Combining antiangiogenic therapy with immunotherapy exerts better therapeutical effects on large tumors in a woodchuck hepatoma model.
Cytokine and antiangiogenic gene therapies have proved effective in implanted hepatocellular carcinoma (HCC) models in which small tumor burdens were established in small rodents. These models, however, may not reflect human HCCs, which are frequently detected at a stage when tumors are large and multifocal. In addition, HCC in patients is often associated with viral hepatitis. To investigate the effectiveness of a mixture type of gene therapy strategy on large tumor burdens, we used the woodchuck model in which woodchuck hepatitis virus-induced HCCs are large and multifocal, simulating the conditions in humans. Adenoviruses encoding antiangiogenic factors (pigment epithelium-derived factor and endostatin) or cytokines (GM-CSF and IL-12) were delivered via the hepatic artery separately or in combination into woodchuck livers bearing HCCs. Our results showed that the mixture type of strategy, which contained two cytokines and two antiangiogenic factors, had better antitumor effects on large tumors as compared with monotherapy either with antiangiogenic or cytokine genes. The immunotherapy recruited significant levels of CD3(+) T cells that infiltrated the tumors, whereas the antiangiogenesis-based therapy significantly reduced tumor vasculature. The mixture type of gene therapy achieved both effects. In addition, it induced high levels of natural killer cells and apoptotic cells and reduced the levels of immunosuppressive effectors in the tumor regions. Hence, antiangiogenic therapy may provide the advantage of reducing immune tolerance in large tumors, making them more vulnerable to the immune reactions. Our study implies that in the future, the combination therapy may prove effective for the treatment of patients with advanced HCC.


A Mechanism for Verifying the Integrity and Immutability of Tuberculosis Data Using IOTA Distributed Ledger Technology.
Intensified research and innovation and rapid uptake of new tools, interventions, and strategies are crucial to fight Tuberculosis, the world's deadliest infectious disease. The sharing of health data remains a significant challenge. Data consumers must be able to verify the consistency and integrity of data. Solutions based on distributed ledger technologies may be adequate, where each member in a network holds a unique credential and stores an identical copy of the ledger and contributes to the collective process of validating and certifying digital transactions. This work proposes a mechanism and presents a use case in Digital Health to allow the verification of integrity and immutability of TB electronic health records. IOTA was selected as a supporting tool due to its data immutability, traceability and tamper-proof characteristics. A mechanism to verify the integrity of data through hash functions and the IOTA network is proposed. Then, a set of TB related information systems was integrated with the network. IOTA technology offers performance and flexibility to enable a reliable environment for electronic health records.


Validity of self-reported drug use in chronic pain patients.
Previous researchers have reported that in psychiatric populations many patients provide incorrect self-report information on current drug use. Therefore, the purposes of the present study were to determine the percentage of chronic pain patients (CPPs) using illicit drugs (cannabis, cocaine), to determine the percentage of CPPs who provide incorrect self-report drug use information in the psychiatric examination, and to identify some variables that could help in identifying the CPP likely to provide an incorrect drug use history using drug urine toxicologies. DESIGN/SETTING/PARTICIPANTS/OUTCOME MEASURES: Two hundred seventy-four CPP consecutive admissions to a pain facility were psychiatrically examined according to criteria in the Diagnostic and statistical manual of mental disorders (3rd ed., rev; DSM-III-R), with special emphasis on all current drug use. Immediately after the psychiatric examination, all CPPs were asked to consent to urine toxicology. Urine was tested for benzodiazepines, opioids, tricyclics, propoxyphene, cannabinoids, barbiturates, amphetamines, methadone, methaqualone, phencyclidine, alcohol, and cocaine. CPPs were then segregated into three groups: negative toxicology, positive toxicology but concordant with self-report of current drug use, and positive toxicology discordant with self-report of current drug use. These groups were statistically compared with each other with regard to age, gender, race, workers' compensation status, and prevalence of individual DSM-III-R psychoactive substance use disorders. Sensitivities were also calculated for two conditions: accuracy of toxicology and accuracy of self-report. Toxicologies were obtained from 226 (82.5%) of the CPPs. Toxicologies were negative in 121 (53.5%) and positive in 105 (46.5%) of the CPPs. Of the 226 CPPs, 8.4% had illicit drugs in the urine (6.2% cannabis, 2.2% cocaine). Twenty (8.8%) of the CPPs provided incorrect self-report information about current drug use, the incorrect information most frequently about illicit drugs. Drug urine toxicology sensitivity results indicated that a significant percentage of CPPs was claiming to be taking a drug but was not taking it or taking it incorrectly. The psychiatric examination drug self-report sensitivity results indicated that a significant percentage of CPPs was withholding or providing incorrect information on current drug use. Lowest self-report sensitivity results were in reference to illicit drugs. CPPs who were more likely to provide incorrect psychiatric examination self-report information about current drug use were more likely to be younger, to be a workers' compensation CPP, and to have been assigned a DSM-III-R diagnosis of polysubstance abuse in remission. A significant percentage of CPPs appears to provide incorrect information on current illicit drug use. Urine toxicology studies may have a place in the identification of drugs for which incorrect information may be provided by CPPs. There are many possible reasons, such as assay error, that could lead to apparent misinformation. In the clinical setting, these possibilities should be considered if urine toxicology results appear to be incongruent with psychiatric examination drug use self-report.


[Epidemiologic aspects of the burnout syndrome in hospital personnel].
Burnout Syndrome is claimed to be the most probable cause of the lack of motivation suffered by professional health workers nowadays. This suggests that the syndrome may be linked to the high levels of absenteeism from work among this professional group. The study aims to provide a number of descriptions of the universal epidemiological variables that would allow us to draw up a risk profile for this profession. We studied a random sample of 294 professionals working in the primary specialised health sector to which we applied the Burnout Syndrome measurement instrument (Maslach Burnout Inventory) which was self-administered. Descriptive statistics were gathered with a comparison of average values for socio-demographic variables (P < 0.05) using Epiinfo V.60 and SPSS PC.W. We obtained 87.76% responses compared with 12.23% losses. This sample gave us a 95% reliability level with a 5% error margin. We obtained significant differences in line with sex, age, marital status, length of service in the workplace, number of workers, place of work, number of patients under their responsibility, weekly working hours, patient interaction time. The Burnout average was 47.16 +/- 7.93, with the highest proportions corresponding to emotional fatigue and lack of self-fulfilment. The epidemiological risk profile obtained would be as follows: a female, over 44 years old, with no stable partner, with more than 19 years service in the profession and more than 11 at that particular workplace, working in a specialised department, with more than 21 patients under her responsibility, devoting more than 70% of the working day to these patients and with a working week of 36-40 hours.


Pan-Resistome Characterization of Uropathogenic Escherichia coli and Klebsiella pneumoniae Strains Circulating in Uganda and Kenya, Isolated from 2017-2018.
Urinary tract infection (UTI) develops after a pathogen adheres to the inner lining of the urinary tract. Cases of UTIs are predominantly caused by several Gram-negative bacteria and account for high morbidity in the clinical and community settings. Of greater concern are the strains carrying antimicrobial resistance (AMR)-conferring genes. The gravity of a UTI is also determined by a spectrum of other virulence factors. This study represents a pilot project to investigate the burden of AMR among uropathogens in East Africa. We examined bacterial samples isolated in 2017-2018 from in- and out-patients in Kenya (KY) and Uganda (UG) that presented with clinical symptoms of UTI. We reconstructed the evolutionary history of the strains, investigated their population structure, and performed comparative analysis their pangenome contents. We found 55 Escherichia coli and 19 Klebsiella pneumoniae strains confirmed uropathogenic following screening for the prevalence of UTI virulence genes including fimH, iutA, feoA/B/C, mrkD, and foc. We identified 18 different sequence types in E. coli population while all K. pneumoniae strains belong to ST11. The most prevalent E. coli sequence types were ST131 (26%), ST335/1193 (10%), and ST10 (6%). Diverse plasmid types were observed in both collections such as Incompatibility (IncF/IncH/IncQ1/IncX4) and Col groups. Pangenome analysis of each set revealed a total of 2862 and 3464 genes comprised the core genome of E. coli and K. pneumoniae population, respectively. Among these are acquired AMR determinants including fluoroquinolone resistance-conferring genes aac(3)-Ib-cr and other significant genes: aad, tet, sul1, sul2, and cat, which are associated with aminoglycoside, tetracycline, sulfonamide, and chloramphenicol resistance, respectively. Accessory genomes of both species collections were detected several β-lactamase genes, blaCTX-M, blaTEM and blaOXA, or blaNDM. Overall, 93% are multi-drug resistant in the E. coli collection while 100% of the K. pneumoniae strains contained genes that are associated with resistance to three or more antibiotic classes. Our findings illustrate the abundant acquired resistome and virulome repertoire in uropathogenic E. coli and K. pneumoniae, which are mainly disseminated via clonal and horizontal transfer, circulating in the East African region. We further demonstrate here that routine genomic surveillance is necessary for high-resolution bacterial epidemiology of these important AMR pathogens.


Controlled Attenuation Parameter and Liver Stiffness Measurements for Steatosis Assessment in the Liver Transplant of Brain Dead Donors.
One of the main selection criteria of the quality of a liver graft is the degree of steatosis, which will determine the success of the transplantation. The aim of this study was to evaluate the ability of FibroScan and its related methods Controlled Attenuation Parameter and Liver Stiffness to assess objectively steatosis and fibrosis in livers from brain-dead donors to be potentially used for transplantation. Over a period of 10 months, 23 consecutive brain dead donors screened for liver procurement underwent a FibroScan and a liver biopsy. The different predictive models of liver retrievability using liver biopsy as the gold standard have led to the following area under receiver operating characteristic curve: 76.6% (95% confidence intervals [95% CIs], 48.2%-100%) when based solely on controlled attenuation parameter, 75.0% (95% CIs, 34.3%-100%) when based solely on liver stiffness, and 96.7% (95% CIs, 88.7%-100%) when based on combined indices. Our study suggests that a preoperative selection of brain-dead donors based on a combination of both Controlled Attenuation Parameter and Liver Stiffness obtained with FibroScan could result in a good preoperative prediction of the histological status and degree of steatosis of a potential liver graft.


