Antibodies against Noncatalytic B Subunit of Factor XIII Inhibit Activation of Factor XIII and Fibrin Crosslinking.
Coagulation factor XIII (FXIII) is a proenzyme of plasma transglutaminase. It comprises two catalytic A subunits (FXIII-A) and two carrier B subunits (FXIII-B). We previously reported that alloantibodies against FXIII-B could promote FXIII clearance in a patient with congenital FXIII-B deficiency who had received infusions of plasma-derived human FXIII (A2B2 heterotetramer). We aimed to investigate whether anti-FXIII-B antibodies affect the catalytic function of FXIII. FXIII activation and fibrin crosslinking were examined in the presence of patient plasma, isolated patient IgG, or rat anti-FXIII-B monoclonal antibodies. Alloantibody levels were increased by repeated infusions of plasma-derived A2B2 heterotetramer, which enhanced binding to the functionally important FXIII-B sushi domains. The patient plasma strongly inhibited cleavage of the FXIII-A activation peptide, amine incorporation, and fibrin crosslinking in normal plasma. Furthermore, anti-FXIII-B alloantibodies blocked the formation of the complex of FXIII-B with FXIII-A, and fibrinogen. Rat monoclonal antibodies against the 10th sushi domain of FXIII-B inhibited the incorporation of FXIII-B to fibrin, FXIII activation (i.e., cleavage of FXIII-A activation peptide), and ultimately fibrin crosslinking in normal plasma, independent of their effect on heterotetramer assembly with FXIII-A. Alloantibody binding to the A2B2 heterotetramer blocked the access of thrombin to the FXIII-A cleavage site, as indicated by the reaction of the alloantibodies to the A2B2 heterotetramer and FXIII-B, but not to FXIII-A. Anti-FXIII-B antibodies binding to the A2B2 heterotetramer and FXIII-B inhibited FXIII activation and its crosslinking function despite being directed against its noncatalytic subunit (FXIII-B).


Genetic stability of live attenuated vaccines against potentially pandemic influenza viruses.
Ensuring genetic stability is a prerequisite for live attenuated influenza vaccine (LAIV). This study describes the results of virus shedding and clinical isolates' testing of Phase I clinical trials of Russian LAIVs against potentially pandemic influenza viruses in healthy adults. Three live attenuated vaccines against potentially pandemic influenza viruses, H2N2 LAIV, H5N2 LAIV and H7N3 LAIV, generated by classical reassortment in eggs, were studied. For each vaccine tested, subjects were randomly distributed into two groups to receive two doses of either LAIV or placebo at a 3:1 vaccine/placebo ratio. Nasal swabs were examined for vaccine virus shedding by culturing in eggs and by PCR. Vaccine isolates were tested for temperature sensitivity and cold-adaptation (ts/ca phenotypes) and for nucleotide sequence. The majority of nasal wash positive specimens were detected on the first day following vaccination. PCR method demonstrated higher sensitivity than routine virus isolation in eggs. None of the placebo recipients had detectable vaccine virus replication. All viruses isolated from the immunized subjects retained the ts/ca phenotypic characteristics of the master donor virus (MDV) and were shown to preserve all attenuating mutations described for the MDV. These data suggest high level of vaccine virus genetic stability after replication in humans. During manufacture process, no additional mutations occurred in the genome of H2N2 LAIV. In contrast, one amino acid change in the HA of H7N3 LAIV and two additional mutations in the HA of H5N2 LAIV manufactured vaccine lot were detected, however, they did not affect their ts/ca phenotypes. Our clinical trials revealed phenotypic and genetic stability of the LAIV viruses recovered from the immunized volunteers. In addition, no vaccine virus was detected in the placebo groups indicating the lack of person-to-person transmission. LAIV TRIAL REGISTRATION at ClinicalTrials.gov: H7N3-NCT01511419; H5N2-NCT01719783; H2N2-NCT01982331.


Survey of opioid use in chronic nonmalignant pain patients.
Opioids have been accepted as appropriate treatment for acute and cancer pain, but remain controversial for use with chronic nonmalignant pain. Clinicians are concerned about efficacy, tolerance, addiction, and unwanted side effects. The aim of this study was to survey chronic pain patients who were taking opioids for their pain, to determine the incidence of these adverse conditions. Two hundred seventeen patients who were being treated for their pain at two different pain centers completed a medication questionnaire. The most common diagnosis was low back pain. One hundred twelve patients reported taking oral opioids for their pain. Of the patients who reported taking opioids for their chronic pain, 83% felt that the opioids were moderately beneficial in relieving their pain; 25% felt that the opioid had not lost its ability to relieve the pain over time; 35% reported that they did not need to increase their medication; 36% expressed no fear of addiction or dependence; and 56% reported having no unwanted side effects. The results suggest that chronic nonmalignant pain patients taking opioids for their pain reported some tolerance, fear of addiction, and side effects when taking opioids. However, despite these concerns, some of these patients felt that opioid therapy was very beneficial. Further investigations are needed to determine which patient characteristics predict benefit from opioid therapy for nonmalignant pain.


How to Rethink the Fourteen-Day Rule.
Recently, attention has been drawn to the basic principles governing the use of human embryos in research: specifically, the so-called fourteen-day rule. This rule stipulates that human embryos should not be allowed to grow in vitro past fourteen days of development. For years, the fourteen-day limit was largely theoretical, since culture techniques were not sufficient to maintain embryos up to this point. Yet in the past year, research has suggested that growing embryos beyond fourteen days might be feasible and scientifically valuable. At the same time, work with pluripotent stem cells, including human PSCs, has shown that under certain conditions, they can form structures that recapitulate developmental features of the postimplantation embryo. This raises the possibility that PSCs could generate embryo-like structures in vitro, even "synthetic embryos," that might provoke moral concern but would not fall under most current embryo research policies. In countries that permit embryo research, the fourteen-day rule has long been the linchpin of an effective policy compromise between what remain deeply divided moral positions on the human embryo's status. It has also, particularly in the United Kingdom, been influential in establishing a bioethics public-policy process. Any moves to change the rule must consider not just the implications for the use of embryos but also the potential impact of this model of bioethical governance of science.


Proximity Labeling of Interacting Proteins: Application of BioID as a Discovery Tool.
Proteins perform biochemical functions by forming complexes, or protein-protein interactions (PPIs). Many different approaches such as phage display, yeast hybridization, etc. were developed to illustrate the PPIs, and disclose the composition and organization of protein complexes. However, none of these approaches are based on the real-time and in vivo PPI analysis. Proximity-dependent labeling (PDL) of interacting proteins has recently been proposed by taking advantage of several enzymes, which are capable of attaching the known reactive groups to the nearby proteins covalently. Among the PDL methods, BioID is the earliest and the most widely used one and has been upgraded from its prototype, making it an extremely convenient research tool. In this review, we describe the BioID technology development, its potential applications according to the nature of the target protein, and some recent efforts to circumvent the technical limitations. Moreover, some comparable PDL methods are introduced, including selective proteomic proximity labeling assay using tyramide, enzyme-mediated activation of radical sources, Proximity Labeling with Ascorbate Peroxidase, in vivo proximal labeling, etc., and we propose that systematic comparison of the working radius of these methods may be helpful to develop a tool box, from which the right method can be selected for a given target protein for PPI research.


Mesenchymal Stem Cells Modulate Differentiation of Myeloid Progenitor Cells During Inflammation.
Mesenchymal stem cells (MSCs) possess distinct immunomodulatory properties and have tremendous potential for use in therapeutic applications in various inflammatory diseases. MSCs have been shown to regulate pathogenic functions of mature myeloid inflammatory cells, such as macrophages and neutrophils. Intriguingly, the capacity of MSCs to modulate differentiation of myeloid progenitors (MPs) to mature inflammatory cells remains unknown to date. Here, we report the novel finding that MSCs inhibit the expression of differentiation markers on MPs under inflammatory conditions. We demonstrate that the inhibitory effect of MSCs is dependent on direct cell-cell contact and that this intercellular contact is mediated through interaction of CD200 expressed by MSCs and CD200R1 expressed by MPs. Furthermore, using an injury model of sterile inflammation, we show that MSCs promote MP frequencies and suppress infiltration of inflammatory cells in the inflamed tissue. We also find that downregulation of CD200 in MSCs correlates with abrogation of their immunoregulatory function. Collectively, our study provides unequivocal evidence that MSCs inhibit differentiation of MPs in the inflammatory environment via CD200-CD200R1 interaction. Stem Cells 2017;35:1532-1541.


Production of thiophenes from callus cultures of Tagetes patula L. and its mosquito larvicidal activity.
Calli were initiated from leaf tissues of T. patula, on Murashige and Skoog's medium supplemented with 2-4 dichlorophenoxacetic acid (2,4-D) and kinetin (Kn). The maximum accumulation of biomass was recorded on 6th week at 1.11 g. dry wt/culture. Maximum thiophene content was recorded on 4th week at 0.008% on dry wt basis in callus cultures of T. patula subjected to nutrient stress. Nitrogen stress induced 3-fold increase in thiophene production level in six weeks (0.024% on dry weight basis). The best hormonal supplementation required for thiophenes production was found to be 2,4-D (2.0 mg L(-1)) and kinetin (2.0 mg L(-1)). The thiophenes produced in callus cultures of T. patula showed larvicidal effect against mosquito larvae.


[Ring melanoma revealed by spontaneous hyphema].
Ring melanoma is a rare form of uveal melanoma characterized by the circumferential involvement of the ciliary body. Unilateral chronic and refractory glaucoma is a classic circumstance of diagnosis. We report a case of ring melanoma revealed by acute intraocular hypertonia secondary to spontaneous hyphema. Iris and iridocorneal angle were diffusely invaded by the tumor. The fundus was masked but B-scan ultrasonography revealed a voluminous ciliochoroidal tumor. The patient had been enucleated. Pathological examination confirmed the diagnosis of ring melanoma. The tumor involved angle structures. The patient died 1 year later because of metastasis. Acute or chronic ocular hypertonia is a classic but rare circumstance of uveal melanoma diagnosis. Many mechanisms exist: neovascular glaucoma, secondary angle closure, involvement of angle structures, and trabecular obstruction by tumor cells or pigment. Acute intraocular hypertonia secondary to hyphema is more exceptional. Our observation highlights that apart from the classic situation of acute angle closure glaucoma, intraocular hypertonia requires meticulous fundus examination, if necessary using B-scan ultrasonography.


