and decisions involving delay (i.e., intertemporal choice). Equivocal behavioral evidence has made this hypothesis difficult to evaluate. However, a combination of functional neuroimaging and behavioral data may allow identification of differences between these forms of decision making. Here, we used functional magnetic resonance imaging (fMRI) to examine brain activation in subjects making a series of choices between pairs of real monetary rewards that differed either in their relative risk or their relative delay. While both sorts of choices evoked activation in brain systems previously implicated in executive control, we observed clear distinctions between these forms of decision making. Notably, choices involving risk evoked greater activation in posterior parietal and lateral prefrontal cortices, whereas choices involving delay evoked greater activation in the posterior cingulate cortex and the striatum. Moreover, activation of regions associated with reward evaluation predicted choices of a more-risky option, whereas activation of control regions predicted choices of more-delayed or less-risky options. These results indicate that there are differences in the patterns of brain activation evoked by risky and intertemporal choices, suggesting that the two domains utilize at least partially distinct sets of cognitive processes.