Review of optical coherence tomography for intraocular tumors.
Optical coherence tomography has assumed an important role in the management of numerous ocular conditions. With regard to ocular oncology, optical coherence tomography can illustrate retinal changes overlying choroidal tumors. Some of these features include photoreceptor loss, intraretinal edema, and retinal thinning overlying choroidal nevus; fresh subretinal fluid with preservation of photoreceptors overlying choroidal melanoma; and intraretinal edema, retinoschisis, and retinal thinning overlying irradiated choroidal melanoma. The optical coherence tomography features of tumors of the retinal pigment epithelium include typical findings of peaked vitreoretinal traction and retinal disorganization with combined hamartoma of the retina and retinal pigment epithelium, full-thickness retinal shadowing with congenital simple hamartoma, and photoreceptor loss and retinal thinning overlying congenital hypertrophy of the retinal pigment epithelium. Optical coherence tomography of retinal tumors, such as retinoblastoma and astrocytic hamartoma, reveals full-thickness replacement of the retinal anatomic layers with the tumor and shadowing corresponding to the intralesional calcification. For all intraocular tumors, optical coherence tomography provides valuable information regarding the status of the retina and the retinal pigment epithelium and can be useful in ascertaining reasons for visual loss.


Sinus histiocytosis with massive lymphadenopathy.
Five cases of "sinus histiocytosis with massive lymphadenopathy" are reported. The clinical features and histopathological appearances are described. The majority were African children with massive cervical lymphadenopathy. Histologically, the lymphnode sinuses are widely dilated by mature histiocytes, with phagocytosis of lymphocytes and other hematopoietic cells by the proliferating histiocytes. The diagnostic difficulties are discussed. These are the first cases recognized in Zambia.


Bovine tuberculosis in a possum from the Orongorongo Valley, Wellington.
The first confirmed case of a tubercular possum in the Orongorongo Valley forest is reported from a population intensively studied for 20 years. The victim was a five-year-old male recaptured 20 times over 3% years. Its annual home range was about 1.8 hectares but this dropped to 0.4 ha during its last year of life. Its range overlapped with at least 15 others. The animal appeared normal and healthy in July 1985 but died of advanced, generalised bovine tuberculosis 12 weeks later. The nearest confirmed cases of tubercular possums were reported 1.3 km away in 1976 and 4 km away in 1979. The Orongorongo possum was unlikely to have contacted any cattle for over three years. The infection was probably transmitted from the Wainuiomata Valley by possums or pigs. The Orongorongo River was no barrier to the spread of the infection. Although most of the neighbouring possums were trapped and examined several times over the following year, no tubercular animals were detected.


Clinical evaluation of herbal coded formulation Cran-off to Urixin in the treatment of urinary tract infection.
Urinary Tract Infections are the largest group of infections after the respiratory tract infections. In 85% of the cases the causative organism is E. Coli. A clinical trial was conducted to evaluate the efficacy of coded herbal formulation "Cranoff" (Test drug) for the treatment of Urinary tract infection comparing with Urixin (Control). One hundred and thirty patients suffering from Urinary tract infection from both groups (Males: 45, mean age: 34±14 and females: 85, mean age: 33±13 year, range: 15-55) were enrolled in the trial and divided in to two groups according to treatment regimens. Cranoff (Test drug) 500mg two capsules and Urixin (Pipemidic Acid Trihydrate JP15) (Control) 400mg capsules twice daily were prescribed for 2-3 weeks. Urinary tract infection was improved in 23 (35.38%) patients out of 65 patients by the use of Cranoff (Test drug), and in 15 (23.07%) patients out of 65 by the use of Urixin (Control drug). Furthermore, there was a significant improvement in Urinary tract infection associated clinical features as compared to Urixin. It is concluded that Cranoff possesses a therapeutic value for the improvement of urinary tract infection and its associated symptoms as compared to Urixin.


Affective touch awareness in mental health and disease relates to autistic traits - An explorative neurophysiological investigation.
Affective touch is important for social interaction within families and groups and there is evidence that unmyelinated C tactile fibers are involved in this process. Individuals with autism spectrum disorders show alterations in the perception and processing of affective touch. sThus, we hypothesized that affective touch awareness based on C tactile fiber activation is impaired in individuals with high levels of autistic trait. The pleasantness perception of optimal and suboptimal C tactile stimuli was tested in an explorative study in 70 patients recruited from an outpatient psychotherapy clinic and 69 healthy comparison subjects. All participants completed questionnaires about autistic traits, depressive symptomatology, childhood maltreatment, and about the daily amount of touch. Relative to comparison subjects, patients reported engaging in touch less frequently in daily life and rated touch less pleasant. Reduced valence ratings of touch were explained by childhood maltreatment but not by any particular disorder or depression severity. Among all tested variables, the affective touch awareness correlated with autistic traits only - in patients as well as in comparison subjects. Taken together, individuals with mental health issues have a lower baseline of expression and reception of affective touch. Autistic traits and childhood maltreatment modulate the experience of affective touch.


Comparison of the rotation-advancement and philtral ridge techniques for unilateral cleft lip repair.
The Millard rotation-advancement flap has become the most widely used technique for unilateral cleft lip repair. The philtral ridge repair is a modified straight-line technique that was developed to further optimize the scar associated with the rotation-advancement flap. The purpose of this article is to introduce the philtral ridge repair and objectively compare the outcomes of these two techniques. Two senior board-certified surgeons, who are active members of their respective craniofacial teams, use different surgical techniques for the unilateral cleft lip: the philtral ridge and rotation-advancement repairs. The authors retrospectively analyzed preoperative and postoperative photographs of consecutive patients who underwent repair performed by each surgeon between 2003 and 2009. Using Adobe Photoshop imaging software, facial points on the cleft and noncleft sides were measured, including height and symmetry of Cupid's bow, width and height of the nasal vestibule, height of the vermilion, and alar base position. Ratios of cleft side to noncleft side measurements were calculated to standardize comparisons between patients. In addition, the symmetry of each lip repair was graded subjectively by health care professionals and the general public. There were no differences in preoperative ratios between the two techniques with the exception of a wider cleft nasal vestibule in the rotation-advancement group (p = 0.04). There were no statistically significant differences in postoperative measures or subjective analysis of symmetry between the groups. Both the rotation-advancement and philtral ridge techniques produced outcomes with a high degree of facial symmetry and are excellent options for unilateral cleft lip repair.


The Impact of Lipid Corona on Rifampicin Intramacrophagic Transport Using Inhaled Solid Lipid Nanoparticles Surface-Decorated with a Mannosylated Surfactant.
The mimicking of physiological conditions is crucial for the success of accurate in vitro studies. For inhaled nanoparticles, which are designed for being deposited on alveolar epithelium and taken up by macrophages, it is relevant to investigate the interactions with pulmonary surfactant lining alveoli. As a matter of fact, the formation of a lipid corona layer around the nanoparticles could modulate the cell internalization and the fate of the transported drugs. Based on this concept, the present research focused on the interactions between pulmonary surfactant and Solid Lipid Nanoparticle assemblies (SLNas), loaded with rifampicin, an anti-tuberculosis drug. SLNas were functionalized with a synthesized mannosylated surfactant, both alone and in a blend with sodium taurocholate, to achieve an active targeting to mannose receptors present on alveolar macrophages (AM). Physico-chemical properties of the mannosylated SLNas satisfied the requirements relative to suitable respirability, drug payload, and AM active targeting. Our studies have shown that a lipid corona is formed around SLNas in the presence of Curosurf, a commercial substitute of the natural pulmonary surfactant. The lipid corona promoted an additional resistance to the drug diffusion for SLNas functionalized with the mannosylated surfactant and this improved drug retention within SLNas before AM phagocytosis takes place. Moreover, lipid corona formation did not modify the role of nanoparticle mannosylation towards the specific receptors on MH-S cell membrane.


Using a slightly tapered optical fiber to attract and transport microparticles.
We exploit a fiber puller to transform a telecom single-mode optical fiber with a 125 microm diameter into a symmetric and unbroken slightly tapered optical fiber with a 50 microm diameter at the minimum waist. When the laser light is launched into the optical fiber, we can observe that, due to the evanescent wave of the slightly tapered fiber, the nearby polystyrene microparticles with 10 microm diameters will be attracted onto the fiber surface and roll separately in the direction of light propagation. We have also simulated and compared the optical propulsion effects on the microparticles when the laser light is launched into a slightly tapered fiber and a heavily tapered (subwavelength) fiber, respectively.


Photobiomodulation Activates Microglia/Astrocytes and Relieves Neuropathic Pain in Inferior Alveolar Nerve Injury.
Objective: This study aimed to determine microglial/astrocyte changes and their associated analgesic effect in inferior alveolar nerve injury (IANI) model rats treated with photobiomodulation therapy (PBMT) using a 940-nm diode laser. Background: Very few basic studies have investigated microglial/astrocyte dynamics following PBMT aimed at relieving neuropathic pain caused by IANI. Methods: Rats were divided into an IANI-PBM group, IANI+PBM group, and sham+PBM group. Observations were made on the day before IANI or the sham operation and on postoperative days 3, 5, 7, 14, and 28. PBMT was delivered for 7 consecutive days, with an energy density of 8 J/cm2. Behavioral analysis was performed to determine pain thresholds, and immunohistological staining was performed for the microglia marker Iba1 and astrocyte marker glial fibrillary acidic protein, which are observed in the spinal trigeminal nucleus. Results: Behavioral analysis showed that the pain threshold returned to the preoperative level on postoperative day 14 in the IANI+PBM group, but decreased starting from postoperative day 1 and did not improve thereafter in the IANI-PBM group (p ≤ 0.001). Immunological analysis showed that microglial and astrocyte cell counts were similar in the IANI+PBM group and IANI-PBM group shortly after IANI (day 3), but the expression area was larger (p ≤ 0.001) and hypertrophy of microglia and astrocyte cell bodies and end-feet extension (i.e., indicators of activation) were more prominent in the IANI+PBM group. Conclusions: PBMT after IANI prevented hyperalgesia and allodynia by promoting glial cell activation shortly after injury.


Explaining the Variable Effects of Social Support on Work-Based Stressor-Strain Relations: The Role of Perceived Pattern of Support Exchange.
Seeking to explain mixed empirical findings regarding the buffering effect of social support on work-based stress-strain relations, we posit that whether an increase in the level of support received buffers or exacerbates the harmful effects of workload on employee health and well-being is contingent upon the general pattern characterizing an employee supportive exchanges across his/her close relationships. Specifically, we propose that the buffering effect of receiving social support depends on whether the employee perceives his/her social exchanges as reciprocal (support given equals support received), under-reciprocating (support given exceeds support received), or over-reciprocating (support received exceeds support given). Based on longitudinal data collected from a random sample of blue-collar workers, our findings support our predictions, indicating that the buffering effect of social support on the relationship between work hours (on the one hand) and employee health and well-being (on the other) varies as a function of the pattern of exchange relations between an employee and his/her close support providers.