Seasonality and geographical distribution of Kawasaki disease among Black children in the Southeast United States.
Kawasaki Disease (KD) is a leading cause of pediatric acquired heart disease in the United States, affecting up to 7,000 children annually. Seasonal variation, an epidemiological characteristic of KD, has previously been reported predominantly among Asian children; however, little is known about the epidemiology and seasonality of KD of Black children within the U.S. Electronic medical records were abstracted from 529 hospitalized KD patients admitted to a single tertiary center in Alabama between 2005 and 2019. Medical charts were reviewed to confirm KD diagnosis following American Heart Association criteria. Cases were stratified by the month of diagnosis date to assess seasonality, and statewide distribution of incidence is reported at county level using geographical spatial analysis. Comparisons were performed between Black patients and White patients with KD. The average number of KD cases per year was 35. Approximately, 60% were males and 44% were White children (N = 234), 45% were Black children (N = 240) and 11% were other races (N = 55). Black children were younger than White children at KD admission (median age 32 vs. 41 months respectively, p = 0.02). Overall, the highest rates of cases occurred between January and April. When stratifying by race, cases started to rise in December among White children with the highest rates between February and April with a peak in March. Among Black children cases were high during the winter season (January-April) with a peak in April. Similarly high rates also occurred in June, July and November. There were no differences in geographical distribution of cases by race. KD incidence among White children in Alabama follows a seasonal cycle similar to other regions in the U.S. However, sustained incidence and additional peaks outside of the usual KD seasonality were seen among Black children with KD. Further studies are needed to investigate differential triggers between races.


Time correlation function and finite field approaches to the calculation of the fifth order Raman response in liquid xenon.
The fifth order, two-dimensional Raman response in liquid xenon is calculated via a time correlation function (TCF) theory and the numerically exact finite field method. Both employ classical molecular dynamics simulations. The results are shown to be in excellent agreement, suggesting the efficacy of the TCF approach, in which the response function is written approximately in terms of a single classical multitime TCF.


Movement-related cortical potentials in ALS increase at lower and decrease at higher upper motor neuron burden scores.
Our aim was to investigate changes in movement-related cortical potentials (MRCPs) in ALS patients with different degrees of upper motor neuron (UMN) involvement. Since respiratory failure is the main cause of death in ALS, changes in inspiratory-related (sniffing) potentials were studied in addition to finger-flexion-related potentials. Subjects (21 ALS, 19 controls) performed two self-paced motor tasks while their EEGs were recorded. The first task required flexions of the right index finger and the second, brisk nasal inspirations. The early (BP1), late (BP2) and motor potential (MP) components of MRCPs were evaluated. Results showed that patients generated higher MRCPs than controls. However, this effect was most significant in the subgroup of patients with low UMN burden (LUB). The high UMN burden (HUB) subgroup did not differ from controls, but had significantly lower MP amplitudes than the LUB subgroup. Progressive UMN deterioration was associated with an initial increase, followed by a later decrease, in MP amplitudes in ALS. In conclusion, the increased MRCPs in LUB compared to HUB patients indicate different processes of ALS pathophysiology that force opposing changes in MRCP amplitudes.


The human vomeronasal system.
We studied the functional characteristics of the vomeronasal system in clinically normal adult subjects of both sexes (ages 20-45). Chemosensory substances were administered in punctate pulses in a continuous air stream from the tip of a multifunctional miniprobe, which contained a nonpolarizable electrode. Negative potentials with the characteristics of receptor potentials were recorded from the surface of the vomeronasal organ (VNO) and olfactory epithelium (OE) in response to certain substances defined here as vomeropherins (see definition in the introduction of the main text) and to olfactants. Stimulation of the VNO with femtomole amounts of vomeropherins produced a local depolarization with the characteristics of a receptor potential. The same substances produced only a small response from the OE, and no response from the nasal respiratory mucosa. Three vomeropherins PH15, PH78, and PH84 were particularly well recognized by the VNO of most male subjects (p < .01; n = 30). Substances PH30, PH56, and PH94B, produced similar effects in the VNO of most female subjects (p < .01; n = 30). Responses to virtually all vomeropherins exhibited a sexual dimorphism. Stimulation of the OE with the same quantity of odorants 1,8-cineole and l-carvone produced depolarization of 6.8 +/- 2.6 mV, but little or no response in the VNO. Therefore, the human VNO seems to have a unique specificity for certain chemosensory substances when compared to the OE. Administration of PH15 and PH78 to the VNO of male subjects (but not to female subjects) significantly increased electrodermal activity (p < .02) and skin temperature (p < .01). On the other hand, administration of PH84 to the VNO of male subjects decreased skin temperature but had little effect on electrodermal activity. Autonomic changes were accompanied by an increased percentage of alpha-cortical activity for all three vomeropherins. In female subjects (but not in male subjects) vomeropherins PH56 and PH94B significantly increased electrodermal activity (p < .01), skin temperature (p < .01), and alpha-cortical activity (p < .01). Local application of the olfactants 1,8-cineole and l-carvone to the VNO did not trigger autonomic responses or significant changes in the electroencephalographic pattern in male or in female subjects. Our studies indicate the adult human VNO is a functional chemosensory organ with a sexually dimorphic specificity and the ability to transduce signals which modulate certain autonomic parameters.


Low affinity of FMRFamide and four FaRPs (FMRFamide-related peptides), including the mammalian-derived FaRPs F-8-Famide (NPFF) and A-18-Famide, for opioid mu, delta, kappa 1, kappa 2a, or kappa 2b receptors.
The binding affinities at opioid receptor subtypes in rat or guinea pig brain membranes were determined for the neuropeptide FMRFamide (Phe-Met-Arg-Phe-NH2), the two mammalian-derived FMRFamide-related peptides F-8-Famide (NPFF; Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe- NH2) and A-18-Famide (Ala-Gly-Glu-Gly-Leu-Ser-Ser-Pro-Phe-Trp-Ser-Leu-Ala-Ala-Pro-Gln-Arg-Phe -NH2), and the two other FMRFamide-related peptides Tyr-Phe-Met-Arg-Phe-NH2 (Tyr-FMRFamide) and Pro-Gln-Arg-Phe-NH2 (Pro-Gln-RFamide). The mu and delta sites were labeled in rat brain membranes using tritiated [D-Ala2, N-MePhe4,Gly-ol5] enkephalin ([3H]DAMGO) and [D-Ala2,D-Leu5]enkephalin ([3H]DADLE), respectively. The kappa sites were labeled in guinea pig brain using [3H]U-69,593 after treatment with BIT and FIT for kappa 1 and [3H]bremazocine after pretreatment with BIT and FIT for kappa 1 and [3H]bremazocine after pretreatment with BIT and FIT for kappa 2. The kappa 2a binding sites were assayed using [Leu5]enkephalin to block kappa 2b sites and the kappa 2b sites were assayed using (-)-(1S,2S)-U50,488 to block kappa 2a sites. Neither FMRFamide nor any of the FMRFamide-related peptides (up to 61.0 microM) displayed significant affinity at any of the subtypes of opioid receptor. Hence, the known ability of FMRFamide and FaRPs to interact with the opioid system does not appear to be related to direct binding to these opioid receptors.


Caudal Nucleus Accumbens Core Is Critical in the Regulation of Cue-Elicited Approach-Avoidance Decisions.
The nucleus accumbens (NAc) is thought to be a site of integration of positively and negatively valenced information and action selection. Functional differentiation in valence processing has previously been found along the rostrocaudal axis of the shell region of the NAc in assessments of unconditioned motivation. Given that the core region of the NAc has been implicated in the elicitation of motivated behavior in response to conditioned cues, we sought to assess the role of caudal, intermediate, and rostral sites within this subregion in cue-elicited approach-avoidance decisions. Rats were trained to associate visuo-tactile cues with appetitive, aversive, and neutral outcomes. Following the successful acquisition of the cue-outcome associations, rats received microinfusions of GABAA and GABAB receptor agonists (muscimol/baclofen) or saline into the caudal, intermediate, or rostral NAc core and were then exposed to a superimposition of appetitively and aversively valenced cues versus neutral cues in a "conflict test," as well as to the appetitive versus neutral cues, and aversive cues versus neutral cues, in separate conditioned preference/avoidance tests. Disruption of activity in the intermediate to caudal parts of the NAc core resulted in a robust avoidance bias in response to motivationally conflicting cues, as well as a potentiated avoidance of aversive cues as compared with control animals, coupled with an attenuated conditioned preference for the appetitive cue. These results suggest that the caudal NAc core may have the capacity to exert bidirectional control over appetitively and aversively motivated responses to valence signals.


Splitting of second heart sound in patients with aortic regurgitation.
The intervals between the two components of the second heart sound (A2-P2), measured from the phonocardiograms of 50 patients with severe aortic regurgitation, were compared with those of 57 normal subjects. During expiration the second sound was single or closely split in both groups. Inspiratory augmentation of the A2-P2 interval of less than 0.02 sec occurred in one normal subject (2%) and in 42 patients with aortic regurgitation (86%). Thus the great majority of patients with severe aortic regurgitation had either a single or a closely split second heart sound during both phases of respiration. The mean Q-A2 intervals (rate corrected) of 30 patients, in whom severe aortic regurgitation was documented by contrast angiographic studies, were longer than those of normal subjects during both phases of respiration, and the difference was statistically more significant (P < 0.02) during inspiration. Thus prolongation of the Q-A2 interval was the main factor responsible for the disappearance of the normal asynchrony of semilunar valve closure, in patients with aortic regurgitation.


Low-temperature, high-mobility, solution-processed metal oxide semiconductors fabricated with oxygen radical assisted perchlorate aqueous precursors.
In this report, a simple and general chemical route for fabricating MO semiconducting films at a relatively low temperature without any fuel additives or special annealing steps was demonstrated. The precursor, which consisted of perchlorate, nitrate, and water, is easily converted into In2O3 at an annealing temperature of 250 °C due to oxygen radical assisted decomposition and generation of a large amount of heat. It is found that perchlorate salt can decompose and form an oxide film with high quality at a lower temperature when assisted by nitrate salt. The optimized In2O3-TFT fabricated via this precursor exhibits a saturation mobility of 14.5 cm2 V-1 s-1. Furthermore, this approach has been expanded to the fabrication of ZnO films and attained improved performance, indicating its universality.


Influence of sodium hypochlorite on mechanical properties of K3 nickel-titanium rotary instruments.
Several studies have evaluated the influence of various factors on the fracture of nickel-titanium endodontic rotary instruments. The present study analyzed the influence of sodium hypochlorite on flexural fatigue and torsional properties of nickel-titanium endodontic rotary instruments. New files and files previously exposed to sodium hypochlorite were tested for flexural fatigue and for resistance to fracture by twisting. The t test was used to compare the groups for number of cycles, angle of rotation, and maximum torque required to fracture. No statistical difference existed between these groups. Scanning electron microscopy analysis showed no evidence of localized corrosion in files exposed to sodium hypochlorite solution. These results suggest that the exposure to sodium hypochlorite has no influence on resistance to fracture of K3 rotary instruments.


