Nitric oxide: a signal for ADP-ribosylation of proteins.
Nitric oxide (NO), a highly reactive gas, is now established as a major messenger molecule regulating blood vessel dilation, immune functions and serving as a neurotransmitter in brain and peripheral nervous system. NO can also act as a tumoricidal and bactericidal molecule. The effect of NO to dilate blood vessels is largely explained by stimulation of soluble guanylate cyclase (a heme-iron containing protein) leading to formation of cGMP and protein phosphorylation. This is considered to be the main physiological signaling mechanism of NO. NO also binds to non-heme iron-containing proteins and this has been considered as a pathophysiological or cytotoxic action of NO. Furthermore, NO, more correctly nitrosonium (NO+) which can be formed by the removal of one electron, reacts with protein SH-groups to cause the S-nitrosylation of proteins. We have recently established a link between NO and the S-nitrosylation and mono-ADP-ribosylation of the enzyme glyceraldehyde 3-monophosphate dehydrogenase, which adds a further protein modification mechanism for NO action. This links the formation of the second messenger molecule NO to post-translational protein modification and adds a new dimension to NO in the communication of intracellular signals.


A case of "myopathy with tubular aggregates" with increased muscle fibre sensitivity to caffeine.
A 23-year-old man with "myopathy with tubular aggregates" had suffered from exercise-induced muscle cramps for 1 year. His general and neurological findings were normal. Laboratory investigations were within normal limits except for a slightly elevated serum creatine kinase level. Muscle biopsy showed some small angular fibres and scattered type 2B fibres with prominent tubular aggregates originating from the sarcoplasmic reticulum. Since the muscle fibres contracted at a lower concentration of caffeine, increased muscle fibre sensitivity to caffeine is probably related to muscle cramps in this disorder. Tubular aggregates are then secondarily formed in the muscle fibres.


Modulation of NAC transcription factor NST1 activity by XYLEM NAC DOMAIN1 regulates secondary cell wall formation in Arabidopsis.
In Arabidopsis, secondary cell walls (SCW) are formed in fiber cells and vessel cells in vascular tissue for providing plants with mechanical strength and channels for the long distance transportation of water and nutrients. NAC SECONDARY WALL THICKENING PROMOTING FACTOR1 (NST1) acts as a key gene for the initiation of SCW formation through a hierarchical transcription network. In this study, we report that NST activity is modulated by the NAC domain transcription factor XYLEM NAC DOMAIN1 (XND1) during plant growth. Using yeast two-hybrid screening and in vivo protein interaction analysis, XND1 was identified as an NST-interacting protein that modulates NST1 activity. XND1 and NST1 were co-localized in the nucleus and the interaction of XND1 with NST1 resulted in inhibition of NST1 transactivation activity. In the process of inflorescence growth, XND1 was expressed with a similar pattern to NST1. Up-regulation of XND1 in fiber cells repressed SCW formation. The study demonstrates that NST1 activity is modulated by XND1 in the regulation of secondary cell walls formation.


Septal myectomy for obstructive hypertrophic cardiomyopathy in pediatric patients: early and late results.
Symptomatic pediatric patients with obstructive hypertrophic cardiomyopathy (HCM) have a higher death rate (6% annually) compared with adults. Transaortic left ventricular septal myectomy provides excellent outcomes for adults with obstructive HCM. We sought to assess the effect of septal myectomy on late survival and outcome in pediatric patients with obstructive HCM. From 1975 to 2003, 56 pediatric patients underwent septal myectomy for obstructive HCM. Mean age at diagnosis was 6.3 +/- 5.4 years. Ages at operation ranged from 2 months to 20 years (mean 11 +/- 5.6 years). Concomitant procedures included mitral valve repair (n = 7), closure of atrial septal defect (n = 3), and other (n = 5). After myectomy, mean left ventricular outflow tract (LVOT) gradient decreased from 103 +/- 34 to 16 +/- 12 mm Hg and mean degree of mitral regurgitation decreased from 2.0 +/- 1.0 to 1.0 +/- 0.3 (both p < 0.0001). There were no early deaths. Four patients underwent elective cardioverter defibrillator implantation and 1 patient received a permanent pacemaker. Follow-up ranged up to 29 years (mean, 8.6 +/- 6.2). Cardiac reoperations were required in 8 patients: heart transplantation (n = 2), repeat myectomy (n = 2), mitral valve repair-replacement (n = 2), Konno-Rastan procedure (n = 1), and aortic valve replacement (n = 1). Age 14 years or less at operation was the only predictor of late reoperation (p = 0.017). Two patients died late; one suddenly without residual LVOT obstruction and one from chronic rejection after heart transplantation. Ninety-six percent of surviving patients were in New York Heart Association functional class I or II. Survival estimates at 5 and 10 years were 97% and 93%, respectively. Septal myectomy is safe and effective in symptomatic pediatric patients with obstructive HCM. Late survivorship compares very favorably with the natural history of this disease.


Fighting for Fat: Gluttonous Tumors and Starved T Cells.
In this issue of Cell, Ringel et al. reveal a link between lipid utilization in the tumor microenvironment and anti-tumor immunity in obese mice. These findings provide one explanation for how obesity worsens cancer outcomes and may point to a new metabolic approach to treating some cancers.


Radical vulvectomy and "sneak" superficial inguinal lymphadenectomy with a single elliptic incision.
With a single elliptic radical vulvectomy incision, resection of the primary tumor and bilateral superficial inguinal lymphadenectomies were performed in 18 patients with invasive squamous cell carcinoma of the vulva. Twelve patients (67%) had no morbidity from the procedure, while five patients (28%) developed significant wound breakdown and one patient (5%) developed a groin hematoma. For select patients with invasive vulvar cancer, this modification of the standard "butterfly" incision offers an acceptable surgical approach to this disease, allowing removal of the primary lesion and adequate sampling of the groin lymphatics. Results of pathologic examinations allow selection of those patients requiring further therapy.


