Apoptosis in shed human corneal cells.
To determine whether shear forces applied to the corneal epithelium by the repeated insertion and removal of a hydrogel contact lens alter the size and number of cells removed and to determine the contribution of apoptosis to this process. METHODS; Human corneal cells were collected from eight healthy subjects by sequential contact lens cytology (20 lens insertions and removals). Collected cells were stained with acridine orange for counting and measurement of cell size. In a separate experiment, collected cells were fixed and stained with TdT-mediated dUTP nick-end labeling (TUNEL) or labeled immediately after collection using annexin V. Hoechst stain and propidium iodide (PI) were used as nuclear counterstains. The proportion of cells labeled with acridine orange, TUNEL, and annexin V was quantified by fluorescence microscopy. RESULTS; The number of cells increased in later collections, and cells were smaller. The mean number of positively stained cells using TUNEL was 57%. Annexin V labeling on unfixed fresh samples showed a mean of 64%, with an increase in later collections. Apoptotic bodies were observed in very few cells. In most cells the nucleus and cytoplasmic membrane were intact. Structures were observed in which nuclei were missing (Hoechst negative) but in which cytoplasm had the size and appearance of whole, nucleated cells. These structures (cell ghosts) increased in number along with the increase in nucleated cells in later collections. The sequential removal of a soft contact lens caused a progressive increase in the number of cells collected from the surface and a progressive decrease in their size. The majority of nucleated cells removed by a contact lens were apoptotic in the sense of being positively labeled by TUNEL and annexin V. Morphologically they differed from classically apoptotic cells, in that cells showed an intact nuclear structure and no discernible apoptotic bodies. They could represent a last stage in a pathway of cell differentiation in which frictional forces induced by the removal of the contact lens activate the apoptotic program and cause the cell to be shed. There is also a pathway in which cells lose their nuclei before leaving the epithelial surface.


Infantile or malignant osteopetrosis: case report of two siblings.
Infantile or Malignant osteopetrosis is a rare congenital disorder of bone resorption. It is caused by failure of osteoclasts to reabsorb immature bone. Severe infantile or malignant osteopetrosis present at birth or develops within the first few months of life. We are reporting here a case of two siblings with malignant osteopetrosis. Prominent clinical features included marked pallor, noisy respiration, progressive blindness and developmental delay.


Calcium channel subtypes for cholinergic and nonadrenergic noncholinergic neurotransmission in isolated guinea pig trachea.
The Ca(2+) channel subtypes in the neurotransmission of isolated guinea pig trachea were elucidated by monitoring the effects of specific Ca(2+) channel blockers on cholinergic contractions and nonadrenergic noncholinergic (NANC) relaxation elicited by electrical field stimulation (EFS). In isolated guinea pig trachea, cholinergic contractile responses to low- and high-frequency EFS were inhibited by the selective N-type calcium channel blocker, ω-conotoxin MVIIA. ω-Agatoxin IVA (a selective P-type blocker), ω-conotoxin MVIIC (a nonselective N-, Q-, and P-type blocker), and nifedipine (a selective L-type blocker) were ineffective, whereas Ni(2+) (a T- and R-type blocker) facilitated cholinergic contractions and produced a late contracture when its concentration exceeded 30 μM. The more the concentration of Ni(2+) increased, the greater the number of incidences and the late contracture areas which occurred. Late contracture did not seem to be due to the effects of acetylcholine, tachykinins, or other polypeptides, but disappeared in the absence of indomethacin. The NANC relaxant responses elicited by the low- and high-frequency EFS were inhibited by ω-conotoxin MVIIA or Ni(2+), but unaffected by ω-Agatoxin IVA, ω-conotoxin MVIIC, and nifedipine. In the absence of indomethacin, Ni(2+) did not alter the ω-conotoxin MVIIA (100 nM)-resistant component of cholinergic contraction, but significantly further inhibited that of NANC relaxation. These results suggest that in isolated guinea pig trachea, cholinergic contraction is regulated by N-type calcium channels which may mask T- and R-type calcium channels and may be co-modulated by both, while NANC relaxation is mainly and independently controlled by N-, T-, and R-type calcium channels.


Comparison between endoluminal ultrasonography and spiral computerized tomography for the preoperative local staging of rectal carcinoma.
The aim of this study is to compare the efficacy of endoluminal ultrasonography (EUS) and spiral computerized tomography (SCT) in preoperative local staging of rectal carcinoma. EUS and SCT were performed prior to surgery in 78 patients with rectal carcinoma. After radical surgery, the preoperative findings were compared with histologic findings on the surgical specimen, and we assessed the values of EUS and SCT in staging the tumor. For T staging, accuracy was 84.6% for EUS, 70.5% for SCT (p < 0.05). For N staging, accuracy was 64.1% for EUS, 61.5% for SCT (p > 0.05). EUS is superior to SCT in judging tumor infiltrate depth, but neither could provide satisfactory assessments of lymph node metastases.


Evaluation of a large cohort of adult patients with Ménière's disease: bedside and clinical history.
The purpose of this study was to assess vestibular findings and clinical history in a large cohort of patients affected by Ménière's disease. We retrospectively analysed 511 adult patients fulfilling criteria for definite unilateral Ménière's disease according to Barany Society. Thorough clinical history, audiometric exam, central nervous system MRI, quantification of serum autoantibodies and complete vestibular function test were performed. Mean age at clinical record was 55.4 years, while age at onset of the first vertigo attack was 47.4 ± 14.3 years. Ménière's disease overlapped with migraine in 43.4% of patients. In 31.7% of cases, positivity was found for at least one autoantibody. Forty-nine patients (9.6%) had family history for Ménière's disease. Bedside examination resulted in 14.7% positivity for video head impulse test, 58.9% for skull vibration-induced nystagmus, 38.7% for the positional test and 23.1% for the post head shaking test. Complete negative examination was reported in 115 cases. Ménière's disease was seen to present a characteristic phenotypic pattern in our cohort, confirming the crucial role of thorough anamnesis and bedside examination in diagnosis.


Volume-induced increase of anion permeability in human lymphocytes.
Peripheral blood mononuclear cells (PBM) readjust their volumes after swelling in hypotonic media. This regulatory volume decrease (RVD) is associated with a loss of cellular K+ and is thought to be promoted by an increased permeability to this ion. In contrast, no change in volume was observed when K+ permeability of PBM in isotonic media was increased to comparable or higher levels using valinomycin. Moreover, valinomycin-induced 86Rb+ loss in K+-free medium was considerably slower than in K+-rich medium. These results suggest that anion conductance limits net salt loss in isotonic media. Direct measurements of relative conductance confirmed that in volume-static cells, anion conductance is lower than that of K+. In volume-regulating cells depolarization occurred presumably as a result of increased anion conductance. Accordingly, the efflux of 36Cl from PBM was markedly increased by hypotonic stress. Since both membrane potential and intracellular 36Cl concentration are reduced in hypotonically swollen cells, the increased efflux is probably due to a change in Cl- permeability. Anions and cations seem to move independently through the volume-induced pathways: the initial rate of 86Rb uptake in swollen cells was not affected by replacement of external Cl- by SO=4; conversely, 36Cl fluxes were unaffected by substitution of K+ by Na+. The data indicate that anion conductance is rate-determining in salt and water loss from PBM. An increase in anion conductance is suggested to be the critical step of RVD of human PBM.


Flavonoids activate pregnane x receptor-mediated CYP3A4 gene expression by inhibiting cyclin-dependent kinases in HepG2 liver carcinoma cells.
The expression of the drug-metabolizing enzyme cytochrome P450 3A4 (CYP3A4) is regulated by the pregnane x receptor (PXR), which is modulated by numerous signaling pathways, including the cyclin-dependent kinase (Cdk) pathway. Flavonoids, commonly consumed by humans as dietary constituents, have been shown to modulate various signaling pathways (e.g., inhibiting Cdks). Flavonoids have also been shown to induce CYPs expression, but the underlying mechanism of action is unknown. Here, we report the mechanism responsible for flavonoid-mediated PXR activation and CYP expression. In a cell-based screen designed to identify compounds that activate PXR-mediated CYP3A4 gene expression in HepG2 human carcinoma cells, we identified several flavonoids, such as luteolin and apigenin, as PXR activators. The flavonoids did not directly bind to PXR, suggesting that an alternative mechanism may be responsible for flavonoid-mediated PXR activation. Consistent with the Cdk5-inhibitory effect of flavonoids, Cdk5 and p35 (a non-cyclin regulatory subunit required to activate Cdk5) were expressed in HepG2. The activation of Cdk5 attenuated PXR-mediated CYP3A4 expression whereas its downregulation enhanced it. The Cdk5-mediated downregulation of CYP3A4 promoter activity was restored by flavonoids, suggesting that flavonoids activate PXR by inactivating Cdk5. In vitro kinase assays showed that Cdk5 directly phosphorylates PXR. The Cdk kinase profiling assay showed that apigenin inhibits multiple Cdks, suggesting that several Cdks may be involved in activation of PXR by flavonoids. Our results for the first time link the stimulatory effect of flavonoids on CYP expression to their inhibitory effect on Cdks, through a PXR-mediated mechanism. These results may have important implications on the pharmacokinetics of drugs co-administered with herbal remedy and herbal-drug interactions.


Insulin management in overweight or obese type 2 diabetes patients: the role of insulin glargine.
Type 2 diabetes mellitus (T2DM) and obesity commonly co-exist. Improved clinical management of T2DM and improved glycaemic control with traditional therapies including insulin usually result in some weight gain - a frequently perceived barrier to the introduction of insulin by both patient and healthcare professionals. Weight gain of 2.5 kg per 1% change in haemoglobin A(1c) (HbA(1c)) is common in many studies. Strategies to minimize weight gain, particularly in obese patients, are essential to help patients better manage their diabetes and improve quality of life. Insulin analogues with lower risk of hypoglycaemia and better within-patient variability compared with human insulin may help facilitate reaching treatment goals. Moreover, weight gain can be minimized by earlier insulinization and the use of basal insulin, such as insulin glargine, instead of premixed insulin. Data specific to the obese patient with T2DM are presented; they are currently limited but do indicate that insulin glargine therapy is associated with improved glycaemic control as well as less weight gain than other insulins, such as premixed insulin and prandial insulin regimens. Retrospective subanalyses of earlier trials and ongoing studies would shed further light on the impact of insulin therapy in obese people with T2DM in addition to determination of optimal therapeutic strategies.