Pain After Percutaneous Irreversible Electroporation of Renal Tumors Is Not Dependent on Tumor Location.
Irreversible electroporation (IRE) is a non-thermal minimally invasive technique that is used to treat small renal masses (SRMs). Prior work has demonstrated greater narcotic requirements after radiofrequency ablation (RFA) for tumors that are closer to body-wall musculature. We hypothesized that pain after IRE is not dependent on tumor location due to the athermal mechanistic action. A retrospective review of 50 consecutive percutaneous IRE and RFA cases was performed from 2013 to 2014. Eight patients were excluded from analysis due to incomplete anesthesia record and/or multiple ablations per session, leaving 21 patients in each group. Data collected included patient age, sex, body mass index, nephrometry score, shortest distance to the closest body-wall muscle, perioperative narcotic use, and patient-reported pain score. Pearson correlation test and multivariable linear regression were used to identify predictors of postoperative pain, with significance set at p = 0.05. There was no difference in the mean distance from tumor edge to the nearest body-wall muscle between IRE and RFA (2.6 cm vs 2.4 cm, p = 0.729, respectively). Total mean perioperative narcotic usage was 20.4 mg after IRE and 26.7 mg after RFA (p = 0.096). Mean postoperative pain score (scale 0-10) was slightly higher after RFA (4.3) compared with IRE (2.4), but this was not statistically significant (p = 0.088). Pearson correlation test identified tumor proximity to be significiantly associated with both pain score (p = 0.011) and postoperative narcotic use (p = 0.049) after RFA but not after IRE. On multivariable analysis, only tumor proximity to the body wall was significantly correlated to pain score (-1.4, p = 0.041) after RFA but was not found to be a factor for pain after IRE. Patients whose tumors lie close to their body-wall musculature do not have greater narcotic requirements or higher pain scores in the perioperative period after IRE. Percutaneous IRE may be preferred over RFA for SRMs that are close to the body wall to minimize pain.


[Diagnosis and therapy of portosystemic encephalopathy].
With respect to the grade of disturbance of consciousness on one hand and concomitant cognitive, psychic and neuromuscular symptoms on the other hand several stages of portosystemic encephalopathy (PSE) are differentiated; moreover, it could recently be demonstrated that a remarkable proportion of cirrhotics without any clinical sign of cerebral dysfunction shows pathological results in psychometric and neurophysiological examinations. Therefore, the stage of subclinical, latent PSE was defined additionally. The assessment of subclinical PSE is usually done using paper-pencil tests or neurophysiological examinations like the EEG. The diagnosis of manifest PSE is made with respect to clinical criteria. Other possible causes of cerebral dysfunction have to be excluded by laboratory, neurological and eventually radiological examination. Therapy of PSE aims at the reduction production and resorption of ammonia. New therapeutic approaches, which have been proposed with regard to pathophysiological aspects of PSE other than ammonia, have not yet been established.


Deconstructing One Medical School's Pain Curriculum: II. Partnering with Medical Students on an Evidence-Guided Redesign.
To reshape medical education about pain to present it as a population-based public health process as well as a neuron-centered phenomenon. Collaborate with students to apply a recent inventory of pain-related preclinical curricular content and clinical training in order to modify the current multiyear presentation and offer a broadened social perspective on pain. Appraise fourth-year medical students' pain-related educational needs by surveying their knowledge, attitudes, experience with the curriculum, and self-reported assessment of pain-related competencies. University-affiliated medical school and its fourth-year medical students. Analysis of a detailed inventory of first- and second-year curricula. Survey of graduating medical students assessing attitudes, skills, and confidence. Construction of a fourth-year pain education elective and collaboration with enrollees to better integrate pain throughout the four-year curriculum. This student-faculty collaboration produced an evidence-guided proposal to reorganize pain-related content across the longitudinal medical curriculum. An attitudes/skills/confidence survey of graduating medical students (104 respondents of 200 polled) found that 70% believed chances for successful outcomes treating chronic pain were low. Self-evaluated competency was high for evaluating (82%) and managing (69%) acute pain; for chronic pain, both were lower (evaluating = 38%; managing = 6%). Self-evaluated knowledge of pain physiology and neurobiology was poor (14%), fair (54%), or good (30%), but rarely excellent (2%). To meet graduating students' desire for increased competency in pain, pain-related curricula can and should be reorganized to include pain as a disease state and a widespread public health burden, not merely a symptom.


Lectin histochemistry of the rat lymph node: visualisation of stroma, blood vessels, sinuses, and macrophages. A contribution to the concept of an immune accessory role of sinus-lining endothelia.
The lectin Chelidonium majus agglutinin (CMA) was previously shown to visualise endothelia of all blood vessels and those lining sinuses of red pulp, stromal reticular meshwok (RM) and dendritic cells of lymphatic follicles in white pulp of the spleen in rats. The aim of the present study was the analysis of CMA and some other lectins in labelling RM, vascular structures and macrophages in lymph nodes of rats. It appeared that CMA stained the entire RM, dendritic cells, lining cells of sinuses and all types of blood vessels. Sinus-lining cells of lymph nodes were labelled with CMA and mannose-, GalNac-, and sialic acid-specific lectins. Moreover, lymph node macrophages were labelled above all by mannose specific lectins. The broad lectin-binding pattern of sinuses--not observed in rat spleen- and CMA-reactivity of both sinus-lining and dendritic cells corroborates the hypothesis that lymph node sinus-lining endothelia are precursors or a special type of immune accessory cells.


A Short Measure of the Revised Reinforcement Sensitivity Theory - RSQ17.
The need for a research and practical tool, such as a short, reliable, and valid personality assessment test, suggests researchers to create shortened versions of original instruments. Reinforcement sensitivity questionnaire (RSQ) was created in line with some basic premises of revised Reinforcement sensitivity theory, which proposes three motivational and emotional systems: Behavioral inhibition system (BIS), responsible for scanning environment for potential threats, Behavioral activation system (BAS), responsible for aproaching behavior, and the Fight/Flight/Freeze system (FFFS), responsible for behavior in the present threat. RSQ comprises five scales: BIS, BAS, Fight, Flight, and Freeze. The aim of this study was to develop a short version of RSQ, which would be beneficial to both research and practical purposes. Item response theory analyses were used for item selection. The study comprised two samples of participants, whereby Sample 1 (N = 837, 34.6% male, aged 18 - 82, M = 31.63, SD = 13.54) served as the derivation sample, while Sample 2 (818 participants, 43.6% male, 18-75 years, M = 29.65, SD = 12.52) served as validation sample. Factorial validity of the short RSQ was examined on both Sample 1 and Sample 2. Convergent and divergent validity of the short RSQ was examined using RST-PQ, Jackson-5, BIS/BAS scales, and Big Five Inventory. The results point to satisfactory internal consistency, factorial validity, and construct validity of the short RSQ, suggesting that it is an adequate measure for research settings or other contexts which require the use of short personality questionnaires.


Externalizing behavior from early childhood to adolescence: Prediction from inhibition, language, parenting, and attachment.
The aim of the current research was to disentangle four theoretically sound models of externalizing behavior etiology (i.e., attachment, language, inhibition, and parenting) by testing their relation with behavioral trajectories from early childhood to adolescence. The aim was achieved through a 10-year prospective longitudinal study conducted over five waves with 111 referred children aged 3 to 5 years at the onset of the study. Clinical referral was primarily based on externalizing behavior. A multimethod (questionnaires, testing, and observations) approach was used to estimate the four predictors in early childhood. In line with previous studies, the results show a significant decrease of externalizing behavior from early childhood to adolescence. The decline was negatively related to mothers' coercive parenting and positively related to attachment security in early childhood, but not related to inhibition and language. The study has implications for research into externalizing behavior etiology recommending to gather hypotheses from various theoretically sound models to put them into competition with one another. The study also has implications for clinical practice by providing clear indications for prevention and early intervention.


Investigational BET bromodomain protein inhibitors in early stage clinical trials for acute myelogenous leukemia (AML).
Acute myelogenous leukemia (AML) is a heterogeneous group of malignancies driven by genetic mutations and deregulated epigenetic control. Relapse/refractory disease remains frequent in younger patients and even more so in older patients, including treatment with epigenetic drugs in this age group, mainly with hypomethylating agents. New treatment strategies are urgently needed. The recent discovery that epigenetic readers of the bromodomain (BRD) and extraterminal (BET) protein family, are crucial for AML maintenance by transcription of oncogenic c-MYC lead to rapid development of BET inhibitors entering clinical trials. Areas covered: We provide a critical overview using main sources for the use of BET inhibitors in AML treatment. Limits of this treatment approach including resistance mechanisms and future directions including development of new generation BET inhibitors and combination strategies with other drugs are detailed. Expert opinion: BET inhibitors were expected to overcome limits of conventional treatment in patients as impressive in vitro data emerged recently in well-characterized AML subsets, including those associated with poor risk characteristics in the clinic. Nevertheless single activity of BET inhibitors appears to be modest and resistance mechanisms were already identified. BET inhibitors with alternative mechanisms of action and/or combination strategies with epigenetic drugs should be tested.


Qualitative dissimilarities of insulin and proinsulin binding and action in vitro at IM 9-lymphocytes.
To prove hormone analogy of insulin and proinsulin 1. biochemical characterization of binding, 2. competition-inhibition-assay, 3. up-or-down-regulation, and 4. 3H-thymidine incorporation in IM 9-lymphocytes in vitro were applied. Marked differences in binding and biological activity of insulin and proinsulin in the physiological range (10 pM-1 nM) are obvious. Heterogeneity of specific insulin and proinsulin binding sites is suggested.


Progressive swelling and radiopaque mass in maxillary sinus: formation of stone.
Although radiopaque lesions located in the maxillary sinus are rare, differential diagnosis should include a number of pathologies. Formation of stone, namely "antrolith" in the paranasal sinuses is a very rare phenomenon and it should be considered in the differential diagnosis, as it is usually located in the maxillary sinus. In this article, we present a 47-year-old male case with unilateral chronic sinusitis for a long time and calcification in maxillary sinus in the light of clinical/radiographic findings of the lesion and treatment strategy.


Routine management of volume status after aneurysmal subarachnoid hemorrhage.
Prophylactic use of hypervolemia and hypertension is believed to present an option to decrease the incidence of symptomatic vasospasm after aneurysmal subarachnoid hemorrhage and improve neurologic outcome. A Medline literature search was conducted to review available evidence regarding volume management after subarachnoid hemorrhage. Quality of selected studies was evaluated, using the standardized GRADE system. Eleven studies focused on prophylactic hypervolemic therapy after aneurysmal subarachnoid hemorrhage were identified, including four randomized controlled trials. Available studies showed a large heterogeneity in physiologic treatment goals and interventions applied. The oldest and smallest randomized controlled trial suggested a positive effect, but had severe limitations in trial design. Neither of the other randomized controlled studies showed outcome benefit with hypervolemic therapy. Results from observational studies were not found to support the use of prophylactic hypervolemia and hypertension. Complication frequency was repeatedly reported to be higher with the application of prophylactic hypervolemia. In summary, prophylactic hyperdynamic therapy after subarachnoid hemorrhage has not been adequately shown to effectively raise cerebral blood flow or improve neurological outcome. In contrast, there is evidence for harm using overly aggressive hydration.