Identification of nontuberculous mycobacterial infection by IS6110 and hsp65 gene analysis on lung tissues.
The clinical, histologic, and radiographic presentations of nontuberculous mycobacterial (NTM) lung disease are usually indistinguishable from those of reactivated pulmonary tuberculosis (TB), so it remains a great challenge for the clinician to make treatment decisions for patients with old TB and a positive culture result for NTM. This study investigated whether the mycobacterial specific heat shock protein 65 (hsp65) and Mycobacterium tuberculosis (MTB)-specific IS6110 gene would present in pulmonary lesions of patients with NTM pulmonary infection. Formalin-fixed and paraffin-embedded (FFPE) tissue blocks of 24 patients with NTM infections treated at the hospital from 1998 to 2008 were included. Mycobacterial hsp65 gene was amplified in 20 of the 24 patients, and the species identified by sequencing was consistent with corresponding culture results in 12 of these patients. MTB-specific IS6110 gene was detected in 3 of the 7 patients who had old TB and a subsequent diagnosis of fibrocavitary NTM lung disease. Polymerase chain reaction (PCR) analysis of hsp65 gene also confirmed the presence of MTB genes in 2 of these 3 patients. Our results indicate that PCR amplification and sequencing of the mycobacterial hsp65 gene is a sensitive assay for identification of NTM species in FFPE materials. However, consistent results of PCR analysis, microbiology study, histologic manifestations, radiology, and clinical presentation are important for correct diagnosis of NTM pulmonary infection. The presence of MTB gene in patients with fibrocavitary NTM lung lesions poses a clinical dilemma for deciding concurrent treatment TB and NTM infection.


Comparison of Percutaneous Renal Mass Biopsy and R.E.N.A.L. Nephrometry Score Nomograms for Determining Benign Vs Malignant Disease and Low-risk Vs High-risk Renal Tumors.
To compare the accuracies of renal mass biopsy (RMB) and R.E.N.A.L. nephrometry score (RNS) nomograms for predicting benign vs malignant disease, and low- vs high-risk renal tumors. We included 281 renal masses in 277 patients who had complete RNS, preoperative RMB, and final pathology from renal surgery for clinically localized renal tumors. RMB and final pathology were determined to be benign or malignant, and malignancies were classified as low-risk (Fuhrman grade I/II) or high-risk (Fuhrman grade III/IV) (benign included in low-risk group). Previously published RNS nomograms were used to determine probabilities of any cancer and high-risk cancer. The gamma statistic was used to assess strength of association between RMB or RNS with final pathology. Of the 281 masses, 13 (5%) and 268 (95%) were confirmed benign and malignant, respectively, and 155 (55%) and 126 (45%) were confirmed low-risk and high-risk, respectively, on final pathology. The areas under the curve of the RNS nomograms for benign vs malignant disease and for low-risk vs high-risk renal tumors were 0.56 and 0.64, respectively. Concordances for predicting benign vs malignant disease were 99% for RMB (P < .01, gamma 0.99) and 29% for RNS nomogram (P = .16, gamma 0.38). Concordances for predicting low-risk vs high-risk renal tumors were 67% for RMB (P < .01, gamma 0.97) and 61% for RNS nomogram (P < .01, gamma 0.47), respectively. Although RNS nomograms are useful for discriminating between benign vs malignant renal masses, and low-risk vs high-risk renal tumors, they are outperformed by RMB.


MPP+-induces PUMA- and p53-dependent, but ATF3-independent cell death.
Parkinson's disease (PD) is characterized by the progressive loss of dopaminergic neurons in the substantia nigra (SN) and depletion of striatal dopamine (DA), leading to a range of motor symptoms, including resting tremor, rigidity, bradykinesia and postural abnormalities. The neurotoxin (MPTP) and its active metabolite, 1-methyl-4-phenylpyridinium (MPP(+)), cause dopaminergic cell loss in a variety of animal species and produce symptoms similar to those seen in PD. Our lab has shown that MPP(+) activates cell stress pathways, including the unfolded protein response (UPR) in mouse primary mesencephalic cultures. The BH3-only protein, PUMA (p53 upregulated mediator of apoptosis), has been shown to be activated in response to many cellular stresses, including endoplasmic reticulum (ER) stress and UPR, and to induce cell death. Therefore, we hypothesized that PUMA may mediate MPP(+) toxicity. To test this hypothesis, we compared the response of primary mesencephalic cultures from wild-type and PUMA deficient (-/-) mice to MPP(+). We also utilized cultures from p53 -/- and activating transcription factor 3 (ATF3) -/- mice to further elucidate the pathways involved. These studies revealed that PUMA and p53, but not ATF3, are required for MPP(+)-induced cell death, suggesting that UPR activation is parallel to the induction of MPP(+)-induced cell death.


9-(5-Carboxypentylamino)-acridine. An affinity adsorbent ligand for Electrophorus acetylcholinesterase.
We have developed a simple method for the synthesis of the ligand 9-(5-carboxypentylamino)-acridine and the resulting affinity adsorbent using Sepharose CL-4B. This affinity adsorbent is efficient and specific for purification acetylcholinesterase (acetylcholine acetylhydrolase, EC 188.8.131.52) from Electrophorus electricus electric organ.


Lost after translation: post-translational modifications by bacterial type III effectors.
Many Gram-negative bacterial pathogens use the type III secretion system to deliver effector proteins into host cells. These effectors use various mechanisms to exploit host processes to the advantage of the pathogen. A large group of effectors use post-translational modifications, either reversible or irreversible, to manipulate host proteins, and while most of these mechanisms mimic eukaryotic activities, others appear to be unique biochemical functions. Deciphering such mechanisms and identifying the host targets of these effectors sheds light on eukaryotic signaling pathways and immune responses.