Global field synchronization in gamma range of the sleep EEG tracks sleep depth: Artifact introduced by a rectangular analysis window.
Investigating functional connectivity between brain networks has become an area of interest in neuroscience. Several methods for investigating connectivity have recently been developed, however, these techniques need to be applied with care. We demonstrate that global field synchronization (GFS), a global measure of phase alignment in the EEG as a function of frequency, must be applied considering signal processing principles in order to yield valid results. Multichannel EEG (27 derivations) was analyzed for GFS based on the complex spectrum derived by the fast Fourier transform (FFT). We examined the effect of window functions on GFS, in particular of non-rectangular windows. Applying a rectangular window when calculating the FFT revealed high GFS values for high frequencies (>15Hz) that were highly correlated (r=0.9) with spectral power in the lower frequency range (0.75-4.5Hz) and tracked the depth of sleep. This turned out to be spurious synchronization. With a non-rectangular window (Tukey or Hanning window) these high frequency synchronization vanished. Both, GFS and power density spectra significantly differed for rectangular and non-rectangular windows. Previous papers using GFS typically did not specify the applied window and may have used a rectangular window function. However, the demonstrated impact of the window function raises the question of the validity of some previous findings at higher frequencies. We demonstrated that it is crucial to apply an appropriate window function for determining synchronization measures based on a spectral approach to avoid spurious synchronization in the beta/gamma range.


Review of current clinical biomarkers for the detection of alcohol dependence.
Alcohol dependence is often seen in a variety of clinical settings and requires attention to reduce medical complications, set up appropriate treatments, and minimize utilization of healthcare resources. Patient responses to questionnaires are often used to screen for alcohol problems, but can be misleading in the context of altered mental states or in a patient hesitant to disclose a pattern of alcohol use. Identifying the biochemical consequences of alcohol dependence has led to further study, including correlating laboratory findings to increase accuracy of identifying problem drinkers. Understanding the normal function, mechanism of abnormal findings, sensitivity, and specificity of the current laboratory studies can substantiate clinical suspicion of alcohol use. In this article, we provide results from our literature search regarding laboratory abnormalities in alcohol dependence, review options available to complement a thorough history and physical, and provide a brief overview of future biomarkers for detection of alcohol use.


Optimal oxygen saturation for preterm babies. Do we really know?
Oxygen is the most commonly used 'drug' in neonatal units as an integral part of respiratory support. It has also been known for half of the century that it is easy to damage the eyes of preterm infants by giving too much oxygen especially in the first few weeks of life. Despite this knowledge there is still a wide variation in approaches to oxygen monitoring within neonatal units. A randomized controlled trial conducted more than 50 years ago first made clinicians aware of 'oxygen toxicity' in preterm infants, but no other controlled trial has ever been conducted since to clarify how much oxygen infants really need, or what oxygen saturation level is optimal in caring these preterm babies. Perhaps time has come for clinicians to resolve this 'uncertainty' by well-designed randomized trials.


Defining the incidence of cardiorespiratory instability in patients in step-down units using an electronic integrated monitoring system.
To our knowledge, detection of cardiorespiratory instability using noninvasive monitoring via electronic integrated monitoring systems (IMSs) in intermediate or step-down units (SDUs) has not been described. We undertook this study to characterize respiratory status in an SDU population, to define features of cardiorespiratory instability, and to evaluate an IMS index value that should trigger medical emergency team (MET) activation. This descriptive, prospective, single-blinded, observational study evaluated all patients in a 24-bed SDU in a university medical center during 8 weeks from November 16, 2006, to January 11, 2007. An IMS (BioSign; OBS Medical, Carmel, Indiana) was inserted into the standard noninvasive hardwired monitoring system and used heart rate, blood pressure, respiratory rate, and peripheral oxygen saturation by pulse oximetry to develop a single neural networked signal, or BioSign Index (BSI). Data were analyzed for cardiorespiratory instability according to BSI trigger value and local MET activation criteria. Staff were blinded to BSI data collected in 326 patients (total census). Data for 18 248 hours of continuous monitoring were captured. Data for peripheral oxygen saturation by pulse oximetry were absent in 30% of monitored hours despite being a standard of care. Cardiorespiratory status in most patients (243 of 326 [74.5%]) was stable throughout their SDU stay, and instability in the remaining patients (83 of 326 [25%]) was exhibited infrequently. We recorded 111 MET activation criteria events caused by cardiorespiratory instability in 59 patients, but MET activation for this cause occurred in only 7 patients. All MET events were detected by BSI in advance (mean, 6.3 hours) in a bimodal distribution (>6 hours and < or =45 minutes). Cardiorespiratory instability, while uncommon and often unrecognized, was preceded by elevation of the IMS index. Continuous noninvasive monitoring augmented by IMS provides sensitive detection of early instability in patients in SDUs.


Neuronal Autophagy in Synaptic Functions and Psychiatric Disorders.
Homeostatic maintenance of physiological functions is fundamental to organismal well-being. Disruption or imbalance in homeostasis results in functional disturbances at molecular, cellular, and tissue levels, leading to manifestation as physical and mental illnesses. Homeostatic imbalance is caused by a range of pathophysiological mechanisms, including disrupted reduction-oxidation reactions, inflammatory responses, metabolic disturbances, or failure in quality control of cellular proteins and organelles. However, the roles for the protein/organelle quality control in the regulation of behaviors, in particular of cognitive processes, had not been well documented, until recent reports finally supported this concept. The frontline studies in neuroscience have revealed that synaptic components (e.g., synaptic proteins, organelles, neurotransmitters and their receptors) are selectively degraded by autophagy, a cellular recycling machinery implicated in surveillance and quality control of proteins and organelles responsible for the maintenance of cellular homeostasis. Apart from the canonical role of autophagy in supporting cell viability, synaptic autophagy appears to regulate synapse remodeling and plasticity. Consistently, emerging evidence suggests novel roles of autophagy in memory encoding, information processing, or cognitive functions. In this review, we overview recent progress in understanding the roles of neuronal autophagy in homeostatic maintenance of synaptic functions, with particular focus on how disruptions in these processes may contribute to the pathophysiology of psychiatric disorders.


Microchip HPLC of peptides and proteins.
Rapid microchip reversed-phase HPLC of peptides and proteins at pressure gradients of 12 bar/cm (180 psi/cm) has been performed using a microdevice that integrates subnanoliter on-chip injection and separation with a miniaturized fluorescence detector. Proteins and peptides were separated on a C18 side-chain porous polymer monolith defined by contact lithography, and injection was achieved via a pressure-switchable fluoropolymer valve defined using projection lithography. Preliminary separations of peptide standards and protein mixtures were performed in 40-200 s, and switching between samples with no detectible sample carryover has been performed. The injections and separations were reproducible; the relative standard deviation (RSD) for retention time was 0.03%, and peak area RSD was 3.8%. Sample volumes ranging from 220 to 800 pL could be linearly metered by controlling the pressure injection pulse duration with conventional timing and valving. The current prototype system shows the potential for rapid and autonomous HPLC separations with varying modalities and the potential for direct connection to mass spectrometers at nanospray flow rates.


Preoperative Japanese Society for the Surgery of the Foot Lesser toe score and erythrocyte sedimentation rate influence wound healing following rheumatoid forefoot surgery.
Delayed wound healing is one of the most common complications following forefoot surgery in patients with rheumatoid arthritis. We aimed to identify the risk factors for delayed wound healing following rheumatoid forefoot surgery. Consecutive patients who underwent primary rheumatoid forefoot surgery (86 feet; 53 patients) between April 2008 and February 2019 were retrospectively evaluated. Clinical data, including smoking history, duration of the disease, presence of diabetes mellitus, medication, white blood cell count, erythrocyte sedimentation rate (ESR), C-reactive protein, the surgical procedure performed, and the Japanese Society for Surgery of the Foot (JSSF) scores, were collected. Delayed wound healing was identified in 20 of 86 (23.3%) feet. In univariate analysis, participants showing delayed healing were older at the time of surgery (p = .04), their ESR was higher (p = .0006), and their total (p = .019) and pain (p = .016) scores on the JSSF Lesser toe scale were lower than those showing normal healing. In multivariable analysis, both the total preoperative JSSF Lesser toe scale score (p = .0239) and ESR (p = .0126) remained significant risk factors for delayed wound healing. After rheumatoid forefoot surgery, surgeons should pay more attention to wound care in patients with lower preoperative JSSF Lesser toe score and high ESR.


Kyleena - another levonorgestrel IUS.
A levonorgestrel (LNG) intrauterine system (IUS) known as Kyleena (Bayer PLC) received marketing authorisation from the European Medicines Agency in 2016.1 The device contains 19.5mg LNG and is licensed for contraception for up to 5 years. This is the fourth LNG IUS product to be licensed for use in the UK. The company states that Kyleena offers advantages over Mirena and Jaydess LNG IUS devices.2 Here we consider the evidence for Kyleena.


Hydroxysteroid dehydrogenases in the kidney of white-breasted water hen, Amaurornis phoenicurus chinensis (Boddaert).
Histochemical localization of delta5-3beta-hydroxysteroid dehydrogenase (delta5-3beta-HSDH), 17beta-hydroxysteroid dehydrogenase (17beta-HSDH), 11beta-hydroxysteroid dehydrogenase (11beta-HSDH) and glucose-6-phosphate dehydrogenase (G-6-PDH) have been studied in the kidney of white-breasted water hen, Amaurornis phoenicurus chinensis. All these enzyme activities occurred in the proximal and distal convoluted and collecting tubules, however, the intensity of these enzyme activities was more in the proximal convoluted tubules. It is suggested that these enzymes might have a role in converting certain hydroxysteroids to ketosteroids during steroid excretion.