Rubella virus capsid protein: a small protein with big functions.
Virus replication occurs in the midst of a life or death struggle between the virus and the infected host cell. To limit virus replication, host cells can activate a number of antiviral pathways, the most drastic of which is programmed cell death. Whereas large DNA viruses have the luxury of encoding accessory proteins whose main function is to interfere with host cell defences, the genomes of RNA viruses are not large enough to encode proteins of this type. Recent studies have revealed that proteins encoded by RNA viruses often play multiple roles in the battles between viruses and host cells. In this article, we discuss the many functions of the rubella virus capsid protein. This protein has well-defined roles in virus assembly, but recent research suggests that it also functions to modulate virus replication and block host cell defences.


Age - Does it really count? A study of the Perioperative Morbidity and Long-Term Outcome in Patients Above 70 Years of Age Undergoing Spine surgery for Lumbar Degenerative Disorders.
Objective To assess the long-term outcome and perioperative morbidity in spine surgeries for lumbar degenerative disorders and, thereby, to evaluate the safety of surgery in the aging population. Methods Retrospective study of patients aged > 70 years, operated for degenerative lumbar disorders between 2011 and 2015. We evaluated patient demographic, clinical and surgical data; comorbidities, perioperative complications, pre & postoperative pain scores and Oswestry disability index (ODI) scores, patient satisfaction and overall mortality. Results A total of 103 patients (Males: Females55:48) with mean age 74.6 years (70-85yrs) were studied. 60 patients (58.2%) had decompression alone, while 43 (41.8%) had decompression & fusion. Mean hospital stay was 5.7days. Mean follow-up was 47.6months (24-73mnths). Patients reported significant improvement in backpain (Numerical pain score 7.7 vs 1.6; p < 0.001), leg pain (Numerical pain score 7.4 vs 1.7; p < 0.001), disability (ODI 82.3 vs 19.1; p < 0.001) and walking distance ( p < 0.001). 76% patients were satisfied with the results at the time of final follow-up. 26 patients (25.24%) had perioperative complications which were all minor, without mortality. Most common intraoperative & postoperative complications were dural tear (6.79%) & urinary tract infection (6.79%) respectively. Conclusions With meticulous perioperative care lumbar spine surgery is safe and effective in elderly population. Patients had longer mean hospital stay in view of the gradual and comprehensive rehabilitation program. Presence of comorbidities or minor perioperative complications did not increase the overall morbidity or affect the clinical outcomes of surgery in our study.


Peptides corresponding to a predictive alpha-helical domain of human immunodeficiency virus type 1 gp41 are potent inhibitors of virus infection.
To define the role of the human immunodeficiency virus type 1 (HIV-1) envelope proteins in virus infection, a series of peptides were synthesized based on various regions of the HIV-1 transmembrane protein gp41. One of these peptides, DP-178, corresponding to a region predictive of alpha-helical secondary structure (residues 643-678 of the HIV-1LAI isolate), has been identified as a potent antiviral agent. This peptide consistently blocked 100% of virus-mediated cell-cell fusion at < 5 ng/ml (IC90 approximately 1.5 ng/ml) and gave an approximately 10 times reduction in infectious titer of cell-free virus at approximately 80 ng/ml. The inhibitory activity was observed at peptide concentrations approximately 10(4) to 10(5) times lower than those at which cytotoxicity and cytostasis were detected. Peptide-mediated inhibition is HIV-1 specific in that approximately 10(2) to 10(3) times more peptide was required for inhibition of a human immunodeficiency virus type 2 isolate. Further experiments showed that DP-178 exhibited antiviral activity against both prototypic and primary HIV-1 isolates. As shown by PCR analysis of newly synthesized proviral DNA, DP-178 blocks an early step in the virus life cycle prior to reverse transcription. Finally, we discuss possible mechanisms by which DP-178 may exert its inhibitory activity.


Binding of alpha-fetoprotein by immobilized monoclonal antibodies during episodes of zero-gravity obtained by parabolic flight.
Alpha-fetoprotein (AFP), a single chain polypeptide which is synthesized by the liver and yolk sac of the human fetus, provided a model ligand for assessing the effects of microgravity on ligand binding to surface-immobilized model receptor molecules. Monoclonal antibodies, used as receptors for AFP, were immobilized by covalent attachment to latex microparticles. Zero gravity environment was obtained by parabolic flight aboard NASA 930, a modified KC-135 aircraft. During the onset of an episode of zero gravity, ligand and receptor were mixed. Timed incubation (20 s) was terminated by centrifugation, the supernatant removed, and microparticles were assessed for bound AFP by immunochemical methods. The extent of binding was not influenced by microgravity, when compared with 1-G controls, which suggests that aberrant cellular activities observed in microgravity are not the simple expression of altered macromolecular interactions.


Reduction of blood pressure variability by combination therapy in spontaneously hypertensive rats.
To test the hypothesis that combination therapy might be a powerful tool to reduce blood pressure variability (BPV) in the treatment of hypertension. In chronic studies, male spontaneously hypertensive rats were given drugs in their rat chow for 18 weeks (n = 9 or 10 for each group of rats). Doses were as follows: atenolol (10 mg/kg per day), amlodipine (1 mg/kg per day), a combination of atenolol and amlodipine (2.5 + 0.25, 5 + 0.5, and 10 + 1 mg/kg per day), hydrochlorothiazide (8 mg/kg per day), enalapril (3.2 mg/kg per day), and a combination of hydrochlorothiazide and enalapril (2 + 0.8, 4 + 1.6, and 8 + 3.2 mg/kg per day). In acute studies, drugs were perfused through the left femoral vein in conscious rats. In chronic studies, compared with monotherapy, the combinations of two antihypertensive drugs were more effective in reducing the blood pressure (BP), BPV, and organ damage. The indexes of organ damage were all positively related to BP and/or BPV. In acute studies, the constant infusion of phenylephrine (6.25 microg/kg per min) markedly increased the BP, but showed no significant effects on BPV. The infusion of a combination of atenolol and amlodipine (62.5 + 6.25 microg/kg per min) or a combination of hydrochlorothiazide and enalapril (500 + 200 microg/kg per min) significantly reduced the BP and BPV; moreover, a significant reduction in BPV was still found when the rat's BP was restored to control levels by a concomitant infusion of phenylephrine. Combination therapy may be a powerful and useful tool for BPV reduction in the treatment of hypertension. In addition to the BP reduction, the decrease in BPV may contribute significantly to the prevention of organ damage in hypertension.


Merging Ion Concentration Polarization between Juxtaposed Ion Exchange Membranes to Block the Propagation of the Polarization Zone.
The ion concentration polarization (ICP) phenomenon is one of the most prevailing methods to preconcentrate low-abundance biological samples. The ICP induces a noninvasive region for charged biomolecules (i.e., the ion depletion zone), and targets can be preconcentrated on this region boundary. Despite the high preconcentration performances with ICP, it is difficult to find the operating conditions of non-propagating ion depletion zones. To overcome this narrow operating window, we recently developed a new platform for spatiotemporally fixed preconcentration. Unlike preceding methods that only use ion depletion, this platform also uses the opposite polarity of the ICP (i.e., ion enrichment) to stop the propagation of the ion depletion zone. By confronting the enrichment zone with the depletion zone, the two zones merge together and stop. In this paper, we describe a detailed experimental protocol to build this spatiotemporally defined ICP platform and characterize the preconcentration dynamics of the new platform by comparing them with those of the conventional device. Qualitative ion concentration profiles and current-time responses successfully capture the different dynamics between the merged ICP and the stand-alone ICP. In contrast to the conventional one that can fix the preconcentration location at only ~5 V, the new platform can produce a target-condensed plug at a specific location in the broad ranges of operating conditions: voltage (0.5-100 V), ionic strength (1-100 mM), and pH (3.7-10.3).


A survey of tobacco dependence treatment guidelines in 121 countries.
To report progress among Parties to the World Health Organization (WHO) Framework Convention on Tobacco Control (FCTC) in developing national tobacco treatment guidelines in accordance with FCTC Article 14 guideline recommendations. Cross-sectional study. Electronic survey from December 2011 to August 2012; participants were asked to complete either an online or attached Microsoft Word questionnaire. One hundred and sixty-three of the 173 Parties to the FCTC at the time of our survey. The 51-item questionnaire contained 30 items specifically on guidelines. Questions covered the areas of guidelines writing process, content, key recommendations and other characteristics. One hundred and twenty-one countries (73%) responded. Fifty-three countries (44%) had guidelines, ranging from 75% among high-income countries to 11% among low-income countries. Nearly all guidelines recommended brief advice (93%), intensive specialist support (93%) and medications (96%), while 66% recommended quitlines. Fifty-seven percent had a dissemination strategy, 76% stated funding source and 68% had professional endorsement. Fewer than half of the Parties to the WHO FCTC have developed national tobacco treatment guidelines, but, where guidelines exist, they broadly follow FCTC Article 14 guideline recommendations.


In vitro permeability of etch-and-rinse and self-etch adhesives used for immediate dentin sealing.
To investigate the permeability of deep dentin following immediate sealing with different etch-and-rinse and self-etch adhesives (Single Bond 2, Adper Prompt L-Pop, Clearfil Protect Bond, Clearfil S3 Bond, G-Bond) and a dentin desensitizer (Gluma). Fluid-transport model was used to measure fluid conductance during and after application of adhesives. Polyvinylsiloxane impressions of bonded dentin were taken to monitor fluid transudation from the surface of the adhesives. The area and number of dentinal fluid droplets and/or blisters were calculated using image analysis. None of the adhesives were able to block fluid conductance completely. The fluid conductance values of the adhesives displayed the following statistical ranking (p<0.05): G-Bond≤Clearfil Protect Bond<Smear-layer-covered dentin<S3 Bond≤Single Bond 2≤Adper Prompt L-Pop<Gluma Desensitizer<Acid-etched dentin. Highly significant correlation was observed between the permeability of the tested adhesives and the area fraction of fluid droplets/blisters on the adhesive surfaces (r=0.99, p<0.01).