The kinetic mechanism of EcoRI endonuclease.
Steady-state parameters governing cleavage of pBR322 DNA by EcoRI endonuclease are highly sensitive to ionic environment, with K(m) and k(cat) increasing 1,000-fold and 15-fold, respectively, when ionic strength is increased from 0.059 to 0.23 M. By contrast, pre-steady-state analysis has shown that recognition, as well as first and second strand cleavage events that occur once the enzyme has arrived at the EcoRI site, are essentially insensitive to ionic strength, and has demonstrated that the rate-limiting step for endonuclease turnover occurs after double-strand cleavage under all conditions tested. Furthermore, processive cleavage of a pBR322 variant bearing two closely spaced EcoRI sites is governed by the same turnover number as hydrolysis of parental pBR322, which contains only a single EcoRI sequence, ruling out slow release of the enzyme from the cleaved site or a slow conformational change subsequent to double-strand cleavage. We attribute the effects of ionic strength on steady-state parameters to nonspecific endonuclease.DNA interactions, reflecting facilitated diffusion processes, that occur prior to EcoRI sequence recognition and subsequent to DNA cleavage.


[Antiphospholipid antibodies--antiphospholipid syndrome. Cause or consequence of thrombosis].
Antiphospholipid antibodies (APA) are heterogeneous immunoglobulines of G, M or A classes with specificity directed towards anionic phospholipids. The APA are associated with a wide variety of diseases. They have been found to represent risk factor for development of arterial and/or veinous thrombosis. The APA would perturbed the biological activities of anionic phospholipid surface in a manner that could decrease the natural anti-coagulant pathway in order to lead to thrombosis. In the latter hypothesis APA would appear as a secondary response to the exposure of phospholipid "self antigens" by activated or damaged blood vascular cells by known stimuli able to induce thrombosis. In these cases APA would therefore constitute a marker of risk of thrombosis. The question which remains to be solved is if APA possess their own pathogenic potential or if they appear as the result of an up-stream pathological event known to be favourable to the development of thrombosis or if both mechanism are involved.


Neural responses to happy, fearful and angry faces of varying identities in 5- and 7-month-old infants.
The processing of facial emotion is an important social skill that develops throughout infancy and early childhood. Here we investigate the neural underpinnings of the ability to process facial emotion across changes in facial identity in cross-sectional groups of 5- and 7-month-old infants. We simultaneously measured neural metabolic, behavioral, and autonomic responses to happy, fearful, and angry faces of different female models using functional near-infrared spectroscopy (fNIRS), eye-tracking, and heart rate measures. We observed significant neural activation to these facial emotions in a distributed set of frontal and temporal brain regions, and longer looking to the mouth region of angry faces compared to happy and fearful faces. No differences in looking behavior or neural activations were observed between 5- and 7-month-olds, although several exploratory, age-independent associations between neural activations and looking behavior were noted. Overall, these findings suggest more developmental stability than previously thought in responses to emotional facial expressions of varying identities between 5- and 7-months of age.


Preclinical and clinical estimates of the basal apoptotic rate of a cancer predict the amount of apoptosis induced by subsequent proapoptotic stimuli.
We hypothesized that the basal apoptotic rate (BAR) of a cancer would predict sensitivity to subsequent proapoptotic stimuli. To explore this, preclinical and clinical BAR assays were developed measuring cumulative apoptotic events through ELISAs for soluble caspase-cleaved cytokeratin 18 (M30) normalized to either cell number increase or total tumor volume, respectively. The BARs of A549, HCC44, and SW1573 non-small cell lung carcinoma cell lines were measured following different pro/antiapoptotic manipulations. In isogenic wild-type and stable knockdown (KD) series, pretreatment BARs were correlated with response to proapoptotic stimuli and compared with established apoptosis assays. Pretreatment and posttreatment serum was available from stereotactic body radiation therapy patients. Caspase inhibition and p53 KDs reduced the BAR, whereas serum deprivation, XIAP, or Bcl2 KDs increased the BAR. The nontreated BAR rank ordering of the XIAP series recapitulated that with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and caspase-3/7 activity assays, and predicted each line's sensitivity to TRAIL or irradiation. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling, however, underestimated basal apoptosis during increased apoptotic stress, and caspase-3/7 activity detected minimal death in the media. P53 KDs with lower nontreated BARs were less sensitive to TRAIL and cisplatinum than wild-type. Stereotactic body radiation therapy increased serum M30 values, and the pretreatment clinical BAR strongly correlated with fold change in M30 on treatment (r = 0.93). M30-based BAR assays reflect apoptosis accurately and are more amenable to clinical application than existing apoptosis assays. The pretreatment BAR correlates with cell and/or tumor sensitivity to extrinsic and intrinsic apoptotic pathway stimulation. Prospective clinical exploration is warranted.


The evolution of genomic GC content undergoes a rapid reversal within the genus Plasmodium.
The genome of the malarial parasite Plasmodium falciparum is extremely AT rich. This bias toward a low GC content is a characteristic of several, but not all, species within the genus Plasmodium. We compared 4283 orthologous pairs of protein-coding sequences between Plasmodium falciparum and the less AT-biased Plasmodium vivax. Our results indicate that the common ancestor of these two species was also extremely AT rich. This means that, although there was a strong bias toward A+T during the early evolution of the ancestral Plasmodium lineage, there was a subsequent reversal of this trend during the more recent evolution of some species, such as P. vivax. Moreover, we show that not only is the P. vivax genome losing its AT richness, it is actually gaining a very significant degree of GC richness. This example illustrates the potential volatility of nucleotide content during the course of molecular evolution. Such reversible fluxes in nucleotide content within lineages could have important implications for phylogenetic reconstruction based on molecular sequence data.


Formation of atmospheric molecular clusters consisting of sulfuric acid and C8H12O6 tricarboxylic acid.
Using computational methods, we investigate the formation of atmospheric clusters consisting of sulfuric acid (SA) and 3-methyl-1,2,3-butanetricarboxylic acid (MBTCA), identified from α-pinene oxidation. The molecular structure of the clusters is obtained using three different DFT functionals (PW91, M06-2X and ωB97X-D) with the 6-31++G(d,p) basis set and the binding energies are calculated using a high level DLPNO-CCSD(T)/Def2-QZVPP method. The stability of the clusters is evaluated based on the calculated formation free energies. The interaction between MBTCA and sulfuric acid is found to be thermodynamically favourable and clusters consisting of 2-3 MBTCA and 2-3 SA molecules are found to be particularly stable. There is a large stabilization of the cluster when the amount of sulfuric acid-carboxylic acid hydrogen bonded interactions is maximized. The reaction free energies for forming the (MBTCA)2-3(SA)2-3 clusters are found to be similar in magnitude to those of the formation of the sulfuric acid-dimethylamine cluster. Using cluster kinetics calculations we identify that the growth of the clusters is essentially limited by a weak formation of the largest clusters studied, implying that other stabilizing vapours are required for stable cluster formation and growth.


Expanded molecular detection of MPL codon p.W515 and p.S505N mutations in myeloproliferative neoplasms.
Patients negative for the JAK2 p.V617F somatic variant are frequently reflexed to testing for MPL exon 10 variants. Detection of these variants via multiplexed allele-specific PCR followed by fragment analysis has been previously published. The present study builds on this concept by improving the detection of the p.W515A variant, adding a second allele-specific primer to detect the p.W515R variant, and incorporating an improved primer for p.S505N detection. The W515 amplification employs 5'-labeled allele-specific forward primers to detect p.W515K, p.W515L, p.W515R, and p.W515A. The p.S505N amplification includes an allele-specific reverse primer with a tail extension. Fragments were subject to capillary electrophoresis on an ABI 3500 Genetic Analyzer and analyzed using GeneMapper 6.0 (Thermo Fisher Scientific). Thirty MPL-negative and 13 MPL-positive samples previously tested by a reference laboratory were tested with the MPL LDT. Results were 100% concordant. The MPL LDT has a limit of detection of at least 5% VAF for the p.W515 variants and 10% VAF for the p.S505N variant. Current MPL assays are predominantly focused on p.W515L/K and p.S505N mutations. We have engineered an MPL test for detecting p.W515A/L/K/R and p.S505N variants, thereby increasing the diagnostic yield with little additional expense or technician time.


Yoga vs Cognitive Processing Therapy for Military Sexual Trauma-Related Posttraumatic Stress Disorder: A Randomized Clinical Trial.
First-line treatment for posttraumatic stress disorder (PTSD) in the US Department of Veterans Affairs (VA), ie, trauma-focused therapy, while effective, is limited by low treatment initiation, high dropout, and high treatment refraction. To evaluate the effectiveness of Trauma Center Trauma-Sensitive Yoga (TCTSY) vs first-line cognitive processing therapy (CPT) in women veterans with PTSD related to military sexual trauma (MST) and the hypothesis that PTSD outcomes would differ between the interventions. This multisite randomized clinical trial was conducted from December 1, 2015, to April 30, 2022, within 2 VA health care systems located in the southeast and northwest. Women veterans aged 22 to 71 years with MST-related PTSD were enrolled and randomized to TCTSY or CPT. The TCTSY intervention (Hatha-style yoga focusing on interoception and empowerment) consisted of 10 weekly, 60-minute group sessions, and the CPT intervention (cognitive-based therapy targeting modification of negative posttraumatic thoughts) consisted of 12 weekly, 90-minute group sessions. Sociodemographic data were collected via self-report survey. The primary outcome, PTSD symptom severity, was assessed using the Clinician-Administered PTSD Scale for DSM-5 (CAPS-5) and PTSD Checklist for DSM-5 (PCL-5). Assessments were conducted at baseline, midintervention, 2 weeks post intervention, and 3 months post intervention. Of 200 women veterans who consented to participate, the intent-to-treat sample comprised 131 participants (mean [SD] age, 48.2 [11.2] years), with 72 randomized to TCTSY and 59 randomized to CPT. Treatment was completed by 47 participants (65.3%) in the TCTSY group and 27 (45.8%) in the CPT group, a 42.6% higher treatment completion rate in the TCTSY group (P = .03). Both treatment groups improved over time on the CAPS-5 (mean [SD] scores at baseline: 36.73 [8.79] for TCTSY and 35.52 [7.49] for CPT; mean [SD] scores at 3 months: 24.03 [11.55] for TCTSY and 22.15 [13.56]) and the PCL-5 (mean [SD] scores at baseline: 49.62 [12.19] for TCTSY and 48.69 [13.62] for CPT; mean [SD] scores at 3 months: 36.97 [17.74] for TCTSY and 31.76 [12.47]) (P < .001 for time effects). None of the group effects or group-by-time effects were significant. Equivalence analyses of change scores were not significantly different between the TCTSY and CPT groups, and the two one-sided test intervals fell within the equivalence bounds of plus or minus 10 for CAPS-5 for all follow-up time points. In this comparative effectiveness randomized clinical trial, TCTSY was equivalent to CPT in reducing PTSD symptom severity, with both groups improving significantly. The higher treatment completion rate for TCTSY indicates its higher acceptability as an effective and acceptable PTSD treatment for women veterans with PTSD related to MST that could address current VA PTSD treatment limitations. ClinicalTrials.gov Identifier: NCT02640690.