Noninvasive assessment of hepatic fibrosis and necroinflammatory activity in Egyptian children with chronic hepatitis C virus infection using FibroTest and ActiTest.
Hepatitis C virus (HCV) is a major cause of chronic hepatitis and hepatic fibrosis. Liver biopsy, because of its limitations and risks, might be considered an imperfect gold standard for assessing the severity of chronic liver diseases. In this study, we aimed to prospectively validate FibroTest (FT) and ActiTest (AT) as noninvasive serum biochemical markers for assessment of the degree of hepatic fibrosis and necroinflammatory activity respectively, in pediatric patients with chronic HCV infection and compare them to liver biopsy. Fifty patients, aged 2 to 18 years, with chronic HCV infection were prospectively enrolled. Two assessments were carried out, within 24-h duration, one of a liver biopsy specimen and the other FT and AT measured in serum sample. A highly significant linear trend and correlation were found between FT-related fibrosis and fibrosis stage by METAVIR scoring on histopathological examination. A highly significant correlation was also found between AT and necroinflammatory histological activity using METAVIR as well. The FT area under the receiver operating characteristic curve (AUROC) is 0.97, SE=0.02 which can diagnose patients with mild stage of fibrosis, thus discriminating them from those with no (or minimal) fibrosis. The AT can successfully discriminate between patients with moderate activity and those with mild activity with AUROC=0.93, SE=0.06. FT and AT are potential noninvasive methods for assessment of hepatic fibrosis and necroinflammatory activity in pediatric patients with chronic HCV infection in comparison with liver biopsy.


Hypothalamic prolyl endopeptidase (PREP) regulates pancreatic insulin and glucagon secretion in mice.
Prolyl endopeptidase (PREP) has been implicated in neuronal functions. Here we report that hypothalamic PREP is predominantly expressed in the ventromedial nucleus (VMH), where it regulates glucose-induced neuronal activation. PREP knockdown mice (Prep(gt/gt)) exhibited glucose intolerance, decreased fasting insulin, increased fasting glucagon levels, and reduced glucose-induced insulin secretion compared with wild-type controls. Consistent with this, central infusion of a specific PREP inhibitor, S17092, impaired glucose tolerance and decreased insulin levels in wild-type mice. Arguing further for a central mode of action of PREP, isolated pancreatic islets showed no difference in glucose-induced insulin release between Prep(gt/gt) and wild-type mice. Furthermore, hyperinsulinemic euglycemic clamp studies showed no difference between Prep(gt/gt) and wild-type control mice. Central PREP regulation of insulin and glucagon secretion appears to be mediated by the autonomic nervous system because Prep(gt/gt) mice have elevated sympathetic outflow and norepinephrine levels in the pancreas, and propranolol treatment reversed glucose intolerance in these mice. Finally, re-expression of PREP by bilateral VMH injection of adeno-associated virus-PREP reversed the glucose-intolerant phenotype of the Prep(gt/gt) mice. Taken together, our results unmask a previously unknown player in central regulation of glucose metabolism and pancreatic function.


Care continuity impacts medicare expenditures of older adults: Fact or fiction?
Older adults with cardiometabolic conditions are typically seen by multiple providers. Management by multiple providers may compromise care continuity and increase health expenditures for older adults, which may partly explain the inverse association between continuity and Medicare expenditures found in prior studies. This study sought to examine whether all-cause admission, outpatient expenditures or total expenditures were associated with the number of prescribers of cardiometabolic medications. Medicare fee-for-service beneficiaries with diabetes (n = 100,191), hypertension (n = 299,949) or dyslipidemia (n = 243,598) living in 10 states were identified from claims data. The probability of an all-cause hospital admission in 2011 was estimated via logistic regression and Medicare (outpatient, total) expenditures in 2011 were estimated using generalized linear models, both as a function of the number of prescribers in 2010. Regressions were adjusted for demographic characteristics, Medicaid status, number of prescriptions, and 17 chronic conditions. In all three cohorts, older adults with more prescribers in 2010 had modestly higher adjusted odds of all-cause inpatient admission than older adults with a single prescriber. Compared to a single prescriber, outpatient and total expenditures in 2011 were 3-10% higher for older adults with diabetes and multiple prescribers, 2-6% higher for older adults with hypertension and multiple prescribers, and 2-5% higher for older adults with dyslipidemia and multiple prescribers. These results provide some evidence that older adults with multiple prescribers also have modestly higher Medicare utilization than those with a single prescriber; thus care continuity may impact patient utilization. Level III (retrospective cohort analysis).


An image representation algorithm compatible with neural-associative-processor-based hardware recognition systems.
A robust image representation algorithm compatible with the VLSI-matching-engine-based image recognition system has been developed. The spatial distributions of four-principal-direction edges in a 64 /spl times/ 64-pels gray scale image are coded to form a 64-dimension feature vector. Since the 2D edge information is reduced to a feature vector by projecting edge flags to the principal directions, it is named the projected principal-edge distribution (PPED) representation. The PPED vectors very well preserve the human perception of similarity among images in the vector space, while achieving a substantial dimensionality reduction in the image data. The PPED algorithm has been applied to medical radiograph analysis, which was taken as a test vehicle for algorithm optimization. The robust nature of the PPED representation has been confirmed by the recognition results comparable to the diagnosis by experts having several years of experience in a university hospital. Dedicated digital VLSI circuits have been developed for PPED vector generation in order to expedite the processing. A test hardware recognition system was constructed using the vector generation circuits, where the analog neural associative processor chip developed in a separate project was employed as a vector-matching engine. As a result, a successful medical radiograph analysis has been experimentally demonstrated using the hardware system. Feasibility of a very low-power operation of the system has been also demonstrated.


[Gastric mucosa islands in the cervical esophagus. Their potential pathogenetic significance].
Islands of gastric mucosa were sought in non-selected in- and out-patients undergoing endoscopy. Mucosal islands were found in 40 patients (out of approximately 1000 oesopho-gastroduodenoscopies), mostly a few centimetres below the mouth of the oesophagus. Histology showed "pure" corpus mucosa in 24 cases, a combination of fundus glands (with zymogenic and parietal cells) and mucoid glands were shown in 9 cases; the rest had only mucoid glands (3 cases) or surface elements of gastric mucosa (two cases). In two cases the islands were missed in the biopsy. In one case the demonstration of HCl secretion from a corpus mucosal island could be achieved by local application of congo red via endoscope after stimulation with pentagastrin. The pathogenetic significance of HCl-producing gastric mucosal islands in the cervical oesophagus is due to their ability to induce peptic lesions with subsequent web or membrane formation.


Control of root-knot nematodes on tomato in stone wool substrate with biological nematicides.
The efficacy of four biological nematicides on root-galling, root-knot nematode (Meloidogyne incognita) reproduction, and shoot weight of tomato (Solanum lycopersicum) grown in stone wool substrate or in pots with sandy soil was compared to an oxamyl treatment and a non-treated control. In stone wool grown tomato, Avid® (a.i. abamectin) was highly effective when applied as a drench at time of nematode inoculation. It strongly reduced root-galling and nematode reproduction, and prevented a reduction in tomato shoot weight. However, applying the product one week before, or two weeks after nematode inoculation was largely ineffective. This shows that Avid® has short-lived, non-systemic activity. The effects of Avid® on nematode symptoms and reproduction on soil-grown tomato were only very minor, probably due to the known strong adsorption of the active ingredient abamectin to soil particles. The neem derived product Ornazin® strongly reduced tomato root-galling and nematode reproduction only in stone wool and only when applied as a drench one week prior to nematode inoculation, suggesting a local systemic activity or modification of the root system, rendering them less suitable host for the nematodes. This application however also had some phytotoxic effect, reducing tomato shoot weights. The other two products, Nema-Q™ and DiTera®, did not result in strong or consistent effects on nematode symptoms or reproduction.


Discrimination among reduced folates and methotrexate as transport substrates by a phenylalanine substitution for serine within the predicted eighth transmembrane domain of the reduced folate carrier.
A phenylalanine substitution for serine in the reduced folate carrier at residue 309 (RFC1-S309F) was identified in a methotrexate (MTX)-resistant cell line selected with 5-formyltetrahydrofolate (5-CHO-THF) as the sole folate source. The transport characteristics of the mutated carrier were studied by transfection into the MTXrA line, which lacks endogenous RFC1 function. The level of expression of carrier in the cell lines studied was determined by specific surface binding of 5-methyltetrahydrofolate (5-CH3-THF). Influx of 5-CH3-THF and 5-CHO-THF mediated by RFC1-S309F was 20- and 7-fold greater than that of MTX, respectively. Consistent with the influx difference between 5-CHO-THF and MTX, the growth requirement (EC50) for 5-CHO-THF in MTXrA-S309F cells was decreased by a factor of 9, while the MTX IC50 was reduced by a factor of only approximately 2 as compared with the recipient MTXrA cells. The decrease in 5-CH3-THF influx mediated by the mutated carrier was attributed to a decrease in the mobility of the 5-CH3-THF-carrier complex, since the influx Kt was essentially unchanged. However, the reduction in 5-CHO-THF and MTX influx was attributed to decreases in both carrier affinity and Vmax, although the decline in the MTX influx Vmax appeared to be much greater than for 5-CHO-THF. The inhibitory effect of chloride on 5-CHO-THF influx observed for L1210 cells was eliminated in the MTXrA-S309F line. This study represents another example of a single mutation in RFC1 that markedly impairs MTX influx but partially preserves transport of reduced folates when cells are selected with 5-CHO-THF as the available folate substrate. The data indicate that residues in the predicted eighth transmembrane domain of RFC1 can play an important role in the selectivity of folate binding and the mobility of the carrier-substrate complex.


A Codeveloped Web-Based Disability Disclosure Toolkit for Youth With Disabilities: Mixed Methods Pilot Evaluation.
Youth and young adults with disabilities experience many barriers in securing employment such as discrimination, inaccessible environments, and lack of support. Youth often need to decide whether and how they should disclose their need for accommodations to employers, which can help them to do their best at work. However, few evidence-based toolkits focusing on disability disclosure exist for youth with various types of disabilities. Supporting youth to develop self-advocacy skills is salient because they are an underrepresented and marginalized group in the labor market. The objective of this study was to conduct a pilot evaluation of a web-based toolkit to enhance disability disclosure for youth and young adults helping to advocate for their needs and request workplace accommodations. We conducted 2 in-person focus groups to codevelop a web-based disability disclosure toolkit, which was followed by a pilot evaluation with a pre-post survey. Primary outcomes focused on the relevance of the toolkit content, preliminary perceived impact on knowledge and confidence, and open-ended feedback on the usefulness of the toolkit. Secondary outcomes focused on effectiveness (ie, measures of self-determination). A total of 14 youths with various types of disabilities took part in the study (aged 20-25 years; n=11, 78% female) including 3 who participated in the codevelopment focus group sessions and 11 youths who participated in the surveys. Our findings involved three main themes in the codevelopment sessions that included (1) disability disclosure and workplace accommodation experiences (ie, knowing when, whether, and how to disclose their disability and request workplace accommodations), (2) usefulness of the tool (ie, relatable content, format and design, and suggestions for further development), and (3) perceived impact of the toolkit (ie, navigating disclosure decisions and how to approach employers and develop other relevant employment skills). The survey findings showed that the majority of participants (10/11, 91%) reported that the toolkit increased or changed their knowledge or understanding of disability disclosure. Most participants (8/11, 73%) reported that the toolkit helped to increase their perceived confidence in their daily activities. The majority of participants (8/11, 73%) agreed or strongly agreed that the toolkit was easy to understand and comprehensive. Regarding the preliminary impact of the toolkit, participants did not demonstrate any significant improvements in self-determination (all P>.05). Our findings emphasize the importance of codeveloping a disability disclosure toolkit with youth to enhance its relevance for their needs. Our toolkit indicates preliminary potential as an educational resource for youth and young adults with disabilities as they search for and secure employment. Further research is needed to assess the impact of the tool with larger samples to understand the impact of workplace disability disclosure decisions for youth with disabilities.