Gallic Acid Content and an Antioxidant Mechanism Are Responsible for the Antiproliferative Activity of 'Ataulfo' Mango Peel on LS180 Cells.
Mango "Ataulfo" peel is a rich source of polyphenols (PP), with antioxidant and anti-cancer properties; however, it is unknown whether such antiproliferative activity is related to PP's antioxidant activity. The content (HPLC-DAD), antioxidant (DPPH, FRAP, ORAC), and antiproliferative activities (MTT) of free (FP) and chemically-released PP from mango 'Ataulfo' peel after alkaline (AKP) and acid (AP) hydrolysis, were evaluated. AKP fraction was higher (µg/g DW) in gallic acid (GA; 23,816 ± 284) than AP (5610 ± 8) of FR (not detected) fractions. AKP fraction and GA showed the highest antioxidant activity (DPPH/FRAP/ORAC) and GA's antioxidant activity follows a single electron transfer (SET) mechanism. AKP and GA also showed the best antiproliferative activity against human colon adenocarcinoma cells (LS180; IC50 (µg/mL) 138.2 ± 2.5 and 45.7 ± 5.2) and mouse connective cells (L929; 93.5 ± 7.7 and 65.3 ± 1.2); Cheminformatics confirmed the hydrophilic nature (LogP, 0.6) and a good absorption capacity (75%) for GA. Data suggests that GA's antiproliferative activity appears to be related to its antioxidant mechanism, although other mechanisms after its absorption could also be involved.


Induction of interleukin 1 by Legionella pneumophila in murine peritoneal macrophage cultures.
Legionella pneumophila-induced production of both membrane-associated and secreted interleukin 1 (mIL-1 and sIL-1, respectively) was examined utilizing peritoneal macrophages from BALB/c mice. The Legionella preparations for these studies included viable bacteria and formalin-killed whole cell preparations. Both of the preparations induced mIL-1 and sIL-1 in a dose-dependent fashion. However, the viable bacteria required about 1 log lower concentrations than the formalin-killed bacteria to induce the same level of IL-1 activity measured in the thymocyte proliferation assay. Kinetic studies showed that mIL-1 and sIL-1 were detectable within 4 hr after addition of either of the L. pneumophila preparations to the peritoneal macrophage cultures, with peak levels achieved within 24 hr. These results indicate that L. pneumophila is a potent inducer of both mIL-1 and sIL-1 in normal mouse peritoneal macrophage cultures.


Rational optimization of a bispecific ligand trap targeting EGF receptor family ligands.
The human epidermal growth factor (EGF) receptor (HER) family members cooperate in malignancy. Of this family, HER2 does not bind growth factors and HER3 does not encode an active tyrosine kinase. This diversity creates difficulty in creating pan-specific therapeutic HER family inhibitors. We have identified single amino acid changes in epidermal growth factor receptor (EGFR) and HER3 which create high affinity sequestration of the cognate ligands, and may be used as receptor decoys to downregulate aberrant HER family activity. In silico modeling and high throughput mutagenesis were utilized to identify receptor mutants with very high ligand binding activity. A single mutation (T15S; EGFR subdomain I) enhanced affinity for EGF (two-fold), TGF-alpha (twenty-six-fold), and heparin-binding (HB)-EGF (six-fold). This indicates that T15 is an important, previously undescribed, negative regulatory amino acid for EGFR ligand binding. Another mutation (Y246A; HER 3 subdomain II) enhanced neuregulin (NRG)1-beta binding eight-fold, probably by interfering with subdomain II-IV interactions. Further work revealed that the HER3 subunit of an EGFR:HER3 heterodimer suppresses EGFR ligand binding. Optimization required reversing this suppression by mutation of the EGFR tether domain (G564A; subdomain IV). This mutation resulted in enhanced ligand binding (EGF, ten-fold; TGF-alpha, thirty-four-fold; HB-EGF, seventeen-fold; NRG1-beta, thirty-one-fold). This increased ligand binding was reflected in improved inhibition of in vitro tumor cell proliferation and tumor suppression in a human non-small cell lung cancer xenograft model. In conclusion, amino acid substitutions were identified in the EGFR and HER3 ECDs that enhance ligand affinity, potentially enabling a pan-specific therapeutic approach for downregulating the HER family in cancer.


Sonchus yellow net virus core particles form on ring-like nuclear structure enriched in viral phosphoprotein.
The phosphoprotein (P) of the nucleorhabdovirus sonchus yellow net virus has been shown to accumulate in ring-shaped structures in virus-infected nuclei. Further examination by live-cell imaging, in combination with structural examination by transmission electron microscopy and immunolocalization demonstrated that P-rings do not form in association with nucleoli. Furthermore, viral cores were shown to condense on the nucleoplasm-contacting surface of the rings. The data presented here offer evidence for the site of nucleocapsid assembly in SYNV-infected nuclei.


An unexpected thermal-ring-rearrangement of benzochromenes to inden-3-yl-naphthols with pTsOH.
Described here is the first report of an unexpected thermal-ring rearrangement (TRR) of benzochromenes to indene derivatives promoted by pTsOH. This cascade ring-rearrangement proceeds through the protonation of benzochromenes by an acid catalyst followed by ring-opening and ring-closure by an intramolecular Friedel-Crafts cyclization to provide a new bicyclic framework, inden-3-yl-naphthols bearing a quaternary center, which also exhibited atropisomerism. Regioselectivity, broad substrate scope, high yields, solvent-free conditions and atom economy are the additional high points of this ring-rearrangement.


Teleoncology for Veterans: High Patient Satisfaction Coupled With Positive Financial and Environmental Impacts.
There was rapid adoption of teleoncology care in the Veterans Health Administration during the COVID-19 pandemic. One third of 9 million Veterans Health Administration enrolled Veterans live in rural areas. Although digital solutions can expand capacity, enhance care access, and reduce financial burden, they may also exacerbate rural-urban health disparities. Careful evaluation of patients' perceptions and policy tradeoffs are necessary to optimize teleoncology postpandemic. Patients with ≥ 1 teleoncology visit with medical, surgical, or radiation oncology between March 2020 and June 2020 were identified retrospectively. Validated, Likert-type survey assessing patient satisfaction was developed. Follow-up survey was conducted on patients with ≥ 1 teleoncology visit from August 2020 to January 2021. Travel distance, time, cost, and carbon dioxide emissions were calculated based on zip codes. A hundred surveys were completed (response rate, 62%). Patients overall were satisfied with teleoncology (83% Agree or Strongly Agree) but felt less satisfied than in-person visits (47% Agree or Strongly Agree). Audiovisual component improved patient perception of involvement in care, ability to self-manage health or medical needs, and comparability to in-person visits. Follow-up survey demonstrated similar satisfaction. Total travel-related savings are as follows: 86,470 miles, 84,374 minutes, $49,720 US dollars, and 35.5 metric tons of carbon dioxide. Veterans are broadly satisfied with teleoncology. Audiovisual capabilities are critical to satisfaction. This is challenging for rural populations with lack of technology access. Patients experienced financial and time savings, and society benefitted from reduced carbon emissions. Continued optimization is needed to enhance patient experience and address secondary effects.


Components of mesophyll resistance and their environmental responses: A theoretical modelling analysis.
Mesophyll resistance (rm ), stomatal resistance, and biochemical limitations are recognized as three critical factors limiting leaf photosynthesis. Contrary to the expectation of being a constant, rm not only varies with light and CO2 conditions but also shows different responses among species. To elucidate the mechanistic basis of these responses, we derived an analytical model of rm , which incorporates various anatomical and biochemical factors including permeabilities of cell wall and chloroplast envelope to CO2 and HCO3- , carbonic anhydrase activities in cytosol and stroma, Rubisco activities, and relative location of mitochondria and chloroplast. The robustness of this model was confirmed by comparing the predicted rm and its components to numerical models developed at cell and leaf levels, which incorporate detailed 3-dimensional cell and leaf anatomies, CO2 hydration and diffusion processes from intercellular air space to stroma, and CO2 fixation by Rubisco. A combination of these model analyses shows that the varying rm is influenced by four biochemical factors: (a) nonuniform photosynthesis status across the leaf, (b) photorespiration and respiration, (c) bicarbonate leakage on the chloroplast envelope, and (d) hydration activity in cytosol and stroma. This study provides a theoretical framework to study components of rm and their responses to environmental perturbations.


DNA in extracellular vesicles: from evolution to its current application in health and disease.
Extracellular vesicle (EV) secretion is a highly conserved evolutionary trait in all organisms in the three domains of life. The packaging and release of EVs appears to be a bulk-flow process which takes place mainly under extreme conditions. EVs participate in horizontal gene transfer, which supports the survival of prokaryotic and eukaryotic microbes. In higher eukaryotes, almost all cells secrete a heterogeneous population of EVs loaded with various biomolecules. EV secretion is typically higher in cancer microenvironments, promoting tumor progression and metastasis. EVs are now recognized as additional mediators of autocrine and paracrine communication in health and disease. In this context, proteins and RNAs have been studied the most, but extracellular vesicle DNA (EV-DNA) has started to gain in importance in the last few years. In this review, we summarize new findings related to the loading mechanism(s), localization, and post-shedding function of EV-DNA. We also discuss the feasibility of using EV-DNA as a biomarker when performing a liquid biopsy, at the same time emphasizing the lack of data from clinical trials in this regard. Finally, we outline the potential of EV-DNA uptake and its interaction with the host genome as a promising tool for understanding the mechanisms of cancer evolution.


[Application of the lateral preputial fascial island flap in hypospadias reoperation].
To evaluate the application and effect of the lateral preputial fascial island flap (LPFIF) in hypospadias reoperation. We retrospectively analyzed the clinical data on the children patients undergoing hypospadias reoperation with LPFIF in our Department of Urology from December 2016 to June 2019. A total of 85 cases were included in this study, including 18 cases of LPFIF, 19 cases of Duplay technique, 25 cases of tubularized incised plate urethroplasty (TIP) and 23 cases of Mathieu urethroplasty. The patients were aged from 18 months to 12 years and 8 months, averaging 4.6 years. Postoperatively, all the children were followed up for ≥ 6 months, which found satisfactory appearance of the penis body and glans, the urethral orifice in the normal position of the glans, and the external orifice of the urethra fissured. Urethral fistula occurred in 2 cases and glans dehiscence in 1 after LPFIF; 5 of the patients presented urethral fistula after Duplay; 3 developed urethral fistula and 1 urethral stricture after TIP; 6 showed urethral fistula and 2 glans dehiscence after Mathieu urethroplasty. No postoperative urethral stricture, urethral diverticulum or flap necrosis occurred in any of the cases. Hypospadias reoperation succeeded in 83.3% (15/18) of the cases after LPFIF. The urine flow curve of the LPFIF cases was bell-shaped or high flat-shaped, with a maximum urinary flow rate of 8.56 ± 3.99 ml/s and an average urinary flow rate of 5.23 ± 2.32 ml/s, not significantly different from those of the TIP and Duplay cases. There is no statistically significant difference in the success rate of hypospadias reoperation between TIP and LPFIF. Urethroplasty with LPFIF can be used as one of the surgical options for hypospadias reoperation.


GRAPPA 2022 Trainee Symposium: A Summary of Oral and Poster Presentations.
One of the highlights of the 2022 Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA) annual meeting was the trainee symposium. Dermatology and rheumatology trainees presented their research related to psoriasis and psoriatic arthritis. This report briefly reviews 5 oral presentations and 15 posters that were selected for this annual meeting. Topics include basic/translational, clinical, and outcomes research reflecting the spectrum of GRAPPA's effort and influence nationally and internationally in the area of psoriatic diseases.