Endoscopic management of exophthalmos.
Endoscopic orbital decompression is a valuable technique for treatment of patients with proptosis, exposure keratopathy, or optic neuropathy from Graves' orbitopathy. Compared with traditional open approaches, the endoscopic technique provides enhanced visualization and avoids the morbidity of incisions within the facial skin, oral cavity, or conjunctiva. This article describes the authors' techniques for performing endoscopic orbital decompression.


Retinal non-perfusion in diabetic retinopathy.
Diabetic retinopathy is a major cause of vision loss worldwide and areas of retinal non-perfusion (RNP) are a key pathologic feature of the vascular component of diabetic retinopathy. While there is a need for a more complete understanding of the natural history of RNP development and progression, overall, increasing RNP has been closely linked with worsening DR severity. Both traditional and novel approaches to quantitative image assessment are being explored to advance our understanding of the vascular, physiologic and functional changes associated with progressive RNP. Retinal ischemia secondary to RNP leads to tissue hypoxia and changes in the expression of a host of signalling molecules. Current anti-vascular endothelial growth factor and steroid pharmaceutical agents appear to be unable to reperuse areas of RNP, but may be able to slow the progressive longitudinal accumulation of RNP with regular retreatments. There remains a tremendous unmet need for pharmacotherapies that can slow RNP progression and ultimately reperfuse areas of the non-perfused retina. Towards this end, novel targets including the semaphorin family are being investigated.


[Irisin--a new mediator of energy homeostasis].
Skeletal muscles as an active hormonal compartment in the response of physical activity secrete substances named myokines capable of modulating metabolic processes. Myokines take part in communication between muscles and other tissues. Irisin (Ir) - a newly discovered adipomyokine - is cleaved and secreted to the circulation from a fibronectin type III domain containing protein 5 (FNDC5). The mechanism of Ir action has not been described precisely, and receptors for the molecule are not defined yet, but it has been proposed to promote browning of white adipose tissue into beige fat cells. To date we have distinguished two types of adipose tissue in mammals - white, which not only functions as a store of energy but also can play a pro-inflammatory role (secreting adipokines), and brown adipose tissue. Brown adipose tissue has a high mitochondrial content and can dissipate chemical energy in the form of heat (nonshivering termogenesis). It plays a natural antiobesity role and protects against obesity-related diseases. The development of beige adipose tissue, which in its structure and function is similar to brown adipose tissue, and the possibility to modify its amount through some external factors, are nowadays among the most important targets of research on fat cell biology.


Determination of alpha- and beta-hydroxycarbonyls and dicarbonyls in snow and rain samples by GC/FID and GC/MS employing benzyl hydroxyl oxime derivatization
A flame ionization detector (FID) combined with capillary gas chromatography (GC/FID) and gas chromatography/ mass spectrometry (GC/MS) has been used to identify multifunctional carbonyls in wet precipitation samples. The carbonyl groups were first derivatized to O-benzylhydroxyloxime (BH oxime) by using O-benzylhydroxylamine. The BH oxime derivatives were then treated with N,O-bis(trimethylsilyl)acetamide for the hydroxyl group to derive their TMS ethers. The BH oxime/TMS derivatives were measured using GC/FID as well as GC/MS on positive EI and CI (isobutane was used as CI gas) modes. Three groups of carbonyl compounds (monoaldehydes, dicarbonyls, hydroxycarbonyls) were identified in the samples by using this method. We have identified, for the first time, a group of alpha- and beta-hydroxycarbonyls, glycolaldehyde, hydroxyacetone, and 4-hydroxy-2-butanone, in wet precipitation samples. Concentrations of hydroxycarbonyls ranged from 0.9 to 53.8 microg/L in the precipitation samples. Their concentration level is similar to that of low molecular weight dicarboxylic acids, which have been reported as major water-soluble organic compounds in rain.


PSEN1ΔE9, APPswe, and APOE4 Confer Disparate Phenotypes in Human iPSC-Derived Microglia.
Here we elucidate the effect of Alzheimer disease (AD)-predisposing genetic backgrounds, APOE4, PSEN1ΔE9, and APPswe, on functionality of human microglia-like cells (iMGLs). We present a physiologically relevant high-yield protocol for producing iMGLs from induced pluripotent stem cells. Differentiation is directed with small molecules through primitive erythromyeloid progenitors to re-create microglial ontogeny from yolk sac. The iMGLs express microglial signature genes and respond to ADP with intracellular Ca2+ release distinguishing them from macrophages. Using 16 iPSC lines from healthy donors, AD patients and isogenic controls, we reveal that the APOE4 genotype has a profound impact on several aspects of microglial functionality, whereas PSEN1ΔE9 and APPswe mutations trigger minor alterations. The APOE4 genotype impairs phagocytosis, migration, and metabolic activity of iMGLs but exacerbates their cytokine secretion. This indicates that APOE4 iMGLs are fundamentally unable to mount normal microglial functionality in AD.


Gold nanoparticle enrichment method for identifying S-nitrosylation and S-glutathionylation sites in proteins.
We present a simple method by which gold nanoparticles (AuNPs) are used to simultaneously isolate and enrich for free or modified thiol-containing peptides, thus facilitating the identification of protein S-modification sites. Here, protein disulfide isomerase (PDI) and dual specificity phosphatase 12 (DUSP12 or hYVH1) were S-nitrosylated or S-glutathionylated, their free thiols differentially alkylated, and subjected to proteolysis. AuNPs were added to the digests, and the AuNP-bound peptides were isolated by centrifugation and released by thiol exchange. These AuNP-bound peptides were analyzed by MALDI-TOF mass spectrometry revealing that AuNPs result in a significant enrichment of free thiol-containing as well as S-nitrosylated, S-glutathionylated, and S-alkylated peptides, leading to the unequivocal assignment of thiols susceptible to modification.


Enrichment and Molecular Analysis of Breast Cancer Disseminated Tumor Cells from Bone Marrow Using Microfiltration.
Molecular characterization of disseminated tumor cells (DTCs) in the bone marrow (BM) of breast cancer (BC) patients has been hindered by their rarity. To enrich for these cells using an antigen-independent methodology, we have evaluated a size-based microfiltration device in combination with several downstream biomarker assays. BM aspirates were collected from healthy volunteers or BC patients. Healthy BM was mixed with a specified number of BC cells to calculate recovery and fold enrichment by microfiltration. Specimens were pre-filtered using a 70 μm mesh sieve and the effluent filtered through CellSieve microfilters. Captured cells were analyzed by immunocytochemistry (ICC), FISH for HER-2/neu gene amplification status, and RNA in situ hybridization (RISH). Cells eluted from the filter were used for RNA isolation and subsequent qRT-PCR analysis for DTC biomarker gene expression. Filtering an average of 14×106 nucleated BM cells yielded approximately 17-21×103 residual BM cells. In the BC cell spiking experiments, an average of 87% (range 84-92%) of tumor cells were recovered with approximately 170- to 400-fold enrichment. Captured BC cells from patients co-stained for cytokeratin and EpCAM, but not CD45 by ICC. RNA yields from 4 ml of patient BM after filtration averaged 135ng per 10 million BM cells filtered with an average RNA Integrity Number (RIN) of 5.3. DTC-associated gene expression was detected by both qRT-PCR and RISH in filtered spiked or BC patient specimens but, not in control filtered normal BM. We have tested a microfiltration technique for enrichment of BM DTCs. DTC capture efficiency was shown to range from 84.3% to 92.1% with up to 400-fold enrichment using model BC cell lines. In patients, recovered DTCs can be identified and distinguished from normal BM cells using multiple antibody-, DNA-, and RNA-based biomarker assays.


Emergence of Resistance to Quinolones and β-Lactam Antibiotics in Enteroaggregative and Enterotoxigenic Escherichia coli Causing Traveler's Diarrhea.
The objective of this study was to assess the antimicrobial resistance of enteroaggregative Escherichia coli (EAEC) and enterotoxigenic E. coli (ETEC) strains causing traveler's diarrhea (TD) and to investigate the molecular characterization of antimicrobial resistance genes to third-generation cephalosporins, cephamycins, and quinolones. Overall, 39 EAEC and 43 ETEC clinical isolates were studied. The susceptibilities of EAEC and ETEC against ampicillin, amoxicillin-clavulanic acid, cefotaxime, imipenem, chloramphenicol, tetracycline, co-trimoxazole, nalidixic acid, ciprofloxacin, azithromycin, and rifaximin were determined. All genes encoding resistance determinants were detected by PCR or PCR plus DNA sequencing. The epidemiology of selected EAEC and ETEC strains was studied using multilocus sequence typing (MLST). The resistance to quinolones of EAEC and ETEC strains causing TD has significantly increased over the last decades, and high percentages have been found especially in patients traveling to India and sub-Saharan Africa. Sequence type 38 (ST38) and ST131, carrying the blaCTX-M-15 and blaCTX-M-27 genes, respectively, are highly prevalent among extended-spectrum β-lactamase (ESBL)-producing EAEC and ETEC strains. The cephamycinase ACT-20 is described in the present study for the first time in EAEC and ETEC strains causing TD in patients who had traveled to Central America. The percentages of resistance to azithromycin in EAEC and ETEC isolates from patients to Southeast Asia/India and Africa are above 25%. Meanwhile, rifaximin is still active against EAEC and ETEC, with the prevalence of resistant strains not being high. In conclusion, fluoroquinolones should no longer be considered the drugs of choice for the prevention or treatment in TD for travelers traveling to India and Africa. Azithromycin and rifaximin are still a good alternative to treat TD caused by EAEC or ETEC.