MicroRNA-27b targets CBFB to inhibit differentiation of human bone marrow mesenchymal stem cells into hypertrophic chondrocytes.
Human bone marrow-derived mesenchymal stem cells (hBMSCs) have chondrocyte differentiation potential and are considered to be a cell source for cell-transplantation-mediated repair of cartilage defects, including those associated with osteoarthritis (OA). However, chondrocyte hypertrophic differentiation is a major obstacle for the application of hBMSCs in articular cartilage defect treatment. We have previously shown that microRNA-27b (miR-27b) inhibits hypertrophy of chondrocytes from rat knee cartilage. In this study, we investigated the role of miR-27b in chondrocyte hypertrophic differentiation of hBMSCs. Chondrogenic marker and microRNA expression in hBMSC chondrogenic pellets were evaluated using RT-qPCR and immunohistochemistry. The hBMSCs were transfected with miR-27b before inducing differentiation. Gene and protein expression levels were analyzed using RT-qPCR and western blot. Coimmunoprecipitation was used to confirm interaction between CBFB and RUNX2. Luciferase reporter assays were used to demonstrate that CBFB is a miR-27b target. Chondrogenic differentiation was evaluated in hBMSCs treated with shRNA targeting CBFB. Chondrogenic hBMSC pellets overexpressing miR-27b were implanted into cartilage lesions in model rats; therapeutic effects were assessed based on histology and immunohistochemistry. The hBMSCs showed typical MSC differentiation potentials. During chondrogenic differentiation, collagen 2 and 10 (COL2 and COL10), SOX9, and RUNX2 expression was upregulated. Expression of miR-140, miR-143, and miR-181a increased over time, whereas miR-27b and miR-221 were downregulated. Cartilage derived from hBMSC and overexpressing miR-27b exhibited higher expression of COL2 and SOX9, but lower expression of COL10, RUNX2, and CBFB than did the control cartilage. CBFB and RUNX2 formed a complex, and CBFB was identified as a novel miR-27b target. CBFB knockdown by shRNA during hBMSC chondrogenic differentiation led to significantly increased COL2 and SOX9 expression and decreased COL10 expression. Finally, miR-27b-overexpressing hBMSC chondrogenic pellets had better hyaline cartilage morphology and reduced expression of hypertrophic markers and tend to increase repair efficacy in vivo. MiR-27b plays an important role in preventing hypertrophic chondrogenesis of hBMSCs by targeting CBFB and is essential for maintaining a hyaline cartilage state. This study provides new insights into the mechanism of hBMSC chondrocyte differentiation and will aid in the development of strategies for treating cartilage injury based on hBMSC transplantation.


Constitutive Activating Eel Luteinizing Hormone Receptors Induce Constitutively Signal Transduction and Inactivating Mutants Impair Biological Activity.
In contrast to the human lutropin receptor (hLHR) and rat LHR (rLHR), very few naturally occurring mutants in other mammalian species have been identified. The present study aimed to delineate the mechanism of signal transduction by three constitutively activating mutants (designated M410T, L469R, and D590Y) and two inactivating mutants (D383N and Y546F) of the eel LHR, known to be naturally occurring in human LHR transmembrane domains. The mutants were constructed and measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary (CHO)-K1 cells. The activating mutant cells expressing eel LHR-M410T, L469R, and D590Y exhibited a 4.0-, 19.1-, and 7.8-fold increase in basal cAMP response without agonist treatment, respectively. However, inactivating mutant cells expressing D417N and Y558F did not completely impaired signal transduction. Specifically, signal transduction in the cells expressing activating mutant L469R was not occurred with a further ligand stimulation, showing that the maximal response exhibited approximately 53% of those of wild type receptor. Our results suggested that the constitutively activating mutants of the eel LHR consistently occurred without agonist treatment. These results provide important information of LHR function in fish and regulation with regard to mutations of highly conserved amino acids in glycoprotein hormone receptors.


MRI characteristics of nodular fasciitis of the musculoskeletal system.
To evaluate MRI imaging appearances of nodular fasciitis in a pathologic-proven series of 29 patients. Review of the orthopedic oncology and pathology databases yielded 51 cases of histologically proven nodular fasciitis. MR imaging was available in 29 patients. Three musculoskeletal radiologists retrospectively reviewed all cases in consensus. Imaging features evaluated included location in the body, size, compartmental localization, relationship to fascia, signal characteristics, enhancement pattern, transcompartmental extension, and osseous and intra-articular involvement. There were 15 male and 14 female patients. Mean age was 33 years (range, 16-59 years). Lesions ranged in size from 1.6 to 9 cm with 84 % of lesions measuring less than 4 cm. Twenty-three lesions were located in the upper arm or shoulder girdle. Nine lesions were subcutaneous in location, nine were intra-muscular, and 11 were inter-muscular. Lesions were consistently ovoid in shape with broad fascial contact. They exhibited internal homogenous low T1 and heterogeneous intermediate T2 signal with surrounding edema and slightly inhomogeneous enhancement. Twelve lesions exhibited central non-enhancing areas. Trans-compartmental spread was demonstrated in nine lesions. Osseous changes were seen in five cases and included extrinsic cortical saucerization, medullary edema, and transcortical osseous invasion. Two lesions demonstrated intra-articular extension. MR imaging features of nodular fasciitis are generally non-specific and can be mistaken for a soft tissue sarcoma. This series, the largest MRI series of musculoskeletal cases in the literature, confirms the predilection of nodular fasciitis for the upper extremity in young adults but also demonstrates that aggressive imaging features such as transcompartmental spread, and osseous and intra-articular involvement may be seen in association with this benign soft tissue lesion.


Postnatal development of the actual and total activity of the branched-chain 2-oxo acid dehydrogenase complex in rat tissues.
Actual and total activities of the branched-chain 2-oxo acid dehydrogenase complex were determined in homogenates of quadriceps muscle, heart, liver, kidney and brain from rats of 0-70 days age. All rat tissues except quadriceps muscle showed a marked increase of total activity between 0 and 21 days, heart and kidney also after weaning. The actual activity rose after birth in liver, kidney and brain and after weaning in liver, kidney and heart. The activity state was always about 100% in liver and varied between 40-60% in kidney and brain, 10-23% in heart and 6-12% in quadriceps muscle. The actual activities measured indicate, that the degradation of branched-chain 2-oxo acids mainly takes place in the liver of the newborn, suckling and young-adult rat.