Facile fabrication of biodegradable endothelium-mimicking coatings on bioabsorbable zinc-alloy stents by one-step electrophoretic deposition.
The zinc-alloy stent is one of the best potential candidates for bioabsorbable metal stents because of its appropriate corrosion rate aligned to the duration of the healing process of the surrounding vessel tissues. However, excessive release of zinc ions, causing cytotoxicity of endothelial cells, and insufficient surface bio-functions of Zn-alloy stents lead to considerable challenge in their application. Herein, one-step electrophoretic deposition was employed to apply a hybrid coating of polycarbonate, tannic acid, and copper ions with tailored functions on Zn-alloy stents to enhance their corrosion resistance and provide an endothelium-mimicking surface. Specifically, the synthesized amino-functionalized aliphatic polycarbonates endowed the hybrid coating with specific surface-erosion properties, resulting in superior corrosion resistance and long-term stability in degradation tests both in vitro and in vivo. The immobilized copper ions enabled the catalytic generation of nitric oxide and promoted the adhesion and proliferation of endothelial cells on zinc alloy. The added tannic acid firmly chelated the copper ions and formed durable phenolic-copper-amine crosslinked networks by electrostatic interaction, resulting in long-term stability of the hybrid coating during the 21 day dynamic immersion test. Tannic acid exerted a synergistic antibacterial effect with copper ions as well as a reduction in the inflammatory response to the zinc substrate. In addition, the hybrid coating improved the in vitro hemocompatibility of zinc alloys. By adjusting the amount of chelated copper in the coating system, the biological function of the corresponding coatings can be controlled, providing a facile surface treatment strategy to promote the progress of zinc-alloy stents in clinical applications.


Association of DNA-Methylation Profiles With Immune Responses Elicited in Breast Cancer Patients Immunized With a Carbohydrate-Mimicking Peptide: A Pilot Study.
Immune response to a given antigen, particularly in cancer patients, is complex and is controlled by various genetic and environmental factors. Identifying biomarkers that can predict robust response to immunization is an urgent need in clinical cancer vaccine development. Given the involvement of DNA methylation in the development of lymphocytes, tumorigenicity and tumor progression, we aimed to analyze pre-vaccination DNA methylation profiles of peripheral blood mononuclear cells (PBMCs) from breast cancer subjects vaccinated with a novel peptide-based vaccine referred to as P10s-PADRE. This pilot study was performed to evaluate whether signatures of differentially methylated (DM) loci can be developed as potential predictive biomarkers for prescreening subjects with cancer who will most likely generate an immune response to the vaccine. Genomic DNA was isolated from PBMCs of eight vaccinated subjects, and their DNA methylation profiles were determined using Infinium® MethylationEPIC BeadChip array from Illumina. A linear regression model was applied to identify loci that were differentially methylated with respect to anti-peptide antibody titers and with IFN-γ production. The data were summarized using unsupervised-learning methods: hierarchical clustering and principal-component analysis. Pathways and networks involved were predicted by Ingenuity Pathway Analysis. We observed that the profile of DM loci separated subjects in regards to the levels of immune responses. Canonical pathways and networks related to metabolic and immunological functions were found to be involved. The data suggest that it is feasible to correlate methylation signatures in pre-treatment PBMCs with immune responses post-treatment in cancer patients going through standard-of-care chemotherapy. Larger and prospective studies that focus on DM loci in PBMCs is warranted to develop pre-screening biomarkers before BC vaccination. Clinical Trial Registration: www.ClinicalTrials.gov, Identifier: NCT02229084.


[A case of far-advanced colon cancer with extraluminal progression that required urgent surgical treatment].
A 60-year-old man was admitted to our hospital because of right flank pain. An irregular cystic mass region with calcification was detected in his right abdomen on computed tomography( CT). On the fifth day after admission, the patient's abdominal pain suddenly worsened. Based on a diagnosis of panperitonitis, we performed an urgent laparotomy on the same day. A bulky tumor involving the right colon, duodenum, and retroperitoneum was found in his abdominal cavity. We performed right hemicolectomy and partial resection of the duodenum but could not completely excise the tumor. The manipulation caused a serious duodenal injury; and therefore, pyloric antrum transection, gastrojejunostomy, and cholecystostomy were performed to reduce the inflow of gastric content and bile. Histological examination of the surgical specimens revealed a moderately differentiated adenocarcinoma that originated from the ascending colon. Many cancer cells were detected in the excised margin; thus, the surgical maneuver of choice was absolute noncurative resection. If peritonitis had not been observed, preoperative chemotherapy would have been more appropriate.


Circulating adiponectin levels, body composition and obesity-related variables in Prader-Willi syndrome: comparison with obese subjects.
People with obesity and/or the metabolic syndrome have an increased risk for developing diabetes and cardiovascular disease and may have low adiponectin levels. The obesity associated with Prader-Willi syndrome (PWS) would be expected to have similar complications. However, it was recently reported that, despite their adiposity, people with PWS have reduced visceral fat and are less likely to develop diabetes mellitus or the metabolic syndrome compared with people with simple obesity. To determine if plasma adiponectin levels and other variables relevant to diabetes and cardiovascular risk are different in a cohort of PWS subjects with known genetic subtypes compared with age-, sex- and weight-matched control subjects. Fasting plasma glucose, C-peptide, triglycerides, leptin and cholesterol levels were similar in PWS and obese subjects. Our 20 PWS subjects (mean age = 27.7 years) had higher percent body fat (54.1 vs 48.5%) determined by DEXA measurements and lower percent lean mass (45.9 vs 51.5%) compared with 14 obese controls (mean age = 26.9 year). Plasma adiponectin levels were significantly higher in PWS (15.5 +/- 8.2 microg/ml) than in obese controls (7.5 +/- 2.7 microg/ml). A significant positive correlation was found with insulin sensitivity in PWS subjects (r = 0.75, P = 0.0003) but not in obese controls (r = 0.36, P = 0.20). Our study confirmed an earlier observation of higher adiponectin levels in PWS subjects and less insulin resistance proportionate to their obesity status than found in subjects with simple obesity. Furthermore, no differences were seen in PWS subjects with the chromosome 15 deletion or maternal disomy 15. The reported excessive visceral adiposity in subjects with simple obesity compared with PWS may be associated with decreased production and lower circulating levels of adiponectin.


Community ecology and disease risk: lizards, squirrels, and the Lyme disease spirochete in California, USA.
Vector-borne zoonotic diseases are often maintained in complex transmission cycles involving multiple vertebrate hosts and their arthropod vectors. In the state of California, U.S.A., the spirochete Borrelia burgdorferi, which causes Lyme disease, is transmitted between vertebrate hosts by the western black-legged tick, Ixodes pacificus. Several mammalian species serve as reservoir hosts of the spirochete, but levels of tick infestation, reservoir competence, and Borrelia-infection prevalence vary widely among such hosts. Here, we model the host (lizards, Peromyscus mice, Californian meadow voles, dusky-footed wood rats, and western gray squirrels), vector, and pathogen community of oak woodlands in northwestern California to determine the relative importance of different tick hosts. Observed infection prevalence of B. burgdorferi in host-seeking I. pacificus nymphs was 1.8-5.3%, and our host-community model estimated an infection prevalence of 1.6-2.2%. The western gray squirrel (Sciurus griseus) was the only source of infected nymphs. Lizards, which are refractory to Borrelia infection, are important in feeding subadult ticks but reduce disease risk (nymphal infection prevalence). Species identity is therefore critical in understanding and determining the local disease ecology.


Effect of resveratrol on blood pressure: A systematic review and meta-analysis of randomized, controlled, clinical trials.
Introduction: Results of previous clinical trials evaluating the effect of resveratrol supplementation on blood pressure (BP) are controversial. Purpose: We aimed to assess the impact of resveratrol on BP through systematic review of literature and meta-analysis of available randomized, controlled clinical trials (RCTs). Methods: Literature search included SCOPUS, PubMed-Medline, ISI Web of Science and Google Scholar databases up to 17th October 2017 to identify RCTs investigating the impact of resveratrol on BP. Two review authors independently extracted data on study characteristics, methods and outcomes. Overall, the impact of resveratrol on BP was reported in 17 trials. Results: Administration of resveratrol did not significantly affect neither systolic BP [weighted mean difference (WMD): -2.5 95% CI:(-5.5, 0.6) mmHg; p=0.116; I2=62.1%], nor diastolic BP [WMD: -0.5 95% CI:(-2.2, 1.3) mmHg; p=0.613; I2=50.8], nor mean BP [MAP; WMD: -1.3 95% CI:(-2.8, 0.1) mmHg; p=0.070; I2=39.5%] nor pulse pressure [PP; WMD: -0.9 95% CI:(-3.1, 1.4) mmHg; p=0.449; I2=19.2%]. However, significant WMDs were detected in subsets of studies categorized according to high resveratrol daily dosage (≥300 mg/day) and presence of diabetes. Meta-regression analysis revealed a positive association between systolic BP-lowering resveratrol activity (slope: 1.99; 95% CI: 0.05, 3.93; two-tailed p= 0.04) and Body Mass Index (BMI) at baseline, while no association was detected neither between baseline BMI and MAP-lowering resveratrol activity (slope: 1.35; 95% CI: -0.22, 2.91; two-tailed p= 0.09) nor between baseline BMI and PP-lowering resveratrol activity (slope: 1.03; 95% CI: -1.33, 3.39; two-tailed p= 0.39). Resveratrol was fairly well-tolerated and no serious adverse events occurred among most of the eligible trials. Conclusion: The favourable effect of resveratrol emerging from the current meta-analysis suggests the possible use of this nutraceutical as active compound in order to promote cardiovascular health, mostly when used in high daily dose (≥300 mg/day) and in diabetic patients.


Herpes simplex virus immediate-early promoters are responsive to virus and cell trans-acting factors.
The promoters for each of the immediate-early genes from herpes simplex virus type 1 were cloned and fused to a chloramphenicol acetyltransferase cassette. These chimeric genes were used as targets in a transient expression assay to determine how the immediate-early gene products ICP4 and ICP0 and the virion-associated stimulatory protein Vmw65 affected their expression in HeLa and Vero cells. The basal level of expression from these cassettes differed significantly depending on the extent of 5'-flanking sequence and the cell line that served as host. The promoters from IE-4 and IE-0 behaved in a qualitatively similar fashion independent of the host cell. However, the promoter for ICP27 had a unique response pattern: in Vero cells it acted as an alpha gene promoter, whereas in HeLa cells its response was more like that of a beta gene promoter. The promoter sequences for ICP22 and ICP47 behaved as the IE-4 and IE-0 promoters did in HeLa cells, but their response to the effector molecules in Vero cells was unlike that of other alpha gene promoters we have studied. Evidence is also presented for a role for ICP27 in autoregulation.