Normal conditioned inhibition and extinction of freezing and fear-potentiated startle following electrolytic lesions of medical prefrontal cortex in rats.
The authors investigated the role of medial prefrontal cortex (mPFC) in the inhibition of conditioned fear in rats using both Pavlovian extinction and conditioned inhibition paradigms. In Experiment 1, lesions of ventral mPFC did not interfere with conditioned inhibition of the fear-potentiated startle response. In Experiment 2, lesions made after acquisition of fear conditioning did not retard extinction of fear to a visual conditioned stimulus (CS) and did not impair "reinstatement" of fear after unsignaled presentations of the unconditioned stimulus. In Experiment 3, lesions made before fear conditioning did not retard extinction of fear-potentiated startle or freezing to an auditory CS. In both Experiments 2 and 3, extinction of fear to contextual cues was also unaffected by the lesions. These results indicate that ventral mPFC is not essential for the inhibition of fear under a variety of circumstances.


Organic Material Distribution in Mars-Analog Volcanic Rocks, as Determined with Ultraviolet Laser-Induced Fluorescence Spectroscopy.
Understanding the distribution of trace organic material in a rocky environment is a key to constraining the material requirements for sustaining microbial life. We used an ultraviolet laser-induced fluorescence (LIF) spectroscopy instrument to characterize the distribution of organic biosignatures in basalts collected from two Mars-analog environments. We correlated the fluorescence results with alteration-related sample properties. These samples exhibit a range of alteration conditions found in the volcanic environments of Hawai'i Volcanoes National Park, Hawai'i (HI), and Craters of the Moon National Monument, Idaho (ID), including fumarolic systems. LIF mapping of the sample surfaces and interiors showed a heterogeneous distribution of areas of highly fluorescent material (point[s]-of-interest [POIs])-with fluorescence characteristics indicative of organic material. Results suggest that POIs are associated with secondary alteration mineral deposits in the rock's vesicles, including zeolites and calcite. Scanning electron microscopy with electron-dispersive X-ray spectroscopy was used to characterize the mineralogy present at POIs and support the evidence of carbon-bearing material. Overall, samples collected proximate to active or relict meteoric fumaroles from Hawai'i were shown to contain evidence for organic deposits. This suggests that these minerals are measurable spectroscopic targets that may be used to inform sample-site selection for astrobiology research.


Learning to cope with stress modulates anterior cingulate cortex stargazin expression in monkeys and mice.
Intermittent mildly stressful situations provide opportunities to learn, practice, and improve coping with gains in subsequent emotion regulation. Here we investigate the effects of learning to cope with stress on anterior cingulate cortex gene expression in monkeys and mice. Anterior cingulate cortex is involved in learning, memory, cognitive control, and emotion regulation. Monkeys and mice were randomized to either stress coping or no-stress treatment conditions. Profiles of gene expression were acquired with HumanHT-12v4.0 Expression BeadChip arrays adapted for monkeys. Three genes identified in monkeys by arrays were then assessed in mice by quantitative real-time polymerase chain reaction. Expression of a key gene (PEMT) involved in acetylcholine biosynthesis was increased in monkeys by coping but this result was not verified in mice. Another gene (SPRY2) that encodes a negative regulator of neurotrophic factor signaling was decreased in monkeys by coping but this result was only partly verified in mice. The CACNG2 gene that encodes stargazin (also called TARP gamma-2) was increased by coping in monkeys as well as mice randomized to coping with or without subsequent behavioral tests of emotionality. As evidence of coping effects distinct from repeated stress exposures per se, increased stargazin expression induced by coping correlated with diminished emotionality in mice. Stargazin modulates glutamate receptor signaling and plays a role in synaptic plasticity. Molecular mechanisms of synaptic plasticity that mediate learning and memory in the context of coping with stress may provide novel targets for new treatments of disorders in human mental health.


Characterization of dysgeusia and xerostomia in patients with multiple myeloma treated with the T-cell redirecting GPRC5D bispecific antibody talquetamab.
In recent years, various immunotherapies have improved the survival of patients with multiple myeloma (MM). However, there remains an unmet need for novel agents. Talquetamab is the first-in-class GPRC5D-targeting T-cell redirecting bispecific antibody, which has substantial activity in advanced MM. Rapidly after the start of talquetamab treatment, patients reported taste changes (dysgeusia; 60% of patients), and a feeling of dry mouth (xerostomia; 30-57% of patients), which may be related to expression of the target antigen in healthy tissues, such as taste buds. Here, we aimed at better characterizing these oral toxicities. We measured salivary flow and the ability to taste (objectively and patient-reported), assessed the feeling of dry mouth, and evaluated quality of life before and 8 weeks after the start of talquetamab therapy in eight heavily pretreated MM patients. Talquetamab treatment led to the rapid and significant decrease in objectively measured taste scores (total score 8.8 ± 2.0 vs 4.9 ± 2.5). All patients reported moderate to severe taste changes. Moreover, patients experienced severe xerostomia after the initiation of talquetamab treatment, in the absence of changes in unstimulated and stimulated salivary flow. Because of these oral toxicities a significant impairment in global health status/(oral health related) quality of life was reported. Studying taste changes in patients treated with talquetamab following up on the described leads provides a new and unique opportunity to further unravel the pathophysiology of taste changes after cancer treatment.


The Effect of Various Fluoride Products on Dentine Lesions during pH-Cycling.
This study compared the effect of topically applied fluoride products on dentine lesions in an in vitro experiment. Demineralized bovine dentine specimens were treated once with either SDF solution (35,400 ppm F), NaF varnish (22,600 ppm F), TiF4 solution (9,200 ppm F), SnF2 gel (1,000 ppm F), no treatment (control), or preserved as baseline lesions. After the application and subsequent removal of the fluoride products, the specimens were subjected to pH-cycling. Calcium loss and uptake in the de- and remineralization buffers were assessed daily. Fluoride release into the buffers was analyzed on days 1, 2, 3, 5, 8, and 13. After the pH-cycling period, mineral distribution throughout the lesion depth was analyzed using transversal microradiography (TMR). X-ray energy-dispersive spectroscopy (EDS) examined the deposition of silver, titanium, and tin after application of SDF, TiF4, and SnF2, respectively. Overall, calcium loss and uptake analysis in the de- and remineralization buffers revealed that the SDF product was the most effective in inhibiting lesion progression, followed by the TiF4, NaF, and SnF2 products. Fluoride analysis disclosed a steep reduction of the amount of fluoride released into de- and remineralization buffers with time. The fluoride effects on de- and remineralization continued beyond the days that fluoride was released into the buffers. TMR analysis showed significant remineralization in the outer zone of the dentine lesions for all fluoride products, with SDF giving hypermineralization in this zone. In the inner zone, lesions developed in all fluoride groups, with the smallest in the SDF group. EDS showed silver and titanium deposition in depth up to 85 μm and 8 μm, respectively, while no tin deposition was observed. The silver in the dentine lesions did not contribute significantly to the density of the TMR profiles in the SDF group. In conclusion, all topical fluoride products protected the dentine lesions against lesion progression, but at different degrees. SDF showed a superior effect in protection against further demineralization and enhancement of remineralization. This was probably attributed to its fluoride concentration that was the highest among the fluoride products.


Incidence of presentation of common functional gastrointestinal disorders in children from birth to 5 years: a cohort study.
Gastroesophageal reflux disease (GERD), abdominal pain of unknown origin, and constipation are thought to be causes for frequent medical visits during childhood. The aim of this study was to estimate the incidences, repeat presentation, clinical symptoms, and sociodemographic risk factors in children who medically presented for GERD, abdominal pain of unknown origin, and constipation from birth to 5 years. This was a population-based, retrospective birth cohort study of all children born to mothers residing in Rochester, Minnesota who remained in the area until at least age 5 (n = 5718). The medical records of all individuals were searched for relevant diagnostic billing codes for GERD, abdominal pain of unknown origin, and constipation, without another underlying diagnosis, and manually reviewed. Parental sociodemographic factors collected from birth certificate records on patients and matched controls were compared. The incidence for childhood (age, <5 y) presentation of GERD, abdominal pain of unknown origin, and constipation was .9/1000 person-years, 4.5/1000 person-years, and 6.8/1000 person-years, respectively; there were no significant differences between boys and girls. Three or more medical visits by age 5 occurred in 11%, 19%, and 24% of children who were seen for abdominal pain of unknown origin, constipation, and GERD, respectively. Single parentage, maternal age (<18 y), and maternal education (<high school) were not associated with abdominal pain or constipation presentation. The incidences of presentation for GERD, abdominal pain of unknown origin, and constipation are among the highest for pediatric disorders, and a cause for repeated medical consultations.


[Distribution of anemia among preschool children from the semi-arid region of Bahia]
The objective of this survey was to determine the prevalence of anemia and potential determinants thereof in 754 children from urban areas of seven small towns in the semi-arid region of Bahia. Hemoglobin was measured in 745 children 1 to 72 months of age. For the 6-to-72 month-old group, hemoglobin < 11.0 g/dl was considered anemia (following WHO recommendations) whereas <9.5 g/dl was considered severe anemia. The same cutoffs were used for children under 6 months, which are the same ones used in clinical-hematology. A mean hemoglobin of 12.1 g/dl was found, distributed differently according to age groups (p=0.001). The study found prevalences of 22.2 % for anemia and 5.8% for severe anemia, respectively. Occurrence of anemia varied significantly with age (p=0.001); the highest prevalence was 50.0% in children 12 to 23 months of age, followed by 29.9% in children below 12 months. The association of anemia with mother's education (controlled for age) and per capita family income was not statistically significant.


Brief therapy for children's mental health problems: outcomes in a rural setting.
In rural areas where specialist services are not readily accessible, there are practical advantages to providing short courses of therapy for children's mental health problems. Fifty-five parents from a rural area in South Australia completed questionnaires before and 3 months after receiving brief, family-oriented therapy from a child and adolescent mental health service. Clinically, these families were assessed as being suitable for brief therapy because the children had moderately severe emotional and behavioural problems and the families were not severely dysfunctional. The screening process sought to exclude more serious and complex presentations so those families were not included in the current study. Questionnaires measuring the severity of children's mental health problems (Child Behavior Checklist) and the degree of family dysfunction (Family Assessment Device general functioning subscale) suggested that, on average, the sample met these entry criteria. The follow-up evaluation after brief therapy found significant overall improvement in the Child Behavior Checklist scores, however some children continued to show substantial difficulties. Parents' satisfaction with therapy was related to these scores after treatment. The results suggest that brief therapy was appropriate for most of the families and that routine follow-up could help determine those who might benefit from further assistance.