Discrimination in the United States: Experiences of lesbian, gay, bisexual, transgender, and queer Americans.
To examine reported experiences of discrimination against lesbian, gay, bisexual, transgender, and queer (LGBTQ) adults in the United States, which broadly contribute to poor health outcomes. Data came from a national, probability-based telephone survey of US adults, including 489 LGBTQ adults (282 non-Hispanic whites and 201 racial/ethnic minorities), conducted January-April 2017. We calculated the percentages of LGBTQ adults reporting experiences of discrimination in health care and several other domains related to their sexual orientation and, for transgender adults, gender identity. We report these results overall, by race/ethnicity, and among transgender adults only. We used multivariable models to estimate adjusted odds of discrimination between racial/ethnic minority and white LGBTQ respondents. Experiences of interpersonal discrimination were common for LGBTQ adults, including slurs (57 percent), microaggressions (53 percent), sexual harassment (51 percent), violence (51 percent), and harassment regarding bathroom use (34 percent). More than one in six LGBTQ adults also reported avoiding health care due to anticipated discrimination (18 percent), including 22 percent of transgender adults, while 16 percent of LGBTQ adults reported discrimination in health care encounters. LGBTQ racial/ethnic minorities had statistically significantly higher odds than whites in reporting discrimination based on their LGBTQ identity when applying for jobs, when trying to vote or participate in politics, and interacting with the legal system CONCLUSIONS: Discrimination is widely experienced by LGBTQ adults across health care and other domains, especially among racial/ethnic minorities. Policy and programmatic efforts are needed to reduce these negative experiences and their health impact on sexual and/or gender minority adults, particularly those who experience compounded forms of discrimination.


The role of estradiol metabolism in urogenital schistosomiasis-induced bladder cancer.
Urogenital schistosomiasis is a neglected tropical disease that can lead to bladder cancer. How urogenital schistosomiasis induces carcinogenesis remains unclear, although there is evidence that the human blood fluke Schistosoma haematobium, the infectious agent of urogenital schistosomiasis, releases estradiol-like metabolites. These kind of compounds have been implicated in other cancers. Aiming for enhanced understanding of the pathogenesis of the urogenital schistosomiasis-induced bladder cancer, here we review, interpret, and discuss findings of estradiol-like metabolites detected in both the parasite and in the human urine during urogenital schistosomiasis. Moreover, we predict pathways and enzymes that are involved in the production of these metabolites emphasizing their potential effects on the dysregulation of the tumor suppressor gene p53 expression during urogenital schistosomiasis. Enhanced understanding of these potential carcinogens may not only shed light on urogenital schistosomiasis-induced neoplasia of the bladder, but would also facilitate development of interventions and biomarkers for this and other infection-associated cancers at large.


Na+-K+ regulation in cultured vascular smooth muscle cell of the spontaneously hypertensive rat.
Na+-K+ passive transport and activity of the Na+ pump were examined in serially passed cultured vascular smooth muscle cells originating from spontaneously hypertensive (SH), Wistar-Kyoto (WKY), and Wistar (W) rats. Measurements included 22Na+ and 86Rb+ (K+ analogue) uptake and washout rate constants as well as intracellular Na+ and K+ levels. The aforementioned variables were studied in cells subjected to either 2 mM Ca2+ or Ca2+-deficient media. In 2 mM Ca2+ medium, SH rat cells demonstrated the highest exchange (uptake and washout) rate constants for Na+ and Rb+ (K+) among cells of the three rat strains. At this extracellular Ca2+ concentration, the Na+ pump activity of SH rat cells was higher than that of WKY rat cells and was not different from that of W rat cells. Incubation in Ca2+-deficient medium resulted in increased magnitudes of Rb+ washout and Na+ uptake rate constants in all cell preparations associated with elevated intracellular Na+ concentrations and augmented activity of the Na+ pump. Under this condition, cells derived from SH rats showed the highest Na+ uptake and Rb+ washout rate constants associated with the highest Na+ pump activity. The increase in intracellular Na+ level in Ca2+-deficient medium was the highest in SH rat cells. These findings show that innate membrane defects and the response of the Na+ pump to these abnormalities can be demonstrated in in vitro-grown vascular smooth muscle cells of the SH rat.


[Certificate education for geriatric physician: satisfaction and feasibility].
Korea faces a number of challenges to meet demands in the area of geriatric professional medicine in a country with a rapidly ageing population. We evaluated the satisfaction and feasibility of the current education certification for geriatric physicians. Geriatric physicians who were deemed qualified by the Korean Geriatrics Society during the period of 2001 to 2005 (n=2,200) were asked to complete structured questionnaires sent to them by mail about their satisfaction of and need for certificates of education, as well as their opinions on their geriatric specialty training. A total of 419 physicians responded. Descriptive analysis and hierarchical regression were performed to rate the respondents.satisfaction, the characteristics of the need for clarity and utility in education certification, and the characteristics of their patients. Although most respondents were satisfied with their education certification, those who had more elderly patients, aged 65 or older, and those who had more cognitively impaired patients, rated their education as significantly lower than did other physicians. Both groups expressed the need for more the comprehensive care and assessment concerning of their education. Multiple regression analysis indicated that satisfaction with geriatric physician qualification was associated with a physician's age, specialty, and percentage of elderly patients. This study suggests that the current system of education certification is limited in terms of feasibility and physician satisfaction.


Optical control of AMPA receptors using a photoswitchable quinoxaline-2,3-dione antagonist.
AMPA receptors respond to the neurotransmitter glutamate and play a critical role in excitatory neurotransmission. They have been implicated in several psychiatric disorders and have rich pharmacology. Antagonists of AMPA receptors have been explored as drugs and one has even reached the clinic. We now introduce a freely diffusible photoswitchable antagonist that is selective for AMPA receptors and endows them with light-sensitivity. Our photoswitch, ShuBQX-3, is active in its dark-adapted trans-isoform but is significantly less active as its cis-isoform. ShuBQX-3 exhibits a remarkable red-shifting of its photoswitching properties through interactions with the AMPA receptor ligand binding site. Since it can be used to control action potential firing with light, it could emerge as a powerful tool for studying synaptic transmission with high spatial and temporal precision.


Effect of maysin on wild-type, deltamethrin-resistant, and Bt-resistant Helicoverpa armigera (Lepidoptera: Noctuidae).
Larvae of the Old World corn earworm, Helicoverpa armigera (Hübner), were fed diets containing lyophilized silks from maize genotypes expressing varying levels of maysin, a flavone glycoside known to be toxic to the New World corn earworm, Helicoverpa zea Boddie. Three different H. armigera colonies were tested: a wild-type colony (96-S), a colony selected for resistance to deltamethrin (Del-R), and a colony selected for resistance to the Cry1Ac protoxin of Bacillus thuringiensis (Bt-R). A colony of H. zea was also tested as a control. High-maysin silk diets significantly slowed the growth and arrested the development of larvae from all H. armigera colonies compared with low-maysin silk diets, maysin-lacking silk diets, and no-silk control diets. The effects on the H. armigera and H. zea colonies were similar across maysin levels, although H. zea is a larger insect than H. armigera and this overall size difference was observed. Among the H. armigera colonies, maysin effects were generally similar, although 7-d-old Del-R larvae were significantly smaller than 7-d-old Bt-R and 96-S larvae for one no-silk control and two maysin-containing silk treatments. The toxic effect of maysin on the Bt-R and Del-R colonies suggests that physiological mechanisms of H. armigera resistance to Cry1Ac and deltamethrin do not confer cross-resistance to maysin.


Amputations and foot-related hospitalisations disproportionately affect dialysis patients.
Patients with diabetes have increased risk for foot ulcers, amputations and hospitalisations. We evaluated a closed cohort of patients with diabetes and established risk factors in two high risk groups: (i) dialysis patients and (ii) patients with previous foot ulceration. We used claims data for diabetes (ICD-9 250.X), ulceration (ICD-9 707·10, 707·14 and 707·15) and dialysis (CPT 90935-90937) from the Scott and White Health Plan to identify 150 consecutive patients with diabetes on dialysis (dialysis group) and 150 patients with a history of foot ulceration (ulcer history group). We verified these diagnoses by manually reviewing corresponding electronic medical records. Each patient was provided 30 months follow-up period. The incidence of foot ulcers was the same in dialysis patients and patients with an ulcer history (210 per 1000 person-years). The amputation incidence rate was higher in dialysis patients (58·0 versus 13·3, P < 0·001). Hospital admission was common in both study groups. The incidence of hospitalisation was higher in the ulcer history group (477·3 versus 381·3, P < 0·001); however, there were more foot-related hospital admissions in the dialysis group (32·9% versus 14·0%, P < 0·001) during the 30-month evaluation period. The incidence of ulcers, amputations and all-cause hospitalisations is high in persons with diabetes and a history of foot ulceration or on dialysis treatment; however, those on dialysis treatment have disproportionately higher rates of foot-related hospitalisations. Intervention strategies to reduce the burden of diabetic foot disease must target dialysis patients as a high-risk group.