Response of brain oxygen to therapy correlates with long-term outcome after subarachnoid hemorrhage.
Brain oxygen (PbtO2) monitoring can help guide care of poor-grade aneurysmal subarachnoid hemorrhage (aSAH) patients. The relationship between PbtO2-directed therapy and long-term outcome is unclear. We hypothesized that responsiveness to PbtO2-directed interventions is associated with outcome. Seventy-six aSAH patients who underwent PbtO2 monitoring were included. Long-term outcome [Glasgow Outcome Score-Extended (GOS-E) and modified Rankin Scale (mRS)] was ascertained using the social security death database and structured telephone interviews. Univariate and multivariate regression were used to identify variables that correlated with outcome. Data from 64 patients were analyzed (12 were lost to follow-up). There were 530 episodes of compromised PbtO2 (<20 mmHg) during a total of 7,174 h of monitor time treated with 1,052 interventions. Forty-two patients (66 %) survived to discharge. Median follow-up was 8.5 months (range 0.1-87). At most recent follow-up 35 (55 %) patients were alive, and 28 (44 %) had a favorable outcome (mRS ≤3). In multivariate ordinal regression analysis, only age and response to PbtO2-directed intervention correlated significantly with outcome. Increased age was associated with worse outcome (coeff. 0.8, 95 % CI 0.3-1.3, p = 0.003), and response to PbtO2-directed intervention was associated with improved outcome (coeff. -2.12, 95 % CI -4.0 to -0.26, p = 0.03). Patients with favorable outcomes had a 70 % mean rate of response to PbtO2-directed interventions whereas patients with poor outcomes had a 45 % response rate (p = 0.005). Response to PbtO2-directed intervention is associated with improved long-term functional outcome in aSAH patients.


Otolaryngology Utilization in Patients With Achondroplasia: Results From the CLARITY Study.
To quantify otolaryngologic surgery utilization in patients with achondroplasia, and to identify any changes in utilization over the past four decades. Retrospective cohort study. A retrospective cohort study of 1,374 patients with achondroplasia enrolled in the CLARITY retrospective cohort study at four centers of multi-specialty care for patients with achondroplasia. Otolaryngologic surgeries are presented by birth cohort decade. The main outcomes were number of primary and additional otolaryngologic procedures; age at surgery; likelihood of repeated surgery; temporal trends in surgical utilization. In this cohort of 1,374 patients with achondroplasia, 620 (45.1%) had pharyngeal surgery at least once, 150 (10.9%) had pharyngeal surgery on more than one occasion, and patients who had adenoidectomy first were 2.68 times more likely to require a second pharyngeal surgery than those who had adenotonsillectomy. Seven hundred and seventy-nine (56.7%) had tympanostomy tubes placed at least once, and 447 (32.5%) had tympanostomy tubes placed more than one time. Age at first pharyngeal surgery decreased by 1.2 years per birth cohort decade, and age at tympanostomy tube placement decreased by 1.1 years per decade. Patients with achondroplasia often require otolaryngologic surgery, particularly adenoidectomy and/or tonsillectomy as well as tympanostomy tube placement. Such surgery is performed now more frequently and at younger ages than in earlier decades. While otolaryngologic disease associated with achondroplasia is now recognized earlier and treated more frequently, long-term outcome studies are needed. 3 Laryngoscope, 132:1548-1554, 2022.


Affinity of anti-spike antibodies in SARS-CoV-2 patient plasma and its effect on COVID-19 antibody assays.
Measuring anti-spike protein antibodies in human plasma or serum is commonly used to determine prior exposure to SARS-CoV-2 infection and to assess the anti-viral protection capacity. According to the mass-action law, a lesser concentration of tightly binding antibody can produce the same quantity of antibody-antigen complexes as higher concentrations of lower affinity antibody. Thus, measurements of antibody levels reflect both affinity and concentration. These two fundamental parameters cannot be disentangled in clinical immunoassays, and so produce a bias which depends on the assay format. To determine the apparent affinity of anti-spike protein antibodies, a small number of antigen-coated magnetic microparticles were imaged by fluorescence microscopy after probing antigen-antibody equilibria directly in patient plasma. Direct and indirect anti-SARS-CoV-2 immunoassays were used to measure antibody levels in the blood of infected and immunised individuals. We observed affinity maturation of antibodies in convalescent and vaccinated individuals, showing that higher affinities are achieved much faster by vaccination. We demonstrate that direct and indirect immunoassays for measuring anti-spike protein antibodies depend differently on antibody affinity which, in turn, affects accurate interpretation of the results. Direct immunoassays show substantial antibody affinity dependence. This makes them useful for identifying past SARS-CoV-2 exposure. Indirect immunoassays provide more accurate quantifications of anti-viral antibody levels. The authors are all full-time employees of Abbott Laboratories. Abbott Laboratories provided all operating funds. No external funding sources were used in this study.


Early effect of tacrolimus in improving excitation-contraction coupling in myasthenia gravis.
Tacrolimus (FK506) is a macrolide T-cell immunomodulator used to treat myasthenia gravis (MG). Besides immunosuppression, tacrolimus has been reported to have the potential to increase muscle strength by enhancing ryanodine receptor (RyR) function. However, few attempts have been made to demonstrate the early effect of tacrolimus as an RyR enhancer in clinical investigation. In 20 MG patients, masseteric compound muscle action potential (CMAP) and mandibular movement-related potentials (MRPs) were recorded simultaneously after stimulating the trigeminal motor nerve with a needle electrode. The excitation-contraction (E-C) coupling time (ECCT) was calculated by the latency difference between CMAP and MRP. Bite force was measured using a pressure-sensitive sheet. Serial assessments of % decrement in masseteric repetitive nerve stimulation (RNS), ECCT and bite force were performed before and within 4 weeks of tacrolimus (3 mg day(-1)) treatment. The median (mean, range) interval of assessment was 2 (2.4, 1-4) weeks. We also measured serum antibodies against RyR, acetylcholine receptor and muscle-specific receptor tyrosine kinase. Bite force increased after tacrolimus treatment accompanying clinical improvement assessed by Myasthenia Gravis Foundation of America classification, but the bite force difference did not reach statistical significance. Wilcoxon matched-pairs signed-ranks test detected a significant ECCT shortening in 12 patients assessed after 1-2 weeks of tacrolimus treatment as well as in eight patients assessed after 3-4 weeks. In contrast, masseteric CMAP and % decrement showed no significant changes after short-term tacrolimus treatment. Tacrolimus induces ECCT shortening accompanying clinical improvement despite no improvement in % decrement within 2 weeks. This early effect of tacrolimus may imply a pharmacological enhancement of RyR function to improve E-C coupling in MG.