Benefits of Cardiac Rehabilitation on Cardiovascular Outcomes in Patients With Diabetes Mellitus After Percutaneous Coronary Intervention.
Participation in cardiac rehabilitation (CR) is an essential component of care for patients with coronary artery disease. However, little is known about its benefit on cardiovascular outcomes in patients with diabetes mellitus (DM) who have undergone percutaneous coronary intervention. The aim of our study was to evaluate the impact of CR in this high-risk group of patients. We performed a retrospective analysis of all patients with DM who underwent percutaneous coronary intervention in Olmsted County (Minnesota) between 1994 and 2010, assessing the impact of CR participation on clinical outcomes. CR participation was significantly lower in patients with DM (38%, 263/700) compared with those who did not have DM (45%, 1071/2379; P=0.004). Using propensity score adjustment, we found that in patients with DM, CR participation was associated with significantly reduced all-cause mortality (hazard ratio, 0.56; 95% confidence interval, 0.39-0.80; P=0.002) and composite end point of mortality, myocardial infarction, or revascularization (hazard ratio, 0.77; 95% confidence interval, 0.60-0.98; P=0.037), during a median follow-up of 8.1 years. In patients without DM, CR participation was associated with a significant reduction in all-cause mortality (hazard ratio, 0.67; 95% confidence interval, 0.55-0.82; P<0.001) and cardiac mortality (hazard ratio, 0.67; 95% confidence interval, 0.47-0.95; P=0.024). CR participation after percutaneous coronary intervention is associated with lower all-cause mortality rates in patients with DM, to a similar degree as for those without DM. However, CR participation was lower in patients with DM, suggesting the need to identify and correct the barriers to CR participation for this higher-risk group of patients.


Serial analysis of gene expression in the silkworm, Bombyx mori.
The silkworm Bombyx mori is one of the most economically important insects and serves as a model for Lepidoptera insects. We used serial analysis of gene expression (SAGE) to derive profiles of expressed genes during the developmental life cycle of the silkworm and to create a reference for understanding silkworm metamorphosis. We generated four SAGE libraries, one from each of the four developmental stages of the silkworm. In total we obtained 257,964 SAGE tags, of which 39,485 were unique tags. Sorted by copy number, 14.1% of the unique tags were detected at a median to high level (five or more copies), 24.2% at lower levels (two to four copies), and 61.7% as single copies. Using a basic local alignment search tool on the EST database, 35% of the tags matched known silkworm expressed sequence tags. SAGE demonstrated that a number of the genes were up- or down-regulated during the four developmental phases of the egg, larva, pupa, and adult. Furthermore, we found that the generation of longer cDNA fragments from SAGE tags constituted the most efficient method of gene identification, which facilitated the analysis of a large number of unknown genes.


Dynamic time-varying analysis of heart rate and blood pressure variability in cats exposed to short-term chronic intermittent hypoxia.
Chronic intermittent hypoxia (CIH) contributes to the development of hypertension in patients with obstructive sleep apnea and animal models. However, the early cardiovascular changes that precede CIH-induced hypertension are not completely understood. Nevertheless, it has been proposed that one of the possible contributing mechanisms to CIH-induced hypertension is a potentiation of carotid body (CB) hypoxic chemoreflexes. Therefore, we studied the dynamic responses of heart rate, blood pressure, and their variabilities during acute exposure to different levels of hypoxia after CIH short-term preconditioning (4 days) in cats. In addition, we measured baroreflex sensitivity (BRS) on the control of heart rate by noninvasive techniques. To assess the relationships among these indexes and CB chemoreflexes, we also recorded CB chemosensory discharges. Our data show that short-term CIH reduced BRS, potentiated the increase in heart rate induced by acute hypoxia, and was associated with a dynamic shift of heart rate variability (HRV) spectral indexes toward the low-frequency band. In addition, we found a striking linear correlation (r = 0.97) between the low-to-high frequency ratio of HRV and baseline. CB chemosensory discharges in the CIH-treated cats. Thus, our results suggest that cyclic hypoxic stimulation of the CB by short-term CIH induces subtle but clear selective alterations of HRV and BRS in normotensive cats.


Phosphorelays provide tunable signal processing capabilities for the cell.
Achieving a complete understanding of cellular signal transduction requires deciphering the relation between structural and biochemical features of a signaling system and the shape of the signal-response relationship it embeds. Using explicit analytical expressions and numerical simulations, we present here this relation for four-layered phosphorelays, which are signaling systems that are ubiquitous in prokaryotes and also found in lower eukaryotes and plants. We derive an analytical expression that relates the shape of the signal-response relationship in a relay to the kinetic rates of forward, reverse phosphorylation and hydrolysis reactions. This reveals a set of mathematical conditions which, when satisfied, dictate the shape of the signal-response relationship. We find that a specific topology also observed in nature can satisfy these conditions in such a way to allow plasticity among hyperbolic and sigmoidal signal-response relationships. Particularly, the shape of the signal-response relationship of this relay topology can be tuned by altering kinetic rates and total protein levels at different parts of the relay. These findings provide an important step towards predicting response dynamics of phosphorelays, and the nature of subsequent physiological responses that they mediate, solely from topological features and few composite measurements; measuring the ratio of reverse and forward phosphorylation rate constants could be sufficient to determine the shape of the signal-response relationship the relay exhibits. Furthermore, they highlight the potential ways in which selective pressures on signal processing could have played a role in the evolution of the observed structural and biochemical characteristic in phosphorelays.


Revisiting multi-view learning: A perspective of implicitly heterogeneous Graph Convolutional Network.
Graph Convolutional Network (GCN) has become a hotspot in graph-based machine learning due to its powerful graph processing capability. Most of the existing GCN-based approaches are designed for single-view data. In numerous practical scenarios, data is expressed through multiple views, rather than a single view. The ability of GCN to model homogeneous graphs is indisputable, while it is insufficient in facing the heterophily property of multi-view data. In this paper, we revisit multi-view learning to propose an implicit heterogeneous graph convolutional network that efficiently captures the heterogeneity of multi-view data while exploiting the powerful feature aggregation capability of GCN. We automatically assign optimal importance to each view when constructing the meta-path graph. High-order cross-view meta-paths are explored based on the obtained graph, and a series of graph matrices are generated. Combining graph matrices with learnable global feature representation to obtain heterogeneous graph embeddings at various levels. Finally, in order to effectively utilize both local and global information, we introduce a graph-level attention mechanism at the meta-path level that allocates private information to each node individually. Extensive experimental results convincingly support the superior performance of the proposed method compared to other state-of-the-art approaches.


The challenge of managed care in rural West Virginia.
West Virginia faces a particularly difficult challenge in organizing its health care system for managed care. The rural nature of the state, the shortage of primary care practitioners, few large employers, and the predominant role played by federal and state health care purchasers have hindered managed care development in the state. The crisis in the cost of health care and discussions of health care reform have now created interest in developing prepaid managed care by both payers and providers. In the last three years, several initiatives have paved the way for a more organized approach to the delivery of health care. This article describes the current health care environment in West Virginia and the efforts underway to prepare for system change.


T-cell chronic lymphocytic leukaemia with pulmonary involvement and relapsing BOOP.
We report on a case of T-cell chronic lymphocytic leukaemia involving the lung, with clinical, radiological and histological evidence of relapsing bronchiolitis obliterans-organizing pneumonia in a 70-yr-old female. Pulmonary disease was the major clinical manifestation of this chronic lymphocytic leukaemia. The first two episodes of the patient's pulmonary disorder resolved without treatment, and the third episode was treated with cytotoxic agents as part of the leukaemia treatment regimen. Two additional episodes of the pulmonary disorder occurred; both responded to prednisone.


Dose-dense doxorubicin and cyclophosphamide followed by weekly paclitaxel with trastuzumab and lapatinib in HER2/neu-overexpressed/amplified breast cancer is not feasible because of excessive diarrhea.
Dose-dense doxorubicin and cyclophosphamide (AC) followed by paclitaxel and trastuzumab (PT) is feasible. Lapatinib is effective in the treatment of human epidermal growth factor receptor 2 (HER2) -positive metastatic breast cancer. We conducted a pilot study of dose-dense AC followed by PT plus lapatinib (PTL) followed by trastuzumab plus lapatinib (TL). Patients with stages I to III, HER2-positive breast cancer and left ventricular ejection fraction (LVEF) of > or = 50% were enrolled. Treatment consisted of AC (60 mg/m(2) and 600 mg/m(2)) for 4 cycles every 2 weeks (with pegfilgrastim 6 mg on day 2) followed by paclitaxel (80 mg/m(2)) for 12 doses weekly plus trastuzumab and lapatinib. Trastuzumab (4 mg/kg loading dose, then 2 mg/kg weekly during paclitaxel then 6 mg/kg every 3 weeks after paclitaxel) and lapatinib (1,000 mg daily) were given for 1 year. The primary end points were feasibility defined as > or = 80% patients completing the PTL phase without a dose delay/reduction and a cardiac event rate of < or = 4%. From March 2007 to April 2008, we enrolled 95 patients. Median age was 46 years (range, 28 to 73 years). At a median follow-up of 22 months, 92 were evaluable. Of the 92 patients, 41 patients (45%) withdrew for PTL-specific toxicities. Overall, 40 (43%) of 92 patients had lapatinib dose reductions, and 27 (29%) of 92 patients had grade 3 diarrhea. Three patients (3%) had congestive heart failure; three patients dropped out because of significant asymptomatic LVEF decline during PTL followed by TL. Dose-dense AC followed by PTL and then followed by TL was not feasible because of a high rate of lapatinib dose reduction, mostly caused by unacceptable grade 3 diarrhea. Lapatinib (1,000 mg/d) was not feasible combined with weekly PT.