Differentiation and Characterization of Cystic Fibrosis Transmembrane Conductance Regulator Knockout Human Pluripotent Stem Cells into Salivary Gland Epithelial Progenitors.
The differentiation of pluripotent stem cells has been used to study disease mechanisms and development. We previously described a method for differentiating human pluripotent stem cells (hPSCs) into salivary gland epithelial progenitors (SGEPs). Here, cystic fibrosis transmembrane conductance regulator (CFTR) knockout hPSCs were differentiated into SGEPs derived from CFTR knockout hESCs (CF-SGEPs) using the same protocol to investigate whether the hPSC-derived SGEPs can model the characteristics of CF. CF-a disease that affects salivary gland (SG) function-is caused by mutations of the CFTR gene. Firstly, we successfully generated CFTR knockout hPSCs with reduced CFTR protein expression using the CRISPR-Cas9 system. After 16 days of differentiation, the protein expression of CFTR decreased in SGEPs derived from CFTR knockout hESCs (CF-SGEPs). RNA-Seq revealed that multiple genes modulating SG development and function were down-regulated, and positive regulators of inflammation were up-regulated in CF-SGEPs, correlating with the salivary phenotype of CF patients. These results demonstrated that CFTR suppression disrupted the differentiation of hPSC-derived SGEPs, which modeled the SG development of CF patients. In summary, this study not only proved that the hPSC-derived SGEPs could serve as manipulable and readily accessible cell models for the study of SG developmental diseases but also opened up new avenues for the study of the CF mechanism.


Increased Scholarly Activity: A Benefit of 4-Year Family Medicine Residencies.
The optimal length of Family Medicine Residency is unknown. As part of the American Board of Family Medicine 4-year Length of Training (LoT) pilot project, Naval Hospital Jacksonville (NHJ) maintained a dual-track 3- and 4-year Family Medicine Residency, graduating seven 4-year residents over consecutive 4 years of the LoT program. One measure of success regarding the impact of 4-year residents on program outcomes is scholarly output during residency. Cumulative scholarly activity points are tracked for all NHJ residents. Cumulative scholarly activity points, points per year per, and raw percentile USMLE/COMLEX scores from academic years 2016-17 to 2019-20 were compared between PGY3 and PGY4 graduates using one-way ANOVA to 95% confidence with post hoc Tukey honestly significant difference pairwise comparison to evaluate pairwise significance between groups where multi-group differences were found. During the 2016-17 through 2019-20 academic years, NHJ had 28 residents complete 3 years of training without interruption (3 Years), 11 residents complete 3 years of training interrupted by general medical officer tours (Resiterns), and 7 residents complete 4 years of training without interruption (4 Years). There were no significant differences in average raw USMLE and COMLEX scores between 3 Year (71%), Resitern (68%), and 4 Year (76%) residents (P = .335). 4-Year residents had significantly more cumulative scholarly points (103) than 3-Year residents (32.6, P < .001) and Resiterns (18.7, P < .001) and also had more cumulative scholarly points per year of residency (27.8) than 3-Year residents (9.8, P < .001) and Resiterns (7.0, P < .001). An observed benefit of a 4-year Family Medicine Residency was a marked increase in scholarly output at this program.


[The effects of diltiazem on neuromuscular transmission].
The effects of intravenous diltiazem (0.5 and 1.0 mg.kg-1) on muscle compound action potential (CAP) elicited from gastrocnemius muscle by sciatic nerve stimulation in cats were investigated. Analysis was performed on the amplitude of CAP, recovery curve (RC) and train-of-four ratio (TOFR). There was no response in any parameter after administering 0.5 mg.kg-1. On the other hand, diltiazem 1.0 mg.kg-1 produced significant depression only in the amplitude of CAP, but no change was observed in both RC and TOFR. It is concluded that approximately ten times the therapeutic dose of diltiazem is necessary to modify the neuromuscular transmission.


Neutrophils require SHP1 to regulate IL-1β production and prevent inflammatory skin disease.
The regulation of neutrophil recruitment, activation, and disposal is pivotal for circumscribed inflammation. SHP1(Y208N/Y208N) mutant mice develop severe cutaneous inflammatory disease that is IL-1R dependent. Genetic reduction in neutrophil numbers and neutrophilic responses to infection is sufficient to prevent the spontaneous initiation of this disease. Neutrophils from SHP1(Y208N/Y208N) mice display increased pro-IL-1β production due to altered responses to MyD88-dependent and MyD88-independent signals. The IL-1R-dependent inflammatory disease in SHP1(Y208N/Y208N) mice develops independently of caspase 1 and proteinase 3 and neutrophil elastase. In response to Fas ligand, a caspase 1-independent inducer of IL-1β production, neutrophils from SHP1(Y208N/Y208N) mice produce elevated levels of IL-1β but display reduced caspase 3 and caspase 7 activation. In neutrophils deficient in SHP1, IL-1β induces high levels of pro-IL-1β suggesting the presence of a paracrine IL-1β loop. These data indicate that the neutrophil- and IL-1-dependent disease in SHP1(Y208N/Y208N) mice is a consequence of loss of negative regulation of TLR and IL-1R signaling.


Closure of a persistent cyclodialysis cleft using the haptics of a normal-sized intraocular lens.
A 50-year-old man suffering from hypotony in the right eye caused by a traumatic cyclodialysis and complicated by a choroidal detachment and cataract was treated in our clinic. After an unsuccessful direct cyclopexy, phacoemulsification was performed and a normal-sized single-piece polymethyl methacrylate posterior chamber intraocular lens (PMMA PCIOL) was inserted into the ciliary sulcus, with the haptic rotated toward the cyclodialysis cleft. Postoperatively, the corrected visual acuity improved to 20/20, and the intraocular pressure returned to normal. Ultrasound biomicroscopy showed the closure of the cleft. Phacoemulsification with a normal-sized PMMA PCIOL inserted into the ciliary sulcus is a safe, effective and technically simple surgical treatment for small cyclodialysis induced hypotony complicated by cataract. Internal compression of the cleft by the haptic of a normal sized IOL along with postoperative inflammation led to scarring and closure of the cleft.


Achromatopsia: the first case report in Thailand.
Achromatopsia, also known as rod mon-ochromacy, is a congenital and nonprogressive ocular disorder characterized by an absence of functional cone photoreceptors in the retina. Affected subjects usually present in infancy with photophobia, nystagmus, low visual acuity and inability to discriminate colors. Fundus examination is normal, but electroretinography demonstrates absent photopic (cone) responses and normal scotopic (rod) responses. Achromatopsia is rare in the general population and inherited as an autosomal recessive trait. The authors report the first case of achromatopsia in Thailand with complete electrophysiological findings and long-term follow-up (8 years).


Lessons learned about slow discontinuous conduction from models of impulse propagation.
Propagation of excitation in the heart involves action potential generation by cardiac cells and its propagation in the multicellular tissue. Action potential conduction is the outcome of complex interactions between cellular processes (membrane ion channels and transporters), electrical communication between cells, and the macroscopic architecture of cardiac tissue. This high level of complexity and non linearity requires computer modeling in order to elucidate mechanisms and explain experimentally observed behaviors. This article reviews studies that we have conducted in 1-dimensional models that contain both, cellular ionic processes and intercellular communication through gap junctions. We have defined and computed a quantitative measure of the robustness of conduction, the safety factor, as a function of gap junction coupling and showed that reduced coupling can support very slow and very safe conduction. Such conduction is highly discontinuous and is supported by the L-type calcium current. We also show that gap junction coupling and the associated load has a strong effect on spatial repolarization gradients in cardiac tissue.


[Schistosomal portal hypertension: influence of the portal blood flow in serum levels of hepatic enzymes].
To evaluate relation between the portal blood flow and the laboratory hepatic screening in patients with schistosomal portal hypertension. Sixty-four patients with schistosomal portal hypertension had studied, being 19 not operated, 23 submitted to esophagogastric devascularization with splenectomy and 22 submitted to distal splenorenal shunt. Evaluated the laboratory hepatic screening through the dosage of albumin, aspartate aminotransferase, alanine aminotransferase, direct bilirubin and indirect bilirubin, alkaline phosphatase, gamma-glutamil transferase and prothrombin time. The portal flow was evaluated for Doppler. The results have been analyzed through linear regression, Pearson correlation coefficient, chi-square and one-way analysis of variance with Tukey's test. It was proven that only gamma-glutamil transferase had significant correlation with the portal flow. In compare of the quartiles, also only gamma-glutamil transferase showed resulted significant, it was evidenced that the fourth quartile, that is bigger portal flow and formed in its majority for patients not operated, also was bigger average of gamma-glutamil transferase and significantly bigger value than first and the third quartiles. 1. The portal blood flow was bigger in patients that the serum level of GGT was bigger; 2. the gamma-glutamil transferase is the variable of the hepatic screening evaluation more representative of the portal flow influence in hepatic functional activity in patients with hepatosplenic schistosomiasis, and 3. probably, the different surgeries through hemodynamics modifications, are beneficial in to diminish the degree of cholestasis or in decrease the microssomal induction.


RECENT ADVANCES IN THE PHYSIOLOGY OF GASTRIC ACID SECRETION.
The classic scheme of gastric acid secretion which divided the digestive period into cephalic, gastric and antral phases has become obsolete in the last 10 years. These "phases" are now seen as concurrently acting mechanisms which depend upon one another to be fully efficient. About half of all gastrin released during a meal is dependent upon vagal stimulation of the antrum. Also, vagotomy desensitizes the acid-secreting parietal cells to the effect of all other types of stimuli.The number of parietal cells (parietal cell mass) varies greatly according to the gastric secretory activity of each individual. It is highest with duodenal ulcer and lowest with gastric ulcer.Parietal cell hyperplasia or atrophy can be induced experimentally, but the factors controlling the size of the parietal cell mass in man have not been studied.A scheme of acid secretion which incorporates recent advances is presented.


Successful Removal of Giant Intrapericardial Paraganglioma via Posterolateral Thoracotomy.
Intrapericardial paraganglioma remains a surgical challenge because of its hypervascular nature and firm adhesion to adjacent mediastinal structures. Here, we describe a 63-year-old female with a giant nonfunctioning intrapericardial paraganglioma tightly adhered to the left atrium. Marginal but complete resection of the tumor was achieved via right posterolateral thoracotomy. At the time of dissection between the tumor and the left atrial wall, we encountered massive hemorrhage leading to cardiac arrest. We were able to repair the wall laceration with minimal time under an optimal operative field, which avoids air embolism. She was discharged without complications and is currently in good health with no recurrence or metastasis for 15 months. Based on our experience, cardiopulmonary bypass should be considered, if surgeons are able to secure suitable sites for arterial and venous cannulations while right posterolateral thoracotomy is employed.