The effects of high-monosaccharide diets on development and biochemical composition of white-eyed mutant strain of house cricket (Acheta domesticus).
Tryptophan (TRP) is one of the essential amino acids in the animal body. Its exogenicity and low concentrations mean that it can be regarded as one of the key regulatory molecules at the cellular as well as physiological level. It has been shown to have a number of essential functions, such as in the production of other biologically active molecules. The main objective of this project was to investigate the effects of a high monosaccharide diet (HMD) on a hemimetabolic insect-house cricket (Acheta domesticus) and a mutant strain with impaired visual pigment synthesis (closely related to the tryptophan and kynurenine (KYN) metabolic pathway)-white eye. This study was aimed at determining the effects of glucose and fructose on cricket development and biochemical composition. A parallel goal was to compare the response of both cricket strains to HMD. ELISA assays indicated dysfunction of the TRP-KYN pathway in white strain insects and an elevated KYN/TRP ratio. Biochemical analyses demonstrated the effects of HMD mainly on fat and glycogen content. A decrease in food intake was also observed in the groups on HMD. However, no changes in imago body weight and water content were observed. The results of the study indicate a stronger response of the white strain to HMD compared to the wild-type strain. At the same time, a stronger detrimental effect of fructose than of glucose was apparent. Sex was found to be a modulating factor in the response to HMD.


Determination of optimum operating conditions of carmine decoloration by UV/H2O2 using response surface methodology.
In this study, the photolytic decoloration of carmine (C.I. Natural Red 4) via UV radiation in the presence of H2O2 was optimized using response surface methodology (RSM). According to analysis of variance (ANOVA) results, the proposed model can be used to navigate the design space. It was found that the response of carmine degradation is very sensitive to the independent factors of carmine concentration, H2O2 concentration, pH and reaction time. The proposed model for D-optimal design fitted very well with the experimental data with R2 and R(adj)2 correlation coefficients of 0.998 and 0.997, respectively.


Controlled Self-Assembly of λ-DNA Networks with the Synergistic Effect of a DC Electric Field.
Large-scale and morphologically controlled self-assembled λ-DNA networks were successfully constructed by the synergistic effect of a DC electric field. The effect of DNA concentration, direction, and intensity of the electric field, even the modification of the mica surface using Mg2+ on the characteristics of the as-prepared DNA networks, were investigated in detail by atomic force microscopy (AFM). It was found that the horizontal electric field was more advantageous to the formation of DNA networks with more regular structures. At the same concentration, the height of DNA network was not affected significantly by the intensity change of the horizontal electric field. The modification of Mg2+ on mica surface increased the aggregation of DNA molecules, which contributed to the morphological change of the DNA networks. Furthermore, DNA molecules were obviously stretched in both horizontal and vertical electric fields at low DNA concentrations.


Phase I and scintigraphy studies to evaluate safety, tolerability, pharmacokinetics, and lung deposition of inhaled GDC-0214 in healthy volunteers.
Several inflammatory cytokines that promote inflammation and pathogenesis in asthma signal through the Janus kinase 1 (JAK1) pathway. This phase I, randomized, placebo-controlled trial assessed the pharmacokinetics and safety of single and multiple ascending doses up to 15 mg twice daily for 14 days of a JAK1 inhibitor, GDC-0214, in healthy volunteers (HVs; n = 66). Doses were administered with a dry powder, capsule-based inhaler. An accompanying open-label gamma scintigraphy study in HVs examined the lung deposition of a single dose of inhaled Technetium-99m (99m Tc)-radiolabeled GDC-0214. GDC-0214 plasma concentrations were linear and approximately dose-proportional after both single and multiple doses. Peak plasma concentrations occurred at 15-30 min after dosing. The mean apparent elimination half-life ranged from 32 to 56 h across all single and multiple dose cohorts. After single and multiple doses, all adverse events were mild or moderate, and none led to treatment withdrawal. There was no clear evidence of systemic toxicity due to JAK1 inhibition, and systemic exposure was low, with plasma concentrations at least 15-fold less than the plasma protein binding-corrected IC50 of JAK1 at the highest dose. Scintigraphy showed that approximately 50% of the emitted dose of radiolabeled GDC-0214 was deposited in the lungs and was distributed well to the peripheral airways. 99m Tc-radiolabeled GDC-0214 (1 mg) exhibited a mean plasma Cmax similar to that observed in phase I at the same dose level. Overall, inhaled GDC-0214 exhibited pharmacokinetic properties favorable for inhaled administration.


Esophagogastroduodenoscopy Screening Intentions During the COVID-19 Pandemic in Japan: Web-Based Survey.
The number of people undergoing cancer screening decreased during the COVID-19 pandemic. The pandemic may have affected the willingness and motivation of undergoing cancer screening by those eligible for it. This study aims to clarify the effect of the COVID-19 pandemic on the intention to undergo cancer and esophagogastroduodenoscopy (EGD) screening. We performed a web-based survey on the intention to undergo screening among 1236 men and women aged 20-79 years. The numbers of participants by sex and 10-year age groups were equal. The survey was conducted in January 2021, during which the government declared a state of emergency because of the third wave of the COVID-19 pandemic in Japan. Emergency declarations were issued in 11 prefectures among all the 47 prefectures in Japan. In total, 66.1% (817/1236) of the participants felt anxious about undergoing screening due to COVID-19. More women than men were anxious about undergoing screening. By modality, EGD had the highest percentage of participants with anxiety due to COVID-19. Regarding the intention to change the participants' appointment for screening, the most common strategies were to book an appointment for a time during nonpeak hours, postpone the appointment to a later date, and change the mode of transportation. In addition, 35.8% (442/1236) of the participants were willing to cancel this year's screening appointment. Among the 1236 participants, 757 (61.2%) were scheduled for screening in 2020. Of the 757 participants in this subgroup, 68% (n=515) did not change the schedule, 6.1% (n=46) cancelled, and 26% (n=197) made some changes, including changing the appointment date, hospital, or mode of transportation. Among the 296 participants scheduled for EGD screening, 18.9% (n=56) made some changes, 5.7% (n=17) cancelled on their own, and 2.7% (n=8) cancelled on the hospital's order. Based on the previous screening results, the percentage of participants who felt anxious about EGD due to the COVID-19 pandemic was higher in the order of those who had not undergone screening and those who were judged to be in need of further examination in screening but did not visit a hospital for it. In the logistic regression analysis, the factors associated with anxiety about EGD screening due to the COVID-19 pandemic were "viral infection prevention measures," "waiting time," "fees (medical expenses)," "mode of transportation," "worry about my social position if I contracted COVID-19," and "perceived the risk of gastric cancer." However, "residence in declared emergency area" was not associated with EGD anxiety due to COVID-19. Excessive anxiety about COVID-19 may lead to serious outcomes, such as a "decreasing intention to undergo EGD screening," and it is necessary to thoroughly implement infection prevention measures and provide correct information to examinees.