Induced eosinophilia and proliferation in Angiostrongylus cantonensis-infected mouse brain are associated with the induction of JAK/STAT1, IAP/NF-kappaB and MEKK1/JNK signals.
Eosinophilic meningitis or meningoencephalitis caused by Angiostrongylus cantonensis is endemic to the Pacific area of Asia, especially Taiwan, Thailand, and Japan. Although eosinophilia is an important clinical manifestation of A. cantonensis infection, the role of eosinophils in the progress of the infection remains to be elucidated. In this experiment, we show that A. cantonensis-induced eosinophilia and inflammation might lead to the induction of IAP/NF-kappaB, JAK/STAT1 and MEKK1/JNK signals. The phosphorylation levels of JAK and JNK, STAT1, IAP, NF-kappaB and MEKK1 protein products were significantly increased after 12 days or 15 days of A. cantonensis infection. However, no significant differences in MAPKs such as Raf, MEK-1, ERK1/2 and p38 expression were found between control and infected mice. The activation potency of JAK/STAT1, IAP/NF-kappaB and MEKK1/JNK started increasing on day 3, with significant induction on day 12 or day 15 after A. cantonensis infection. Consistent results were noted in the pathological observations, including eosinophilia, leukocyte infiltration, granulomatous reactions, and time responses in the brain tissues of infected mice. These data suggest that the development of brain injury by eosinophilia of A. cantonensis infection is associated with activation of JAK/STAT1 signals by cytokines, and/or activation of MEKK1/JNK by oxidant stress, and/or activation of NF-kappaB by increasing IAP expression.


Can Dynamic Ultrasonography of the Hip Reliably Assess Anterior Femoral Head Translation?
Hip microinstability has gained attention recently as a potential cause of hip pain. Currently there is a lack of evidence-based objective diagnostic criteria surrounding this diagnosis. Previous studies have shown translation of the femoral head during extreme hip positions. However, reliable assessment of femoral head translation is lacking. (1) How precise is musculoskeletal ultrasound for measuring anterior femoral head translation during the hip anterior apprehension test? (2) What is the intra- and interrater reliability of dynamic ultrasonography in assessing anterior femoral head translation? We recruited 10 study participants (20 hips) between the ages of 22 and 50 years with no history of hip pain or functional limitations. Test-retest methodology was used. Seven females and three males were enrolled. The mean age of study participants was 27 years (SD 8.7 years); mean body mass index was 22.6 kg/m (SD 2.2 kg/m). All study participants underwent dynamic hip ultrasonography by three different physicians 1 week apart. Each hip was visualized in two neutral positions (neutral and neutral with the contralateral hip flexed [NF]) and two dynamic positions, which sought to replicate the apprehension test, although notably study participants had no known hip pathology and therefore no apprehension. The first maintained the hip in extension and external rotation off to the side of the examination table (EER1), and the second held the hip off of the bottom of the examination table (EER2). One hundred twenty ultrasound scans (480 images) were performed. Mean and SD were calculated using absolute values of the difference in ultrasound measurements (mm) between positions NF and EER1 and NF and EER2 calculated for each physician as well as an average of all three physicians. Intraclass correlation coefficient (ICC) analysis was used to examine intra- and interrater reliability. The mean absolute difference for NF and EER1 was 0.84 mm (SD 0.93 mm) and for NF and EER2 0.62 mm (SD 0.40 mm) on Study Day 1. Similarly, on Study Day 2, the mean absolute difference for NF and EER1 position was 0.90 mm (SD 0.74 mm) and for NF and EER2 1.03 mm (SD 1.18 mm). Cumulative values of ICC analysis indicated excellent intrarater reliability in all four positions: neutral 0.794 (95% confidence interval [CI], 0.494-0.918), NF 0.927 (95% CI, 0.814-0.971), EER1 0.929 (95% CI, 0.825-0.972), and EER2 0.945 (95% CI, 0.864-0.978). Similarly, interrater ICC analysis cumulative values were excellent for NF, EER1, and EER2 and fair to good for the neutral position: neutral 0.725 (95% CI, 0.526-0.846), NF 0.846 (95% CI, 0.741-0.913), EER1 0.812 (95% CI, 0.674-0.895), and EER2 0.794 (95% CI, 0.652-0.884). This study offers the first ultrasound protocol of which we are aware for measuring anterior femoral head translation. Hip dynamic ultrasound may assist in providing precise objective clinical-based diagnostic evidence when evaluating complex hip pain and suspected microinstability. Musculoskeletal ultrasound is a reliable office-based method of measuring anterior femoral head translation that can be utilized by physicians with varying experience levels. Future studies are needed to investigate ultrasound anterior femoral head translation taking into account sex, prior hip surgery, hip osseous morphology, and ligamentous laxity. Level III, diagnostic study.


[Experimental antioxidant pharmacotherapy of liver injuries].
It has been shown in rats with experimental hepatitis that sodium selenite and vegetable polyphenol complexes of Malus Pallasiana Juz., Schabiosa comosa F ex. R. et Sch., Odontites serotina Dum., Gentiana barbata Froel, have antioxidant action. Experiments on rabbits and rats have demonstrated the pathogenetic importance of activation of free-radical lipid oxidation during ischemic and toxic injuries to the liver and biliferous ducts.


Covalent attachment of multilayers (CAM): a platform for pH switchable antimicrobial and anticoagulant polymeric surfaces.
These studies show covalent attachment of multilayers (CAM) to chemically alter surfaces to achieve pH switchable antimicrobial and anticoagulant properties. Polyethylene (PE), poly(tetrafluoroethylene) (PTFE), and silicon (Si) surfaces were functionalized by tethering pH-responsive "switching" polyelectrolytes consisting of poly(2-vinyl pyridine) (P2VP) and poly(acrylic acid) (PAA) terminated with NH2 and COOH groups, respectively. At pH < 2.3, the P2VP segments are protonated and expended, but at pH > 5.5, they collapse while the PAA segments are expanded. The presence of terminal NH2 or COOH moieties on P2VP and PAA, respectively, facilitated the opportunity for covalently bonding ampicillin (AMP) and heparin (HEP) to both polyelectrolyte chains. Such surfaces, when exposed to S. aureus, inhibit the growth of microbial films (AMP) as well as anticoagulant properties (HEP). Comparison of "dynamic" pH dependent surfaces developed in these studies with "static" surfaces terminated with (AMP) entities shows significant enhancement in the longevity of surface activity against microbial film formation.


Need For A National Cancer Registry In Pakistan: Challenges And Way Forward.
The incidence of cancer is rising globally at a remarkable rate. According to the global cancer Observatory, the total number of cases in Pakistan were 178,388 in 2020; and these figures were provided by the Punjab Cancer Registry which represents the population of Lahore and some parts of central Punjab. Considering the overall 220 million population, these figures are severe underestimation of the true cancer burden in Pakistan. Unfortunately, several attempts at establishing a national cancer registry in the country have failed in the past. Resource limitations, including funds, manpower and infrastructure, and the lack of interest on the part of the governments are behind the failure of the establishment of a national cancer database. There is a dire need for establishing a national cancer registry to understand the actual burden of disease, and to implement a national action plan for cancer prevention, and management strategies to control the growing epidemic.


Individual differences predictive of drinking to manage anxiety among non-problem drinkers with panic disorder.
Alcohol problems co-occur with anxiety disorders at a rate that far exceeds chance ("comorbidity"). One view suggests that risk for developing a comorbid alcohol use disorder is increased when alcohol is used routinely to cope with anxiety symptoms ("self-medication"). Indicating that this view is overly broad, however, the literature suggests that only a subgroup of anxiety-disordered individuals tend to drink to manage their symptoms. Therefore, we set out to identify psychological characteristics that might mark those for whom drinking to cope with anxiety is most likely. Our survey of the literature identified several possibilities, including anxiety-related personality traits (anxiety sensitivity, self-consciousness and Trait Anxiety); higher-order personality dimensions (Harm Avoidance, Reward Dependence, and Novelty Seeking); and, finally, alcohol outcome expectancies (specifically, those related to tension-reducing effects from alcohol). In a sample of nonproblem drinkers with panic disorder, we regressed predictor variables on several alcohol use measures, including drinking aimed at the control of anxiety symptoms ("self-medication"). Although each variable related positively to a self-medicating style of drinking, expectancies for tension reduction from alcohol use accounted for about twice as much variance as did the other predictors. With simultaneous entry in a step-wise regression analysis, only tension-reduction alcohol outcome expectancies and the Harm Avoidance personality dimension were retained as significant predictors. Findings suggest that development of a self-medicating style of drinking among individuals with panic disorder is predicated, in part, on specific psychological characteristics of the individual. Alcohol outcome expectancies emerged as the single most important predictor of drinking behavior in this anxiety-disordered sample.


[Electrophysiological diagnosis of the caudal cranial nerves].
The lower cranial nerves (CN. IX-XII) are responsible for the nerve supply of the pharyngolaryngeal region. Loss of innervation is manifested by dysphagia, aspiration, altered speech production, dyspnea and/or stridor, and visible loss of movement. In general electrophysiological testing should be applied in every kind of paresis. Beside standard equipment for electromyography limited special equipment is necessary. Examination under local anaesthesia is possible and can serve as a basis for therapeutic consequences. In this article general aspects as seen from an oto-rhino-laryngological standpoint are presented.


Experimental surface or volume deposition of radionuclides in post-mortem specimens of trabecular bone.
Techniques are described that have been developed to contaminate post-mortem specimens of human trabecular bone uniformly and artificially with three different radionuclides, 14C, 45Ca and 32P, so as to produce either a uniform volume deposition of radionuclide through the bone matrix or an 'infinitely' thin layer of radionuclide on the trabecular surfaces. Uniform volume depositions of 45C and 32P were produced by neutron activation and the 14C volume distribution was successfully achieved by rehydrating the bone matrix with 14C-labelled urea solution. Surface depositions of isotope were attempted by introducing a radionuclide into solution. Surface depositions of isotope were attempted by introducing a radionuclide into solution which was in equilibrium with trabecular bone and depositions approximating to an 'infinitely' thin layer of isotope were achieved for 32P and 14C using adenosine triphosphate-labelled molecules. Ionic calcium penetrates the bone matrix very rapidly and the best 45Ca surface deposition that could be produced penetrated the trabecular surface to an average depth of about 10 micrometer. The uniformity of deposition was analysed by autoradiography and the thickness of the 'surface' layer of radionuclide was estimated using microdensitometer scans across selected strands of the autoradiograph and comparing the film density profiles thus produced with calculations.