[Value of ultrasonics in diagnosing the onset of lymphomatous liver lesions].
Ultrasound and anatomic findings were compared in 128 cases of lymphoma [87 non-Hodgkin lymphomas (NHL), 41 Hodgkin's disease (HD). Lymphomatous or other hepatic anomalies were observed in 40 cases (33 NHL, 7 HD). The 33 NHL lesions included 22 lymphomatous sites and 13 non-lymphomatous pathologies (in particular 5 biliary cysts and 4 cavernous liver hemangiomas). The 7 cases of hepatic anomalies in patients with HD included 6 specific disease sites and 1 cavernous hemangioma. For NHL, the sensitivity of ultrasonography in our series was 72.7% and the specificity 97%. In HD, the sensitivity was 66.6% and the specificity 100%. The value of ultrasound for the detection of lymphomatous liver lesions appears comparable to the results obtained with CT published in the literature; ultrasonography should thus be the first examination performed for subdiaphragmatic exploration during the investigation and follow-up of lymphomas.


A phase I study of foretinib plus erlotinib in patients with previously treated advanced non-small cell lung cancer: Canadian cancer trials group IND.196.
MET and AXL mediate resistance to EGFR TKI in NSCLC. Foretinib, a MET/RON/AXL/TIE-2/VEGFR kinase inhibitor may overcome EGFR kinase resistance. This dose escalation study combined foretinib and erlotinib in advanced pretreated NSCLC patients. The primary endpoint was to define the RP2D of foretinib plus erlotinib as continuous oral daily dosing. Secondary objectives included safety, pharmacokinetics, response and potential biomarkers of response including EGFR, KRAS genotype, MET, AXL expression, and circulating HGF levels. Erlotinib (E100-150 mg) was commenced on day 1 cycle 1; if well tolerated, foretinib (F30-45 mg) was added on day 15 cycle 1, using standard 3+3 dose escalation. Of 31 patients enrolled in 3 dose levels, 6 were inevaluable for DLT and replaced. DLT occurred in 3/15 patients at DL2 (E150 mg, F30 mg): Gr3 pain, mucositis, fatigue and rash. Cycle 1 DLT was not seen at DL3 (E150 mg, F45 mg) but 27% experienced dose reduction/interruption. Adverse events in ≥20% included diarrhea, fatigue, anorexia, dry skin, rash and hypertension. No PK interaction was seen with the combination. RP2D was defined as erlotinib 150 mg daily x 14 days with foretinib 30 mg added on day 15 (continuous dosing in 28-day cycles). Responses were seen in 17.8% of response evaluable patients (5/28). In 18 samples, baseline MET expression uncontrolled for EGFR genotype appeared associated with response. AXL expression was associated with neither EGFR mutation nor response. Combining foretinib and erlotinib demonstrated response in unselected advanced NSCLC but also incremental toxicity. Future development will require molecular patient selection.


Classification of climate-change-induced stresses on biological diversity.
Conservation actions need to account for and be adapted to address changes that will occur under global climate change. The identification of stresses on biological diversity (as defined in the Convention on Biological Diversity) is key in the process of adaptive conservation management. We considered any impact of climate change on biological diversity a stress because such an effect represents a change (negative or positive) in key ecological attributes of an ecosystem or parts of it. We applied a systemic approach and a hierarchical framework in a comprehensive classification of stresses to biological diversity that are caused directly by global climate change. Through analyses of 20 conservation sites in 7 countries and a review of the literature, we identified climate-change-induced stresses. We grouped the identified stresses according to 3 levels of biological diversity: stresses that affect individuals and populations, stresses that affect biological communities, and stresses that affect ecosystem structure and function. For each stress category, we differentiated 3 hierarchical levels of stress: stress class (thematic grouping with the coarsest resolution, 8); general stresses (thematic groups of specific stresses, 21); and specific stresses (most detailed definition of stresses, 90). We also compiled an overview of effects of climate change on ecosystem services using the categories of the Millennium Ecosystem Assessment and 2 additional categories. Our classification may be used to identify key climate-change-related stresses to biological diversity and may assist in the development of appropriate conservation strategies. The classification is in list format, but it accounts for relations among climate-change-induced stresses.


Association of immune activation with intravenous heroin use and methadone treatment in HIV-1 seropositive and seronegative subjects.
This study evaluated the effect of continued heroin use during methadone treatment on serum neopterin levels in 40 human immunodeficiency virus (HIV) type 1 seropositive (HIV+) and 70 seronegative (HIV-) intravenous drug users (IVDUs). Persistent drug use, determined by urinary evidence of opiates, was more common in HIV+ than in HIV- IVDUs (p = 0.01). Serum neopterin concentration, an indicator of increased probability of progression to AIDS, was elevated in HIV+ IVDUs, p < 0.0001 (mean 16.0 nmol/L) compared to that of HIV- IVDUs (mean 10.0 nmol/L) and in persistent IV drug users compared with abstainers, p < 0.0001. The effect of drug use and HIV status on serum neopterin was not explained by differences in methadone treatment, age, sex, or total years of i.v. drug use. Neopterin decreased in 25 IVDUs, regardless of HIV serostatus, treated with methadone for an average of 1.5 years.


Anterior-Segment Optical Coherence Tomography for Predicting Postoperative Outcomes After Trabeculectomy.
To determine whether anterior-segment optical coherence tomography (AS-OCT) can be used to predict post-trabeculectomy bleb outcomes. We divided 58 eyes of 47 trabeculectomy patients into success or failure groups based on their status at 12 months after surgery. We then compared various AS-OCT measurement parameters between the two groups at 1 and 2 weeks and 1, 3, 6, and 12 months. We also analyzed the early post-trabeculectomy bleb parameters with multiple logistic regression, stepwise multiple regression, and the receiver operating characteristic (ROC) curve, to evaluate the power of these parameters to predict long-term outcomes. Intraocular pressure 3 or more months after trabeculectomy was significantly lower in the success group than the failure group (all: P < 0.0016). Cleft volume was significantly higher 6 or more months after trabeculectomy in the success group than the failure group (P = 0.0027 and <0.0016). Reflectivity of the bleb wall was significantly higher in the failure group than the success group at 2 weeks and all later time points (all: P < 0.0016). Reflectivity of the bleb wall at 2 weeks after trabeculectomy was a risk factor for failure, with an odds ratio (OR) for failure of 2.48 (95% confidence interval, 1.31-4.68, increasing per 10 AU). The area under the ROC curve for reflectivity of the bleb wall at 2 weeks after trabeculectomy was 0.775 when the cutoff value was set at 122.8, with sensitivity, specificity, and OR of 78.3%, 80.0%, and 14.4, respectively. A stepwise multiple regression analysis showed that reflectivity of the bleb wall at 2 weeks was an independent factor indicating postoperative bleb survival period (β = -0.39, P = 0.007). Reflectivity of the bleb wall, measured by AS-OCT, may be an early post-trabeculectomy predictor of bleb outcome.


Implementation of the new EU IVD regulation - urgent initiatives are needed to avert impending crisis.
Laboratory medicine in the European Union is at the dawn of a regulatory revolution as it reaches the end of the transition from IVDD 98/79/EC (https://eur-lex.eur-opa.eu/legal-content/EN/TXT/?uri=CELEX%3A31998L0079&qid=1628781352814) to IVDR 2017/746 https://eur-lex.europa.eu/eli/reg/2017/746. Without amendments and contingency plans, implementation of the IVDR in May 2022 will lead the healthcare sector into uncharted waters due to unpreparedness of the EU regulatory infrastructure. Prospective risk analyses were not made by the European Commission, and if nothing happens it can be anticipated that the consequences will impact all stakeholders of the medical test pipeline, may seriously harm patients and may prevent caregivers from making appropriate clinical decisions due to non-availability of medical tests. Finally, it also may discourage manufacturers and academia from developing specialty tests, thereby hampering innovation in medical diagnostic care. We hereby inform laboratory professionals about the imminent diagnostic collapse using testimonies from representative stakeholders of the diagnostic supply chain and from academia developing innovative in-house tests in domains of unmet clinical needs. Steps taken by the EFLM Task Force on European Regulatory Affairs, under the umbrella of the Biomedical Alliance in Europe, will be highlighted, as well as the search for solutions through dialogue with the European Commission. Although we recognize that the IVDR promotes positive goals such as increased clinical evidence, surveillance, and transparency, we need to ensure that the capabilities of the diagnostic sector are not damaged by infrastructural unpreparedness, while at the same time being forced to submit to a growing bureaucratic and unsupportive structure that will not support its "droit d'exister".


Impaired fertility and spermiogenetic disorders with loss of cell adhesion in male mice expressing an interfering Rap1 mutant.
The guanosine trisphosphatase Rap1 serves as a critical player in signal transduction, somatic cell proliferation and differentiation, and cell-cell adhesion by acting through distinct mechanisms. During mouse spermiogenesis, Rap1 is activated and forms a signaling complex with its effector, the serine-threonine kinase B-Raf. To investigate the functional role of Rap1 in male germ cell differentiation, we generated transgenic mice expressing an inactive Rap1 mutant selectively in differentiating spermatids. This expression resulted in a derailment of spermiogenesis due to an anomalous release of immature round spermatids from the seminiferous epithelium within the tubule lumen and in low sperm counts. These spermiogenetic disorders correlated with impaired fertility, with the transgenic males being severely subfertile. Because mutant testis exhibited perturbations in ectoplasmic specializations (ESs), a Sertoli-germ cell-specific adherens junction, we searched for expression of vascular endothelial cadherin (VE-cadherin), an adhesion molecule regulated by Rap1, in spermatogenic cells of wild-type and mutant mice. We found that germ cells express VE-cadherin with a timing strictly related to apical ES formation and function; immature, VE-cadherin-positive spermatids were, however, prematurely released in the transgenic testis. In conclusion, interfering with Rap1 function during spermiogenesis leads to reduced fertility by impairment of germ-Sertoli cell contacts; our transgenic mouse provides an in vivo model to study the regulation of ES dynamics.


Insights into the catalytic mechanism of tyrosine phenol-lyase from X-ray structures of quinonoid intermediates.
Amino acid transformations catalyzed by a number of pyridoxal 5'-phosphate (PLP)-dependent enzymes involve abstraction of the Calpha proton from an external aldimine formed between a substrate and the cofactor leading to the formation of a quinonoid intermediate. Despite the key role played by the quinonoid intermediates in the catalysis by PLP-dependent enzymes, limited accurate information is available about their structures. We trapped the quinonoid intermediates of Citrobacter freundii tyrosine phenol-lyase with L-alanine and L-methionine in the crystalline state and determined their structures at 1.9- and 1.95-A resolution, respectively, by cryo-crystallography. The data reveal a network of protein-PLP-substrate interactions that stabilize the planar geometry of the quinonoid intermediate. In both structures the protein subunits are found in two conformations, open and closed, uncovering the mechanism by which binding of the substrate and restructuring of the active site during its closure protect the quinonoid intermediate from the solvent and bring catalytically important residues into positions suitable for the abstraction of phenol during the beta-elimination of L-tyrosine. In addition, the structural data indicate a mechanism for alanine racemization involving two bases, Lys-257 and a water molecule. These two bases are connected by a hydrogen bonding system allowing internal transfer of the Calpha proton.