Concerted CMP-dependent [3H]inositol labeling of phosphoinositides and agonist activation of phospholipase C in rat brain cortical membranes.
[3H]Inositol ([3H]Ins) labeling of phosphoinositides was studied in rat brain cortical membranes. [3H]Ins was incorporated into a common lipid pool through both CMP-dependent and independent mechanisms. These are as follows: (1) a reverse reaction catalyzed by phosphatidyl-inositol (PtdIns) synthase, and (2) the reaction performed by the PtdIns headgroup exchange enzyme, respectively. Membrane phosphoinositides prelabeled in either CMP-dependent or independent fashions were hydrolyzed by guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S)- and carbachol-stimulated phospholipase C. Unlike CMP-dependent labeling, however, CMP-independent incorporation of [3H]Ins into lipids was inhibited by 1 mM (0.04%) sodium deoxycholate. Thus, when PtdIns labeling and phospholipase C stimulation were studied in a concerted fashion, [3H]Ins was incorporated into lipids primarily through the PtdIns synthase-catalyzed reaction because of the presence of deoxycholate required to observe carbachol-stimulation of phospholipase C. Little direct breakdown of [3H]PtdIns was detected because production of myo-[3H]inositol 1-monophosphate was minimal and myo-[3H]inositol 1,4-bisphosphate was the predominant product. Although PtdIns labeling and 3H-polyphosphoinositide formation were unaffected by GTP gamma S and carbachol and had no or little lag period, GTP gamma S- and carbachol-stimulated appearance of 3H-Ins phosphates exhibited an appreciable lag (10 min). Also, flux of label from [3H]Ins to 3H-Ins phosphates was restricted to a narrow range of free calcium concentrations (10-300 nM). These results show the concerted activities of PtdIns synthase, PtdIns 4-kinase, and phospholipase C, and constitute a simple assay for guanine nucleotide-dependent agonist stimulation of phospholipase C in a brain membrane system using [3H]Ins as labeled precursor.


Radical prostatectomy readmissions: Causes, risk factors, national rates, & costs.
Readmissions have substantial clinical and financial impacts on the healthcare system. Radical prostatectomy (RP) is considered a standard treatment in the management of clinically localized prostate cancer. Yet, there is a paucity of research evaluating readmissions for RP in a national dataset. Patients with histologically confirmed prostate cancer managed with RP were identified within the 2016 to 2018 Nationwide Readmissions Database. Patient factors, facility factors, and surgical characteristics were evaluated for associations with readmission using univariable and multivariable analyses. A total of 133,727 patients receiving RP were identified. Early (30-day) and late (31-90-day) readmission rates were 4.2% and 1.8% respectively. The most common cause of early readmission was postoperative digestive system complication (10%) and the most common cause of late readmission was septicemia (13%). On multivariable logistic regression, factors associated with both early and late readmission include nonroutine discharge at index (early: OR 1.877, 95% CI 1.667-2.113; late: OR 1.801, 95% CI 1.490-2.183), and circulatory system comorbidity (early: OR 1.29, 95% CI 1.082-1.538); late: OR 1.515, 95% CI 1.157-1.984). Our findings regarding factors associated with readmission provide insight for RP counseling and may inform postoperative care pathways. Elucidation of readmission trends may allow the identification and proactive management of patients at higher risk for readmission.


Molecular and biochemical characterization of a recombinant endoglucanase rCKT3eng, from an extreme halophilic Haloarcula sp. strain CKT3.
Halophilic cellulases are indispensable enzymes of heavy industrial processes as resistant biocatalysts due to high level activity at extreme conditions. In this study, crude cellulase from an extreme halophilic Haloarcula sp. CKT3 was characterized. Then, recombinant expression of putative endo-1,4-β-glucanase gene, of CKT3 strain, in E. coli BL21(DE3) was performed with the aim of obtaining highly pure, active and robust industrial enzyme for such industrial aplications. The crude cellulase had optimal activity (16.9 U/mg) at 70 °C, pH 7.0 and 4 M NaCl exhibiting good thermostability, high pH and halotolerance. Indeed, it is very stable in water-insoluble organic solvents with log Po/w ≥ 2.13 and highly resistant to SDS (10%). Recombinant CKT3eng has a molecular weight of 36.9 kDa and 99% aminoacid identity to endo-l,4-β-D-glucanase from Haloarcula argentinensis. Its 3D structure was predicted using Phyre2 and I-TASSER. rCKT3eng enzyme provided 31.6 U/mg activity at optimal 50 °C, pH 7.0 and 3 M NaCl. In addition to its quite similar stability values and resistance to organic solvents and SDS, rCKT3eng has superiority over crude enzyme with 1.87-fold higher specific activity. Therefore, rCKT3eng offers a promising enzyme for industrial use with its valuable activity and stability in extreme conditions.


The anterior cricoid split in puppies.
The acute and long-term effects of the anterior cricoid split on the subglottis of puppies intubated from 7 to 14 days are documented. The anterior cricoid split acutely increased the intralumenal cricoid surface area in puppies with intubation-induced airway injury. An intense inflammatory response with mucosal ulceration and granulation tissue is elicited by 14 days of intubation by using the canine model of induced subglottic stenosis developed by Supance et al. [19]. When animals intubated for 14 days underwent an anterior cricoid split on day 7, the airway appears essentially normal by day 14. The split cricoid cartilage maintained a 'U' configuration following the procedure. The region of deficient cartilage anteriorly is bridged by fibrous tissue with normal epithelium lining the lumenal surface. Splitting the cricoid cartilage anteriorly increased the intralumenal area and no long term complications resulted from the procedure. The mechanisms by which the anterior cricoid split expands the airway intralumenal area while maintaining airway support have been reviewed.