Activation-induced deaminase-coupled DNA demethylation is not crucial for the generation of induced pluripotent stem cells.
DNA methylation constitutes a major obstacle in the reprogramming of cells to pluripotency. Although little is known regarding the molecular mechanisms of DNA demethylation, activation-induced deaminase (AID), which is known to function in antibody diversification, has been implicated in DNA demethylation through a base excision repair (BER)-mediated pathway. Here we comprehensively examine the plausibility of coupled AID-BER demethylation in the generation of induced pluripotent stem cells (iPSCs) and show that AID is dispensable for reprogramming cells into iPSCs. Additionally, the overexpression of AID and other factors involved in AID-coupled DNA demethylation does not increase the efficiency of reprogramming. Moreover, BER is not likely to play a role in this process. Our results indicate that the reactivation of key genes governing the pluripotency circuitry occurs through a mechanism that is independent of deamination-coupled demethylation.


The COVID-19 pandemic first wave in Sweden: A national registry study of the effects on pediatric anesthesia and surgery.
The COVID-19 pandemic is pushing healthcare systems to their limits. Dramatic reductions in the adult elective surgery are ubiquitous, but corresponding changes in pediatric services are not well described. The Swedish Perioperative Registry contains data on all anesthetic procedures in Sweden, and therefore, provides a unique opportunity to analyze the effect of the pandemic on the pediatric anesthesia capacity on a national level. We hypothesized that there would be a significant reduction in pediatric elective procedures. The aim was to determine the effects on pediatric surgical and anesthetic services during the first wave of the COVID-19 pandemic in Sweden. For this retrospective registry cohort study, we extracted all procedures performed on patients <18 years of age in 2020 and 2019. Weeks 12 to 26 of 2020 were defined as the first wave, and data were analyzed according to level of care, type of surgery, procedure code, and emergency or elective surgery. We found 7015 fewer procedures during the first wave epoch. Elective cases were reduced by 53.7% while emergency surgery was not significantly affected. During the peak of the first wave in April, there was a 72.8% reduction in elective cases; ENT/maxillofacial surgery showed the greatest reduction (86.7%). The surgical and anesthesia capacity recovered to near-normal levels by the end of June 2020. We conclude that the impact of COVID-19 on pediatric surgical procedures in Sweden during the first wave of the pandemic was dramatic, but elective services were restored a few months after the peak.


Determinants of Lifestyle Choices Among South Korean College Students: An Experimental Analysis.
Based on theoretical predictions from human capital theory, the aim of this exploratory study is to analyze the relationship between experimentally elicited, incentivized economic preference parameters, Big Five and Grit personality traits, cognitive ability, and the Alameda Seven lifestyles: smoking, drinking excessively, being overweight or obese, experiencing stress, following a healthy diet, exercising, and sleeping enough. In addition, self-reported general physical and mental health are analyzed. Data collection took place in an incentivized paper-and-pencil experiment. Undergraduate students were recruited as subjects, with the requirement that they be freshmen, but no restrictions based on gender, major, or age. A pre-test and three experimental sessions were conducted in March and in April 2018, with a total of 178 subjects participating. After deleting observations with missing values, a total of n = 138 subjects remained. Regression analysis (multivariate probit models, resulting marginal effects and changes in predicted probabilities of adopting a lifestyle for "ideal types") was used to analyze and compare the determinants of the Alameda Seven health behaviors. Findings suggest that preference parameters, a measure of cognitive ability, and Big Five personality traits are statistically significantly estimated in regressions for the determinants of adopting Alameda Seven health behaviors, but the Grit score is only statistically significantly estimated in a regression for the determinants of reported better general mental health. The findings regarding preference parameters partly lend support to predictions from human capital theory and partly confirm previous findings in health psychology. Those results might be explained by the relatively small sample size and the inclusion of a measure of cognitive ability in addition to personality traits and preferences.


[Effects of different grasses cultivation on apple orchard soil microbial community].
Taking naturally growing grass (NG) as comparison, high-throughput sequencing technique was adopted to examine the effects of Lolium perenne (LP), Trifolium repens (TR), Vulpia myuros (VM), and Vicia villosa (VV) cultivation on soil microbial community in apple orchard of Yanggu County, Shandong Province, and the correlations of soil microbial community with soil nutrient and physicochemical property. The results showed that LP increased the relative abundance of Alternaria and Aspergillus in soil, TR increased that of Gibberella, VM increased Aureobasidium and Chaetomium, and VV increased probiotics such as Nitrososophaera and Metarhizium. However, the quantity of pathogenic bacteria Ralstonia was improved in NG treatment. Results from Spearman correlation analysis showed that the abundance of Bacillus was significantly positively correlated with the concentration of soil organic matter. The abundance of Aspergillus was significantly negatively correlated with available phosphorus, available potassium, CEC, and organic matter. The abundance of Metarhizium was significantly positively correlated with soil pH and CEC. Planting VV between rows of apple orchards, and enhancing the soil organic matter and nutrients content in a certain extent, helped to improve soil micro-ecological environment.


Multimodal spatiotemporal monitoring of basal stem cell-derived organoids reveals progression of olfactory dysfunction in Alzheimer's disease.
Olfactory dysfunction (OD) is a highly prevalent symptom and an early sign of neurodegenerative diseases in humans. However, the roles of peripheral olfactory system in disease progression and the mechanisms behind neurodegeneration remain to be studied. Olfactory epithelium (OE) organoid is an ideal model to study pathophysiology in vitro, yet the reliance on 3D culture condition limits continual in situ monitoring of organoid development. Here, we combined impedance biosensors and live imaging for real-time spatiotemporal analysis of OE organoids morphological and physiological features during Alzheimer's disease (AD) progression. The impedance measurements showed that organoids generated from basal stem cells of APP/PS1 transgenic mice had lower proliferation rate than that from wild-type mice. In concert with the biosensor measurements, live imaging enabled to visualize the spatial and temporal dynamics of organoid morphology. Abnormal protein aggregation and accumulation, including amyloid plaques and neurofibrillary tangles, was found in AD organoids and increased as disease progressed. This multimodal in situ bioelectrical measurement and imaging provide a new platform for investigating onset mechanisms of OD, which would shed new light on early diagnosis and treatment of neurodegenerative disease.


Drug nanocarriers and functional nanoparticles: applications in cancer therapy.
Extending on recent progress in control of matter at the nanoscale, functional nanoparticles and drug nanocarriers are being actively developed with the promise of improved delivery of therapeutic agents. Nanotechnology-enabled drug delivery approaches usually aim at reducing the systemic distribution and associated side-effects typically observed with conventional chemotherapeutic molecules as well as increasing the therapeutic index of the active agents. The enhanced permeation and retention effect has been exploited with notable success to passively deliver nanostructured drug carriers and therapeutic nanoparticles in vivo. Strategies aimed at further enhancing the nanocarriers accumulation within tumor tissues are under intensive investigation and could enable, in a not too distant future, the realization of the long sought after goal of chemotherapy without - or with reduced - side-effects. In particular, immunotargeted approaches are widely predicted to offer much greater specificity towards the target cells and tissues. Advancement in molecular biology has indeed made a wealth of information available in biological processes and application of this new knowledge to nanostructured agents, has also generated unprecedented hope of novel molecular diagnostic and therapeutic strategies. Focusing on oncological applications, this review aims to discuss the recent developments in immunotargeting of functional nanoparticles and drug nanocarriers and the integration of these systems into clinically relevant therapeutic applications.


Development and current status of unconventional platinum anticancer complexes.
Cisplatin is routinely employed for the treatment of testicular, ovarian cancer and head/neck tumors. Typical doses administrated to patients are 100 mg/day for up to five days. It is believed that the mechanism of action appears to be the binding of cis-Pt(NH3)(2) unit to DNA at two neighboring guanine bases. In the years following the introduction of cisplatin, the design of new platinum anticancer drugs concentrated mainly on direct cisplatin analogies, which stuck to the set of structure-activity relationships summarized by Clear and Hoeschele in 1973. Lately, some pioneering strategies towards the synthesis of novel platinum anticancer drugs based on the improved understanding of the mechanism of platinum resistance have emerged. Those are based on either changing the coordinated nitrogen ligand or altering the leaving groups. Other strategies have been shifted to discover "non classical" drugs that can act in a way different from cisplatin. Abnormal structures that violate the empirical structure-activity relationships of platinum compounds and multinuclear complexes are examples of these compounds. Several review articles appeared during recent years dealing with the synthesis, preclinical screening, and mechanism of action of platinum-based anticancer drugs. In this review, the progress in the field of anticancer chemistry based on unconventional platinum antitumor agents during the last 10 years will be highlighted. Most of the complexes that illustrate the recent and the previous prominent strategies will be presented.


High hydrostatic pressure as a method to preserve fresh-cut Hachiya persimmons: A structural approach.
The "Hachiya" persimmon is the most common astringent cultivar grown in California and it is rich in tannins and carotenoids. Changes in the microstructure and some physicochemical properties during high hydrostatic pressure processing (200-400 MPa, 3 min, 25 ℃) and subsequent refrigerated storage were analyzed in this study in order to evaluate the suitability of this non-thermal technology for preservation of fresh-cut Hachiya persimmons. The effects of high-hydrostatic pressure treatment on the integrity and location of carotenoids and tannins during storage were also analyzed. Significant changes, in particular diffusion of soluble compounds which were released as a result of cell wall and membrane damage, were followed using confocal microscopy. The high-hydrostatic pressure process also induced changes in physicochemical properties, e.g. electrolyte leakage, texture, total soluble solids, pH and color, which were a function of the amount of applied hydrostatic pressure and may affect the consumer acceptance of the product. Nevertheless, the results indicate that the application of 200 MPa could be a suitable preservation treatment for Hachiya persimmon. This treatment seems to improve carotenoid extractability and tannin polymerization, which could improve functionality and remove astringency of the fruit, respectively.


[Efferent projections of the caudate nucleus to the cerebral cortex in monkeys].
The efferent direct connections of the head and tail of the caudate nucleus with the cortex of the ipsilateral hemisphere were studied in adult Macaca mulatta. Degeneration of endings in the frontal, precentral and postcentral convolutions was found ipsilaterally in sections of the brain impregnanted after Wiitanen after destruction of the caudate nucleus. The course of solitary fibres to sites of localization of degenerating endings was followed. as compared with the cat in which caudate-cortical projections had been established earlier, cortical zones with degenerated endings were located more discretely in Macaca mulatta.


High frequency ventilation.
What is high frequency ventilation? Is it useful or just another transatlantic fad? Apart from appearing to break all the rules of physiology, this technique has definite advantages over conventional ventilation in certain areas and great potential in others.