Development of the T-ALLiPSC-based therapeutic cancer vaccines for T-cell acute lymphoblastic leukemia.
The T-cell acute lymphoblastic leukemia (T-ALL) is a kind of hematological malignancy in children. Despite the significant improvement in the cure rate of T-ALL upon treatment with chemotherapy regimens, steroids, and allotransplantation there are relapses. This study focuses on the tumor-specific therapeutic vaccines derived from the induced pluripotent stem cells (iPSC) to address the issue of T-ALL recurrence. Patient-derived tumor cells and healthy donor cells were reprogrammed into the iPSCs and the RNA-seq data of the T-ALL-iPSCs and H-iPSCs were analyzed. In vitro, the whole-cell lysate antigens of iPSCs were prepared to induce the dendritic cells (DC) maturation, which in turn stimulated the tumor-specific T cells to kill the T-ALL tumor cells (Jurkat, CCRF-CEM, MOLT-4). The cytotoxic T lymphocyte stimulated by the DC-loaded T-ALL-iPSC-derived antigens showed specific cytotoxicity against the T-ALL cells in vitro. In conclusion, the T-ALL-iPSC-based therapeutic cancer vaccine can elicit a specific anti-tumor effect on T-ALL.


Contraction inhibits insulin-stimulated insulin receptor substrate-1/2-associated phosphoinositide 3-kinase activity, but not protein kinase B activation or glucose uptake, in rat muscle.
The initial stages of insulin-stimulated glucose uptake are thought to involve tyrosine phosphorylation of insulin receptor substrates (IRSs), which recruit and activate phosphoinositide 3-kinase (PI 3-kinase), leading to the activation of protein kinase B (PKB) and other downstream effectors. In contrast, contraction stimulates glucose uptake via a PI 3-kinase-independent mechanism. The combined effects of insulin and contraction on glucose uptake are additive. However, it has been reported that contraction causes a decrease in insulin-stimulated IRS-1-associated PI 3-kinase activity. To investigate this paradox, we have examined the effects of contraction on insulin-stimulated glucose uptake and proximal insulin-signalling events in isolated rat epitrochlearis muscle. Stimulation by insulin or contraction produced a 3-fold increase in glucose uptake, with the effects of simultaneous treatment by insulin and contraction being additive. Wortmannin completely blocked the additive effect of insulin in contracting skeletal muscle, indicating that this is a PI 3-kinase-dependent effect. Insulin-stimulated recruitment of PI 3-kinase to IRS-1 was unaffected by contraction; however, insulin produced no discernible increase in PI 3-kinase activity in IRS-1 or IRS-2 immunocomplexes in contracting skeletal muscle. Consistent with this, contraction inhibited insulin-stimulated p70(S6K) activation. In contrast, insulin-stimulated activation of PKB was unaffected by contraction. Thus, in contracting skeletal muscle, insulin stimulates glucose uptake and activates PKB, but not p70(S6K), by a PI 3-kinase-dependent mechanism that is independent of changes in IRS-1- and IRS-2-associated PI 3-kinase activity.


Functional disturbance of marrow stromal microenvironment in the myelodysplastic syndromes.
The potential contribution of abnormal marrow stromal function to ineffective haemopoiesis in the myelodysplastic syndromes is unclear. We have compared the ability of stromal layers from normal (n = 7) and myelodysplastic (n = 9) marrow to alter proliferation and survival of the granulocyte-macrophage colony-stimulating factor/interleukin-3-dependent cell line F-36P. Co-cultures for 72 h in the absence of exogenous cytokines were either in direct contact with stroma or separated by transwell inserts. On normal stromal layers, the ratio of adherent F-36P cells relative to stromal cells increased from a mean of 0.2 +/- 0.01 (s.d.) at 4 h of co-culture to 0.34 +/- 0.08 after 72 h (n = 7). Corresponding values on myelodysplastic stroma (0.2 +/- 0.02 at 4 h and 0.35 +/- 0.05 at 72 h; n = 9) indicated that the ability of myelodysplastic stromal layers to regulate short-term proliferation of F-36P cells may be similar to normal. Apoptosis of F-36P cells was quantified after co-culture with normal or myelodysplastic stroma: results from myelodysplastic co-cultures were standardized as a fraction of values from co-cultures with paired normal stroma (apoptotic ratio). Augmented apoptosis of F-36P cells was detected in 8/9 co-cultures with myelodysplastic stroma (mean = 15.7 +/- 9.7%, n = 9), compared with corresponding normal stroma (mean = 12.4 +/- 4.6%, n = 7, P < 0.05) with a mean apoptotic ratio of 1.4 +/- 0.5 (P < 0.05). There was no correlation between stroma-related apoptosis and FAB type, tumour necrosis factor-alpha concentrations in the culture supernatant or numbers of stromal macrophages, and no evidence of involvement of the Fas pathway. Increased apoptosis was detected in cells grown in transwell inserts over stroma (23.8 +/- 3%, n = 5) compared to adherent cells in cultures with normal stromal layers, but this survival difference was not observed in co-cultures with myelodysplastic stroma. These results suggest that abnormal stromal function in patients with myelodysplastic syndromes may contribute to increased apoptosis of haemopoietic cells within the marrow microenvironment. The effect appears to be dependent on close cellular contact, rather than the release of soluble factors, but the exact mechanism remains unclear.


Evaluation of the combination of nelarabine and fludarabine in leukemias: clinical response, pharmacokinetics, and pharmacodynamics in leukemia cells.
A pilot protocol was designed to evaluate the efficacy of fludarabine with nelarabine (the prodrug of arabinosylguanine [ara-G]) in patients with hematologic malignancies. The cellular pharmacokinetics was investigated to seek a relationship between response and accumulation of ara-G triphosphate (ara-GTP) in circulating leukemia cells and to evaluate biochemical modulation of cellular ara-GTP metabolism by fludarabine triphosphate. Nine of the 13 total patients had indolent leukemias, including six whose disease failed prior fludarabine therapy. Two patients had T-acute lymphoblastic leukemia, one had chronic myelogenous leukemia, and one had mycosis fungoides. Nelarabine (1.2 g/m(2)) was infused on days 1, 3, and 5. On days 3 and 5, fludarabine (30 mg/m(2)) was administered 4 hours before the nelarabine infusion. Plasma and cellular pharmacokinetic measurements were conducted during the first 5 days. Seven patients had a partial or complete response, six of whom had indolent leukemias. The disease in four responders had failed prior fludarabine therapy. The median peak intracellular concentrations of ara-GTP were significantly different (P =.001) in responders (890 micromol/L, n = 6) and nonresponders (30 micromol/L, n = 6). Also, there was a direct relationship between the peak fludarabine triphosphate and ara-GTP in each patient (r = 0.85). The cellular elimination of ara-GTP was slow (median, 35 hours; range, 18 to > 48 hours). The ratio of ara-GTP to its normal counterpart, deoxyguanosine triphosphate, was higher in each patient (median, 42; range, 14 to 1,092) than that of fludarabine triphosphate to its normal counterpart, deoxyadenosine triphosphate (median, 2.2; range, 0.2 to 27). Fludarabine plus nelarabine is an effective, well-tolerated regimen against leukemias. Clinical responses suggest the need for further exploration of nelarabine against fludarabine-refractory diseases. Determination of ara-GTP levels in the target tumor population may provide a prognostic test for the activity of nelarabine.


Prevalence of Adverse Childhood Experiences From the 2011-2014 Behavioral Risk Factor Surveillance System in 23 States.
Early adversity is associated with leading causes of adult morbidity and mortality and effects on life opportunities. To provide an updated prevalence estimate of adverse childhood experiences (ACEs) in the United States using a large, diverse, and representative sample of adults in 23 states. Data were collected through the Behavioral Risk Factor Surveillance System (BRFSS), an annual, nationally representative telephone survey on health-related behaviors, health conditions, and use of preventive services, from January 1, 2011, through December 31, 2014. Twenty-three states included the ACE assessment in their BRFSS. Respondents included 248 934 noninstitutionalized adults older than 18 years. Data were analyzed from March 15 to April 25, 2017. The ACE module consists of 11 questions collapsed into the following 8 categories: physical abuse, emotional abuse, sexual abuse, household mental illness, household substance use, household domestic violence, incarcerated household member, and parental separation or divorce. Lifetime ACE prevalence estimates within each subdomain were calculated (range, 1.00-8.00, with higher scores indicating greater exposure) and stratified by sex, age group, race/ethnicity, annual household income, educational attainment, employment status, sexual orientation, and geographic region. Of the 214 157 respondents included in the sample (51.51% female), 61.55% had at least 1 and 24.64% reported 3 or more ACEs. Significantly higher ACE exposures were reported by participants who identified as black (mean score, 1.69; 95% CI, 1.62-1.76), Hispanic (mean score, 1.80; 95% CI, 1.70-1.91), or multiracial (mean score, 2.52; 95% CI, 2.36-2.67), those with less than a high school education (mean score, 1.97; 95% CI, 1.88-2.05), those with income of less than $15 000 per year (mean score, 2.16; 95% CI, 2.09-2.23), those who were unemployed (mean score, 2.30; 95% CI, 2.21-2.38) or unable to work (mean score, 2.33; 95% CI, 2.25-2.42), and those identifying as gay/lesbian (mean score 2.19; 95% CI, 1.95-2.43) or bisexual (mean score, 3.14; 95% CI, 2.82-3.46) compared with those identifying as white, those completing high school or more education, those in all other income brackets, those who were employed, and those identifying as straight, respectively. Emotional abuse was the most prevalent ACE (34.42%; 95% CI, 33.81%-35.03%), followed by parental separation or divorce (27.63%; 95% CI, 27.02%-28.24%) and household substance abuse (27.56%; 95% CI, 27.00%-28.14%). This report demonstrates the burden of ACEs among the US adult population using the largest and most diverse sample to date. These findings highlight that childhood adversity is common across sociodemographic characteristics, but some individuals are at higher risk of experiencing ACEs than others. Although identifying and treating ACE exposure is important, prioritizing primary prevention of ACEs is critical to improve health and life outcomes throughout the lifespan and across generations.