Human tumor-induced and naturally occurring Treg cells differentially affect NK cells activated by either IL-2 or target cells.
NK cells play a crucial role in the eradication of tumor cells. Naturally occurring (n) Treg cells and induced (i) Treg cells are two distinct Treg subsets. While the interaction of nTreg cells with NK cells has been investigated in the past, the role of tumor iTreg cells in the modulation of NK-cell function remains unclear. Tumor iTreg cells were generated from CD4(+) CD25(-) T cells in the presence of autologous immature DCs, head and neck cancer cells and IL-2, IL-10, and IL-15. The effect of iTreg cells and nTreg cells on the expression of NKG2D, NKp44, CD107a, and IFN-γ by NK cells, as well as NK tumor-cytolytic activity, were investigated. iTreg cells - similar to recombinant TGF-β and nTreg cells - inhibited IL-2-induced activation of NK cells in the absence of target cell contact. Surprisingly, and in contrast to nTreg cells, iTreg cells enhanced NK-cell activity elicited by target cell contact. The cytolytic activity of NK cells activated by iTreg cells was mediated via perforin and FasL. We conclude that tumor iTreg cells inhibited IL-2-mediated NK-cell activity in the absence of target cells, whereas the tumoricidal activity of NK cells was enhanced by iTreg cells. Our data suggest a complex, previously not recognized, differential regulation of human NK activity by iTreg cells in the tumor microenvironment.


Mesenchymal stem cell injection induces cardiac nerve sprouting and increased tenascin expression in a Swine model of myocardial infarction.
Stem Cell Induces Cardiac Nerve Sprouting. Mesenchymal stem cell (MSC) transplantation is a promising technique to improve cardiac function. Whether MSC can increase cardiac nerve density and contribute to the improved cardiac function is unclear. Anterior wall myocardial infarction was created in 16 swine. One month later, 6 swine were given MSC and fresh bone marrow (BM) into infarcted myocardium (MSC group). Four swine were given fresh BM only (BM group), and 6 swine were given culture media (MI-only group). The swine were sacrificed 95.8 +/- 3.5 days after MI. Six normal swine were used as control. Immunocytochemical staining was performed using antibodies against growth-associated protein 43 (GAP43), tyrosine hydroxylase (TH), and three subtypes of tenascin (R, C, and X). Five fields per slide were counted for nerve density. The results show the following. (1). There were more GAP43-positive nerves in the MSC group than in the BM, MI-only, or Control group (P < 0.0001). TH staining showed higher nerve densities in the MSC group than in the MI-only (P < 0.01) or Control group (P < 0.0001) in the atria. (2). There were more sympathetic (TH-positive) nerves in myocardium distant from infarct than in the peri-infarct area (P < 0.05). (3). Optical intensity and color analyses showed significantly higher tenascin R and tenascin C expression in the MSC and BM groups than in the MI-only or Control group (P < 0.01). MSC injected with BM into swine infarct results in overexpression of cardiac tenascin, increased the magnitude of cardiac nerve sprouting in both atria and ventricles, and increased the magnitude of atrial sympathetic hyperinnervation 2 months after injection.


Pathways to depression care: help-seeking experiences of low-income Latinos with diabetes and depression.
This qualitative study examines help-seeking pathways to depression care of low-income Latinos with diabetes and major depression. A purposive sample (N = 19) of Spanish-speaking, immigrant, low-income Latinos was selected from a randomized clinical trial targeting Latinos with diabetes and major depression. Four focus groups followed by 10 in-depth qualitative interviews were conducted. Narratives were analyzed using the constant comparative method informed by grounded theory. Need for formal care was described in relation to acute somatic symptoms, functional impairment, and mood changes. Treatment initiation occurred through family members and primary care physicians who encouraged or inhibited help-seeking. Adherence to depression care focused on interpersonal aspects of care, evaluated symptom relief, and improved functioning. Help-seeking barriers included self-reliance, language barriers, stigma, competing health demands, and structural barriers. Findings from this study highlight potential points of intervention for developing culturally-appropriate collaborative care approaches for low-income Latinos with diabetes and major depression.


Acute lymphocytic leukemia with 9p anomalies. A report of four additional cases and review of the literature.
Childhood acute lymphocytic leukemia (ALL) with partial deletion of the short arm of chromosome 9 (9p-), particularly in the p21-22 region, associated with bulky disease, has been regarded as a possible subgroup of ALL. We have reviewed clinical and cytologic data in 128 cases of ALL (childhood and adult). Four of them had 9p anomalies. Two patients had a deletion in the 9p21 region associated with another deletion (9p13----pter) in one case and with t(1;19)(q21;p13) in the second patient. A third patient had a t(9;14)(p21;q12) balanced translocation associated with 14q22----qter deletion; the last patient showed a t(5;9)(p14;q21) unbalanced translocation also associated with 14q deletion. All four patients had lymphomatous ALL, but immunophenotype was non-T, in the four cases, (non-T, non-B in two patients and common ALL in the two remaining cases). Acute lymphocytic leukemia with 9p anomalies appears relatively frequently and is usually associated with poor prognostic features (i.e., bulk disease and high leukocyte counts) but does not seem restricted to childhood and T-cell lineage.