Strain-specific susceptibility for pulmonary metastasis of sarcoma 180 cells in inbred mice.
Most cancer deaths are a result of metastasis. To extend our understanding of the factors that influence the process, we aimed to develop a mouse model of pulmonary metastasis that can be assayed in multiple inbred mouse strains for further use in identification of host genetic variants that influence metastasis. We used i.v. injection of Sarcoma 180 (S180) cells, which can be tracked and quantified by bioluminescence imaging. We observed growth of S180 cells solely in the lung and observed a wide range of pulmonary metastasis among inbred mouse strains. Interestingly, we noted that the BTBRT+tf/J strain exhibited complete clearance and provide evidence that the mechanism of resistance may involve immune factors, as strains subjected to whole-body irradiation are significantly more susceptible to tumor growth. One possible mechanism of resistance to pulmonary metastasis in BTBRT+tf/J mice may require T-cell function. Our experiments present a new mouse model for further characterization of the genetics and mechanisms of pulmonary metastasis.


Differences in Cardiovascular Regulation to Head-up Tilt between Healthy and Hypertensive Subjects.
To date there have only been limited studies exploring abnormal hemodynamic responses to head-up tilt tests (HUTs) in elderly, treated patients with hypertension. Cardiovascular regulation in response to HUT as well as upright hemodynamics may be altered when older hypertensive patients with antihypertensive treatments are studied. Hypertensive patients with and without receiving antihypertensive medication and above the age of 45 were recruited in this study. This study compared the cardiovascular responses to HUT and at rest between healthy and hypertensives using non-invasive hemodynamic measurements. Parameters such as systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate (HR), stroke index (SI) and total peripheral resistance index (TPRI) were measured in 40 subjects (20 healthy and 20 hypertensives) for 10-min supine baseline, 10-min HUT at 70◦ and 6-min supine recovery. At rest and during HUT, SBP and TPRI were significantly higher in hypertensives together with a significantly smaller baseline SI. In response to HUT, both groups showed changes in hemodynamic parameters at differing degrees. During recovery, all parameters returned to the baseline range. Our findings indicated that hypertensive patients of older age being treated by antihypertensive drugs may have different cardiovascular changes in response to orthostatic stress.Clinical Relevance- This pilot study describes how cardiovascular regulation in response to postural change may behave differently in hypertensive elder patients taking antihypertensive drugs.


Supervision, scheduling, satisfaction and shared working: how experiences of junior doctors relate to excess mortality within the NHS.
We sought to explore associations between trainee doctor perception and excess patient mortality. Data from two publicly available databases reflecting mortality and components of trainee satisfaction within 81 NHS healthcare institutions between the years 2012 and 2019 were analysed. Pearson's correlation coefficients were calculated. All domains of trainee perception were correlated with excess mortality. Clinical supervision out of hours (R=-0.44; p<0.0001), teamwork (R=-0.36; p<0.0001) and clinical supervision at any time (R=-0.35; p<0.0001) were most strongly correlated. Most associations remained consistent year on year. Trainee doctor perceptions of clinical supervision, rota design and teamwork within the NHS are consistently correlated with excess patient mortality. Further exploration of these associations could identify opportunities for interventions to reduce excess patient mortality. Given the clinical significance of our findings, organisations should consider rapid implementation of evidence-based interventions where they exist.


N-terminal domains of DELLA proteins are intrinsically unstructured in the absence of interaction with GID1/gibberellic acid receptors.
The plant growth-repressing DELLA proteins (DELLAs) are known to represent a convergence point in integration of multiple developmental and environmental signals in planta, one of which is hormone gibberellic acid (GA). Binding of the liganded GA receptor (GID1/GA) to the N-terminal domain of DELLAs is required for GA-induced degradation of DELLAs via the ubiquitin-proteasome pathway, thus derepressing plant growth. However, the conformational changes of DELLAs upon binding to GID1/GA, which are the key to understanding the precise mechanism of GID1/GA-mediated degradation of DELLAs, remain unclear. Using biophysical, biochemical, and bioinformatics approaches, we demonstrated for the first time that the unbound N-terminal domains of DELLAs are intrinsically unstructured proteins under physiological conditions. Within the intrinsically disordered N-terminal domain of DELLAs, we have identified several molecular recognition features, sequences known to undergo disorder-to-order transitions upon binding to interacting proteins in intrinsically unstructured proteins. In accordance with the molecular recognition feature analyses, we have observed the binding-induced folding of N-terminal domains of DELLAs upon interaction with AtGID1/GA. Our results also indicate that DELLA proteins can be divided into two subgroups in terms of their molecular compactness and their interactions with monoclonal antibodies.


Fibromyalgia syndrome. An emerging but controversial condition.
The clinical manifestations, laboratory findings, and treatment results of 118 patients with fibromyalgia followed up by one investigator were compared with those of other recent reports. The history of this syndrome and recent efforts to establish diagnostic criteria and to understand underlying pathophysiologic mechanisms were studied. A practical, noninvasive office-based evaluation and conservative treatment approach were developed, determined by an understanding of the natural history of this common but controversial disorder.


Postoperative 30-day mortality following surgical resection for colorectal cancer in veterans: changes in the right direction.
Temporal changes in short-term mortality following surgery for colorectal cancer (CRC) are unknown. We examined temporal changes in 30-day postoperative mortality, as well as changes in preoperative and postoperative disorders that could contribute to 30-day mortality. Using national Veterans Administration (VA) administrative data, we identified patients with CRC during 1987-2000 who received surgical resection. Cox proportional hazards models were used to evaluate the association between the risk of 30-day mortality and year of surgical resection, while adjusting for several preoperative disorders, disease comorbidity, as well as hospital surgical volume. A total of 32,621 patients were identified. The 30-day postoperative mortality declined from 4.7% during 1987-1988 to 3.9% during 1998-2000. Patients who received surgical resection during 1992-1994, 1995-1997, and 1998-2000 had a 14, 14, and 27% lower adjusted risk of 30-day mortality, respectively, compared with those resected in 1987-1988. Preoperative disorders associated with increased mortality included chronic pulmonary disease, congestive heart failure, diabetes, hemiplegia/paraplegia, moderate/severe liver disease, and renal disease. Significant declines were observed in several postoperative disorders including anesthesia complications and thromboembolism. An improvement in 30-day postoperative mortality following surgical resection for CRC was observed. Declining preoperative and postoperative disorders, as well as improvements in surgical care, could partly explain these findings.


RAADS-14 Screen: validity of a screening tool for autism spectrum disorder in an adult psychiatric population.
Autism spectrum disorder (ASD) can be difficult to distinguish from other psychiatric disorders. The clinical assessment of ASD is lengthy, and has to be performed by a specialized clinician. Therefore, a screening instrument to aid in the identification of patients who may have undiagnosed ASD should be useful. The purpose of this study was to develop such a screening instrument. Based on the 80 item Ritvo Autism and Asperger Diagnostic Scale-Revised (RAADS-R), we developed a 14 item self-evaluation questionnaire, the RAADS-14 Screen. In total, 135 adults with ASD and 508 psychiatric controls completed the abridged version of the RAADS-R. The RAADS-14 Screen score was significantly higher in the ASD group than in the control samples, with a median score of 32 for ASD, 15 for attention deficit hyperactivity disorder, and 11 for other psychiatric disorders (P < 0.001). A cut-off score of 14 or above reached a sensitivity of 97% and a specificity of 46 to 64%. A factor analysis identified three factors consistent with mentalizing deficits, social anxiety, and sensory reactivity relevant for the diagnosis of ASD. The psychometric properties of RAADS-14 Screen were shown to be satisfactory. The results of this study indicate that RAADS-14 Screen is a promising measure in screening for ASD in adult psychiatric outpatients.


[Nutrition and dietary recommendations for the elderly "Public Health" Working Group of the Spanish Nutrition Society].
With the increase of life expectancy, the desire to maintain good health, functionality and maximum quality of life at advanced ages, for which nutrition plays a critical role, is a priority for the elderly. Though genetic factors are a determinant of life expectancy, there are several extrinsic factors which have a great influence on the quality of life of the elderly. Diet and nutritional status have a great influence, especially in the prevention and treatment of several diseases, which affect this heterogeneous and vulnerable age group. The nutritional status and needs of elderly people are associated with age-related biological, psychological and often socio-economic changes. All of these changes can increase the risk of developing a number of age-related diseases. In developed countries the elderly are the most affected by malnutrition, either because of a deficiency (energy and several nutrients) or an excess, leading to obesity and related diseases. This review highlights the most important factors affecting nutritional status in elderly people and focus on the need to maintain adequate physical activity level and an optimal physic, psychic and social functional capacity. It discusses dietary reference intakes and guidelines to improve and/or maintain adequate nutritional status in older people in order to reduce susceptibility to some illness and disease.


Hydrocephalus and hypothalamic involvement in pediatric patients with craniopharyngioma or cysts of Rathke's pouch: impact on long-term prognosis.
Pediatric patients with sellar masses such as craniopharyngioma (CP) or cyst of Rathke's pouch (CRP) frequently suffer disease- and treatment-related sequelae. We analyzed the impact and prognostic relevance of initial hydrocephalus (HY) and hypothalamic involvement (HI) on long-term survival and functional capacity (FC) in children with CP or CRP. Using retrospective analysis of patient records, presence of initial HY or HI was assessed in 177 pediatric patients (163 CP and 14 CRP). Twenty-year overall survival (OS) and progression-free survival (PFS), FC, and BMI were analyzed with regard to initial HY, degree of resection, or HI. Of the 177 patients, 105 patients (103/163 CP and 2/14 CRP) presented with initial HY and 96 presented with HI. HY at diagnosis was associated (P=0.000) with papilledema, neurological deficits, and higher BMI at diagnosis and during follow-up. OS, PFS, and FC were not affected by HY at initial diagnosis. HI at diagnosis (96/177) had major negative impact on long-term prognosis. Sellar masses with HI were associated with lower OS (0.84±0.04; P=0.021), lower FC (P=0.003), and higher BMI at diagnosis and last follow-up (P=0.000) when compared with sellar masses without HI (OS: 0.94±0.05). PFS was not affected by HI or degree of resection. Initial HY has no impact on outcome in patients with sellar masses. OS and FC are impaired in survivors presenting with initial HI. PFS is not affected by HY, HI, or degree of resection. Accordingly, gross-total resection is not recommended in sellar masses with initial HI to prevent further hypothalamic damage.