New therapeutic approach in inflammatory bowel disease.
In the last decade, biologic agents, in particular infliximab and adalimum-ab, have deeply changed the therapeutic armamentarium of inflammatory bowel disease (IBD). However, these drugs have a number of contraindications and side-effects that physicians should know so to avoid and eventually manage them. Another important issue is the early introduction of immunomodulators and biologics in the therapeutic algorithm of IBD, the so called "top-down" approach compared to the traditional "step-up" approach. In this review, the indications to the use of anti-TNF-alpha molecules in IBD are briefly reported and the potential benefits and disadvantages of a more aggressive approach are discussed.


Mercury exposure and child development outcomes.
Mercury is ubiquitous in the global environment, ensuring universal exposure. Some forms of mercury are especially neurotoxic, including clinical signs at high doses. However, typical human exposures occur at low to moderate doses. Only limited data about neurotoxicity at low doses are available, and scientists differ in their interpretation. Dose-response data on neurodevelopment are particularly limited. Despite or perhaps because of the lack of sufficient or consistent scientific data, public concern about a link between mercury exposure and developmental disabilities has been rising. After reviewing the data, the US Environmental Protection Agency proposed a reference dose (an estimate of a daily dose that is likely to be without a risk of adverse effects over a lifetime) for methyl mercury that is substantially lower than previous guidelines from the World Health Organization, the US Agency for Toxic Substances and Disease Registry, and the US Food and Drug Administration. Some questions have been raised about the Environmental Protection Agency's guidelines, but the issue remains unresolved. Meanwhile, consumer groups have raised questions about the potential link between mercury exposure and autism spectrum disorders as well as other adverse neurodevelopmental outcomes. This hypothesis has prompted some parents to seek regulatory, legal, or medical remedies in the absence of firm evidence. This article reviews what is known about mercury neurotoxicity and neurodevelopmental risk. Our intent is to focus the debate about mercury on 1) additional research that should be sought and 2) defining the principal issues that public policy makers face.


Foretinib Inhibits Cancer Stemness and Gastric Cancer Cell Proliferation by Decreasing CD44 and c-MET Signaling.
CD44 isoforms are highly expressed in cancer stem cells, initiating tumor growth and sustaining tumor self-renewal. Among these isoforms, CD44 variant 9 (CD44v9) is overexpressed in chronic inflammation-induced cancer. CD44 and the mesenchymal-to-epithelial transition (MET) receptor tyrosine kinase are coactivated in some gastric cancers (GCs). In this study, we characterized MET and CD44 expression and signaling in human GC cell lines and analyzed differences in the susceptibility of these lines to foretinib. We analyzed cell viability and the rate of apoptotic cells using MTS assays and flow cytometry, respectively. Gene and protein expression were assessed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and immunoblotting, respectively. Foretinib treatment resulted in dose-dependent inhibition of growth in c-MET-amplified MKN45 and SNU620 cells with concomitant induction of apoptosis, but not in c-MET-reduced MKN28 and AGS cells. Foretinib treatment also significantly reduced phosphor-c-MET, phosphor-AKT, beta-catenin, and COX-2 protein expression in MKN45 and SNU620 cells. Interestingly, foretinib significantly reduced CD44, CD44v9, COX-2, OCT3/4, CCND1, c-MYC, VEGFA, and HIF-1a gene expression in CD44 and MET coactivated MKN45 cells and increased CD44s gene expression; in contrast, these drugs were only slightly active against SNU620 cells. The results of this study indicate that foretinib could be a therapeutic agent for the prevention or treatment of GCs positive for CD44v9 and c-MET.


Substrate turnover and inhibitor binding as selection parameters in directed evolution of blood coagulation factor Xa.
A library of blood coagulation factor Xa (FXa)-trypsin hybrid proteases was generated and displayed on phage for selection of derivatives with the domain "architecture" of trypsin and the specificity of FXa. Selection based on binding to soybean trypsin inhibitor only provided enzymatically inactive derivatives, due to a specific mutation of serine 195 of the catalytic triad to a glycine, revealing a significant selection pressure for proteolytic inactive derivatives. By including a FXa peptide substrate in the selection mixture, the majority of the clones had retained serine at position 195 and were enzymatically active after selection. Further, with the inclusion of bovine pancreatic trypsin inhibitor, in addition to the peptide substrate, the selected clones also retained FXa specificity after selection. This demonstrates that affinity selection combined with appropriate deselection provides a simple strategy for selection of enzyme derivatives that catalyse a specific reaction.


Glyphosate, AMPA and glyphosate-based herbicide exposure leads to GFAP, PCNA and caspase-3 increased immunoreactive area on male offspring rat hypothalamus.
Glyphosate, aminomethylphosphonic acid (AMPA), and glyphosate-based herbicides altered the neuroendocrine axis, the content of brain neurotransmitters, and behavior in experimental animal models. Glyphosate alone, AMPA or Roundup® Active were administered to postpartum female rats, from P0 to P10, and their water consumption was measured daily. The immunoreactivity for glial fibrillary acidic protein (GFAP), proliferating cell nuclear antigen (PCNA) and caspase-3 was measured in the anterior, medial preoptic, periventricular, supraoptic and lateroanterior hypothalamic nuclei of P0-P10 male pups after exposure, via lactation, to these xenobiotics. Puppies exposed to glyphosate had a moderate level of GFAP with no overlapping astrocyte processes, but this overlapping was observed after Roundup® Active or AMPA exposure. After being exposed to Roundup® Active or AMPA, PCNA-positive cells with strong immunoreactivity were found in some hypothalamic nuclei. Cells containing caspase-3 were found in all hypothalamic nuclei studied, but the labeling was stronger after Roundup® Active or AMPA exposure. Xenobiotics significantly increased the immunoreactivity area for all of the markers studied in the majority of cases (p<0.05). AMPA or Roundup® Active treated animals had a greater area of PCNA immunoreactivity than control or glyphosate alone treated animals (p<0.05). The effects observed after xenobiotic exposure were not due to increased water intake. The increased immunoreactivity areas observed for the markers studied suggest that xenobiotics induced a neuro-inflammatory response, implying increased cell proliferation, glial activation, and induction of apoptotic pathways. The findings also show that glyphosate metabolites/adjuvants and/or surfactants present in glyphosate commercial formulations had a greater effect than glyphosate alone. In summary, glyphosate, AMPA, and glyphosate-based herbicides altered GFAP, caspase-3, and PCNA expression in the rat hypothalamus, altering the neuroendocrine axis.


Salt links dominate affinity of antibody HyHEL-5 for lysozyme through enthalpic contributions.
The binding of murine monoclonal antibody HyHEL-5 to lysozyme has been the subject of extensive crystallographic, computational, and experimental investigations. The complex of HyHEL-5 with hen egg lysozyme (HEL) features salt bridges between Fab heavy chain residue Glu(50), and Arg(45) and Arg(68) of HEL. This interaction has been predicted to play a dominant role in the association on the basis of molecular electrostatics calculations. The association of aspartic acid and glutamine mutants at position 50(H) of the cloned HyHEL-5 Fab with HEL and bobwhite quail lysozyme (BQL), an avian variant bearing an Arg(68) --> Lys substitution in the epitope, was characterized by isothermal titration calorimetry and sedimentation equilibrium. Affinities for HEL were reduced by 400-fold (E50(H)D) and 40,000-fold (E50(H)Q) (DeltaDeltaG degrees estimated at 4.0 and 6.4 kcal mol(-1), respectively). The same mutations reduce affinity for BQL by only 7- and 55-fold, respectively, indicating a reduced specificity for HEL. The loss of affinity upon mutation is in each case primarily due to an unfavorable change in the enthalpy of the interaction; the entropic contribution is virtually unchanged. An enthalpy-entropy compensation exists for each interaction; DeltaH degrees decreases, while DeltaS degrees increases with temperature. The DeltaCp for each mutant interaction is less negative than the wild-type. Mutant-cycle analysis suggests the mutations present in the HyHEL-5 Fab mutants are linked to those present in the BQL with coupling energies between 3 and 4 kcal mol(-1).


Relative opioid efficacy is determined by the complements of the G protein-coupled receptor desensitization machinery.
G protein-coupled receptor regulation by G protein-coupled receptor kinases and beta-arrestins can lead to desensitization and subsequent internalization of the receptor. In in vitro and cellular systems, beta-arrestins do not seem to play a major role in regulating micro opioid receptor (microOR) responsiveness. Removal of the betaarrestin2 (betaarr2) gene in mice leads paradoxically to enhanced and prolonged microOR-mediated antinociception. The betaarr2 knockout (betaarr2-KO) mice also fail to develop morphine antinociceptive tolerance in the hot-plate test, further indicating that the betaarr2 protein plays an essential role in microOR regulation in vivo. In this study, the contribution of betaarr2 to the regulation of the microOR was examined in both human embryonic kidney 293 cells and in betaarr2-KO mice after treatment with several opiate agonists. A green fluorescent protein tagged betaarr2 was used to assess receptor-betaarr2 interactions in living cells. Opiate agonists that induced robust betaarr2-green fluorescent protein translocation produced similar analgesia profiles in wild-type and betaarr2-KO mice, whereas those that do not promote robust betaarr2 recruitment, such as morphine and heroin, produce enhanced analgesia in vivo. In this report, we present a rationale to explain the seemingly paradoxical relationship between beta-arrestins and microOR regulation wherein morphine-like agonists fail to promote efficient internalization and resensitization of the receptor.


Passive smoke exposure and circulating carotenoids in the CARDIA study.
Our objective was to assess associations between passive smoke exposure in various venues and serum carotenoid concentrations. CARDIA is an ongoing longitudinal study of the risk factors for subclinical and clinical cardiovascular disease. At baseline in 1985/1986, serum carotenoids were assayed and passive smoke exposure inside and outside of the home and diet were assessed by self-report. Our analytic sample consisted of 2,633 black and white non-smoking adults aged 18-30 years. Greater total passive smoke exposure was associated with lower levels of the sum of the three provitamin A carotenoids, alpha-carotene, beta-carotene, and beta-cryptoxanthin (-0.048 nmol/l per hour of passive smoke exposure, p = 0.001), unassociated with lutein/zeaxanthin, and associated with higher levels of lycopene (0.027 nmol/l per hour of passive smoke exposure, p = 0.010) after adjustment for demographics, diet, lipid profile, and supplement use. Exposure in both home and non-home spaces was also associated with lower levels of the provitamin A carotenoid index. Cross-sectionally, in 1985/86, passive smoke exposure in various venues was associated with reduced levels of provitamin A serum carotenoids.