Production of a novel antigen by conjugation of HIV-1 to Brucella abortus: studies of immunogenicity, isotype analysis, T-cell dependency, and syncytia inhibition.
In the present study inactivated human immunodeficiency virus type 1 (HIV-1) was conjugated to Brucella abortus and tested for immunogenicity in normal and anti-L3T4-treated BALB/c mice. HIV-BA was more immunogenic than uncoupled HIV in normal mice, since 6-fold less virus in HIV-BA preparations elicited higher titer responses than HIV-1 alone. Furthermore, the HIV-BA antibody response reached higher levels before the HIV-1 response. Immunoblot analysis showed that most of the HIV-1 antigens were recognized by antibodies induced by either HIV-1 or HIV-BA. Isotype analysis revealed that HIV-1 induced similar levels of IgG1 and IgG2a antibodies, whereas the IgG2a responses to HIV-BA were more pronounced than the IgG1 response. These different IgG subclass patterns suggest that conjugation of HIV-1 to BA changed the immunogenic nature of HIV-1. The requirement for helper T cells was examined by immunizing mice that were depleted of CD4+ T cells by in vivo anti-L3T4 treatment. Under these conditions the IgG responses to HIV-1 were completely eliminated. Although HIV-BA antibody responses were markedly reduced in anti-L3T4-treated mice, anti-HIV-1 antibodies, mainly of the IgG2a isotype, were produced. The antibodies generated by HIV-1 and HIV-BA immunization were also tested for their ability to inhibit syncytia formed by infecting CD4 + CEM cells with gp160 vaccinia. Sera from normal mice, immunized with either HIV-1 or HIV-BA were capable of inhibiting syncytia. In contrast, following anti-L3T4 treatment, only mice immunized with HIV-BA, but not HIV-1, produced antibodies capable of inhibiting syncytia.


(2R,6S)-tert-Butyl 2-(benzhydryl-carbamo-yl)-6-methyl-morpholine-4-carboxyl-ate.
The title compound, C(24)H(30)N(2)O(4), was obtained by the reaction of (2R,6S)-4-(tert-but-oxy-carbon-yl)-6-methyl-morpho-line-2-carb-oxy-lic acid with diphenyl-methanamine in dimethyl-formamide solution. The morpholine ring is in a chair conformation. In the crystal, weak inter-molecular C-H⋯O hydrogen bonds link mol-ecules into chains along the b axis.


Intercropping of kenaf and soybean affects plant growth, antioxidant capacity, and uptake of cadmium and lead in contaminated mining soil.
Intercropping is considered a sustainable agricultural practice that can reduce the environmental impacts on agriculture. Our study investigated the morphology, physiology, and cadmium (Cd) and lead (Pb) uptake of kenaf (Hibiscus cannabinus L.) and soybean (Glycine max L.) under intercropping in mining soil. Results showed that mutual intercropping is conducive to the growth and biomass accumulation of kenaf and soybean, compared to their respective monoculture. Intercropping increased the relative chlorophyll index in kenaf, while that in soybean had no significant effect. Furthermore, intercropping increased the antioxidant enzyme activity of kenaf, while that of soybean reduced or had no significant effect. The content of malondialdehyde (MDA) was decreased in both of the species. Compared to their respective monoculture, Cd content was increased in kenaf leaves and reduced in soybean roots. Moreover, intercropping decreased the Pb content in tissues of both the species, except that Pb content of kenaf roots was increased. At the same time, root, leaf, or stem bioconcentration factors also performed the same trend, and TF was less than 1. These results indicated that intercropping can increase the plant growth and decrease the metal content in plant tissues. Present findings could provide support for future research on kenaf and soybean cultivation in contaminated lands. In addition, the present study strengthens our understanding about the effectiveness of intercropping system on heavy metal-contaminated lands for sustainable agricultural production.


Alternative splicing variant of kallikrein-related peptidase 8 as an independent predictor of unfavorable prognosis in lung cancer.
A relatively unexplored area for biomarker identification is alternative splice variants. We undertook this study to evaluate the usefulness of mRNA isoforms encoded by the KLK8 (kallikrein-related peptidase 8) gene as prognostic markers for lung cancer. Real-time reverse-transcription PCR was used to analyze the mRNAs encoded by KLK8 (particularly 2 mRNA splice variants, KLK8-T3 and KLK8-T4) in 60 non-small-cell lung cancer (NSCLC) tumors and in paired unaffected tissues. The ratios of these mRNAs to those encoded by the KLK5, KLK6, KLK7, KLK10, KLK11, KLK13, and KLK14 genes were also determined and analyzed for correlations with various clinicopathologic variables. KLK8-T3 and KLK8-T4 were the most abundant of the 6 mRNA isoforms identified in lung tissues. The overall expression of the KLK8 gene and the amounts of the KLK8-T3 and KLK8-T4 mRNAs were significantly increased in lung tumor tissue (P < 0.0001). Univariate survival analysis revealed significant relationships of the relative concentrations of mRNA splice variants KLK8 (P = 0.043), KLK8-T3 (P = 0.037), and KLK8-T4 (P = 0.009) with overall survival (OS). Cox multivariate analysis indicated that the amount of KLK8-T4 mRNA was an independent prognostic factor for OS (relative risk = 3.90; P = 0.016) and that high KLK8-T4/KLK7, KLK8-T4/KLK10, and KLK8-T4/KLK11 mRNA ratios in NSCLC indicated increased risk of death. The increase was approximately 5-fold for the KLK8-T4/KLK7 and KLK8-T4/KLK10 ratios (P = 0.006, and P = 0.011, respectively) and 8-fold for the KLK8-T4/KLK11 ratio (P = 0.001). The KLK8-T4 alternative splice variant, alone or in combination, may be a new independent marker of unfavorable prognosis in lung cancer.


Patient education documentation: is it being done?
Patient education is a key component to disease management, convalescence, and overall wellness maintenance. Documentation of patient education provides a means of monitoring the type of patient education performed, the patient response, and additional educational needs.


Child Health Outcome Inequalities in Low and Middle Income Countries.
Prior to the Sustainable Development Goals (SDG) era, considerable progress was made toward the Millennium Development Goals (MDGs) health indicators. Despite these achievements, many countries failed to meet the MDG target levels, between-country inequalities in health outcomes did not improve, and many countries making progress in average indicator levels did so while at the same time seeing increasing within-country inequalities. We build on the existing literature documenting levels and trends in health inequalities by expanding the number of data-points under focus, and we contribute to this literature by analyzing the extent to which inequalities in child health outcomes are related to socioeconomic inequalities, and to aggregate income growth. The objective of this paper is to examine long-run trends in average population levels and within-country inequalities for two child health outcomes-the under-five mortality rate (U5MR) and stunting-in 102 countries across 6 regions. We find that only about a third of countries in our sample managed to both reduce U5MR levels and inequalities, and only a quarter did so for stunting. The fact that inequality in service coverage seems to follow a more favorable trend than inequality in health outcomes suggests that policies aiming to reduce health inequities should not only foster more equitable service coverage but also focus on the social determinants of health. Moreover, there is no strong correlation between changes in health inequalities and income growth, suggesting that income generating development policies alone will typically not suffice to improve health outcomes and reduce health inequalities.


Accuracy of novel diagnostic biomarkers for hepatocellular carcinoma: An update for clinicians (Review).
Hepatocellular carcinoma (HCC) is the most common liver malignancy and a leading cause of cancer-related mortality worldwide. Accurate detection and differential diagnosis of early HCC can significantly improve patient survival. Currently, detection of HCC in clinical practice is performed by diagnostic imaging techniques and determination of serum biomarkers, most notably α-fetoprotein (AFP), fucosylated AFP and des-γ-carboxyprothrombin. However, these methods display limitations in sensitivity and specificity, especially with respect to early stages of HCC. Recently, high-throughput technologies have elucidated many new pathways involved in hepatocarcinogenesis and have led to the discovery of a plethora of novel, non-invasive serum biomarkers. In particular, the combination of AFP with these new candidate molecules has yielded promising results. In this review, we aimed at recapitulating the most recent (2013-2015) developments in HCC biomarker research. We compared promising novel diagnostic serum protein biomarkers, such as annexin A2, the soluble form of the receptor tyrosine kinase Axl and thioredoxin, as well as their combinations with AFP. High diagnostic performance (area under the curve >0.75) as shown by threshold-independent receiver operating characteristic curve analysis was a prerequisite for inclusion in this review. In addition, we discuss the role and potential of microRNAs in HCC diagnosis and associated methodological challenges.


Transition metal-free regioselective direct acylation of quinazolines for the synthesis of acylquinazoline derivatives.
An efficient, mild method for direct regioselective acylation of quinazolines under metal-free conditions was developed with bis(trifluoroacetoxy)iodobenzene and trimethylsilyl azide at ambient temperature. The acylation reaction of quinazolines with aldehydes gave the corresponding acyl quinazolines in ethyl acetate with good to excellent yields and excellent functional group tolerance and site selectivity. In addition, the mechanism of the direct acylation of quinazolinone was investigated through HPLC-HRMS (high pressure liquid chromatography-high resolution mass spectrometry) and EPR (electron paramagnetic resonance) strategies.


Comparative analysis of imaging sensitivity of positron emission mammography and whole-body PET in relation to tumor size.
Positron emission mammography (PEM) consists of a dedicated PET scanner for breast imaging with a higher spatial resolution than whole-body PET (WBPET) scanners. This study compared the imaging sensitivity of PEM with WBPET in relation to tumor size. Fifty-four Japanese women younger than 50 years with histologically confirmed breast lesions were retrospectively enrolled. Positron emission mammography and WBPET were conducted on the same day. Positron emission mammography and WBPET images were blindly evaluated and compared with histopathology. Tumors were classified into 3 groups based on size as follows: group 1, 1 cm or smaller; group 2, 1 to 2 cm; and group 3, larger than 2 cm. The sensitivities of PEM and WBPET were compared in overall subjects and in each size group. In visual analysis, the overall imaging sensitivity was 78.6% (33/42) for PEM and 47.6% (20/42) for WBPET. The overall sensitivity of PEM was significantly higher than that of WBPET (P < 0.001). The differences in sensitivities between PEM and WBPET were larger in smaller tumors: group 1 (66.7% vs 13.3%), group 2 (63.4% vs 36.4%), and group 3 (100.0% vs 87.5%). The sensitivity of PEM was significantly higher than that of WBPET in group 1 (P = 0.008); however, no significant differences were seen in group 2 (P = 0.500) or group 3 (P = 0.250). Overall, the imaging specificity of PEM and WEBPET was 90.6% (60/66) and 93.9% (62/66), respectively. The imaging sensitivity of PEM was higher than that of WBPET in Japanese women younger than 50 years. Positron emission mammography showed significant sensitivity in tumors smaller than 1 cm, which has been a weak point for WBPET.