Infant dreaming and fetal memory: a possible explanation of sudden infant death syndrome.
During rapid-eye-movement sleep, when we dream, the brain is thought to be processing stored memory. The memory of a newborn infant is dominated by its fetal experience, and the infant is likely to dream about its life in the womb. Research with lucid (or conscious) dreaming has shown that dream images are supported by the corresponding body actions, using those muscles which remain active during rapid-eye-movement sleep. We suggest that sudden infant death syndrome or cot death may be a result of an infant dreaming about its life (or memory) as a fetus. In the course of that dream, since a fetus does not breathe (in the usual sense) the infant may cease to breathe and may die. This simple hypothesis is consistent with all of the known facts about sudden infant death syndrome (pathological and epidemiological), such as the age at death curve (the observed exponential decay and possibly the peak at 2-3 months), the higher risk with the prone sleeping position (but not excluding the supine position), and the observed climatic variation (seasonal and regional) in the incidence of sudden infant death syndrome. Many of these well-established facts have no other known explanation and other theories can generally only account for a few of the known facts about sudden infant death syndrome. Our hypothesis is also supported by recent findings that, as a group, sudden infant death syndrome infants have a higher proportion of rapid-eye-movement sleep, and also that they have an average higher heart rate (corresponding to possible fetal dreams) but only during rapid-eye-movement sleep.(ABSTRACT TRUNCATED AT 250 WORDS)


Prevalence and reasons influenced medication non-adherence among diabetes patients: A mixed-method study.
This study aimed to identify the prevalence of medication non-adherence (MNA) and to explore the reasons that influenced MNA among diabetes patients. This study used the explanatory mixed-method design. Phase one comprised of a cross-sectional study followed by phase two of a qualitative study. This study took place at two public hospitals in the Klang Valley, Malaysia. About 427 diabetes patients were recruited and 399 of them completed the study. The inclusion criteria were those with age more than 18 years and above, Malaysian citizen, able to understand Malay or English, and were diagnosed with diabetes mellitus for more than one year. The exclusion criteria were those with an intellectual disability and pregnant women. Phase two involved 12 participants recruited from non-adherent patients in phase one of the study. About 46.6% of the patients failed to adhere to the medication. Malays (OR: 1.66, 95%CI: 1.09 to 2.51, p = 0.017), single/widow or divorced (OR: 1.79, 95%CI: 1.05 to 3.05, p = 0.031) and poor HbA1c (OR: 2.57, 95% CI: 1.61 to 4.10, p = < 0.01) were associated with medication non-adherence. Five main categories emerged as the reasons for medication non-adherence, including perceived benefit of Complementary and Alternative medicine, attitude towards drawback of western medication, poor healthcare providers and patients' relationship, undesirable emotional response towards medication intake, as well as restraints in daily routine and cognitive function. There are many reasons for patients' non-adherence to their anti-diabetes medication. These findings are important in identifying the factors that influenced non-adherence to recommend reliable patient-centred care strategies in improving medication non-adherence among patients with diabetes.


Analytical methods for the characterization of surface finishing in bricks.
The recent restoration works of Santo Stefano Church Façade (XV century) in Venice have shown traces variously saved of different kind of surface finishes. These finishes were found on the brick's surface both in the masonry and in the decorative elements. Different brick's surface and decorative tile samples were investigated using several techniques: optical microscopy, scanning electron-microscopy, thermal analysis, infrared spectroscopy and reflectance Fourier transform infrared microspectroscopy. The evaluation of the reached results was used to understand the decorative techniques and to recognize the material employed.


Improved Ruthenium II tris (bathophenantroline disulfonate) staining and destaining protocol for a better signal-to-background ratio and improved baseline resolution.
In proteomics the ability to visualize proteins from electropherograms is essential. Here a new protocol for staining and destaining gels treated with Ruthenium II tris (bathophenantroline disulfonate) is presented. The method is compared with the silver-staining procedure of Swain and Ross, the Ruthenium II tris (bathophenantroline disulfonate) stain described by Rabilloud (Rabilloud T., Strub, S. M. Luche, S., Girardet, S. L. et al., Proteomics 2001, 1, 699-704) and the SYPRO Ruby gel stain. The method offers a better signal-to-background ratio with improved baseline resolution for both sodium dodecyl sulfate-polyacrylamide gels and two-dimensional gels.


Comparison of seroma production in breast conserving surgery with or without intraoperative radiotherapy as tumour bed boost.
One of the most common complications in breast conserving surgery is seroma formation. The origin of seroma formation remains unclear. While intraoperative radiotherapy (IORT) has been shown to be an alternative to whole breast irradiation, the influence on seroma production is unclear. Therefore, this analysis compares seroma production in patients with breast conserving surgery with or without IORT as tumour bed boost during breast conserving surgery. A retrospective analysis of seroma production in patients with nodal-negative (pN0sn) pT1/2 primary breast cancer treated between September 2010 and October 2013 at the Breast Cancer Centre, University Hospital Ulm was performed. Patients with neoadjuvant chemotherapy, previous breast/axillary surgery or more than one intervention were excluded. IORT was applied as a tumour bed boost with 50-kV X-rays (Intra beam(®)) delivering 9 Gy at the applicator surface. Seroma formation was measured using wound drains placed in breast and in axilla. Data of 152 patients (99 -IORT; 53 +IORT) were available for analysis. No significant differences between patients with or without IORT with regard to seroma production and number of days until drain removal were found (all p > 0.05). Patients with IORT encountered no increased seroma production and removal of the drains was not delayed compared to patients with breast conserving surgery only. Our results indicate that IORT does not increase the seroma production compared to surgery alone.


Global status of visceral leishmanial infection among blood donors: A systematic review and meta-analysis.
Transmission of Leishmania through transfusion has been reported from various Visceral leishmaniasis (VL) endemic areas of the world. The true burden of Leishmania infection in blood donors remains generally unknown. Thus, the present systematic review attempted to determine the global prevalence of Leishmania infection among blood donors. Data were extracted through five English and five Persian databases during the period from 1997 to 2016. Overall, 16 articles fulfilled the inclusion criteria and were used for data extraction in this systematic review. In total, 13,743 blood donors from different regions of world were examined. The prevalence rate of Leishmania infection according to seropositivity obtained 7% (95%CI: 5%, 8%). The lowest and the highest prevalence were related to Bangladesh 0.25% (95%CI: 0.0%, 1.0%) and Brazil, 16% (95%CI: 12%, 19%). Seroprevalence rate of leishmaniasis among females was more (4.60%) than males. Of 15 studies included in the meta-analysis, the pooled prevalence rate of molecular tests was obtained 2% (95%CI: 1%, 3%) in which Iran and Spain had the lowest and the highest prevalence, 0.05% and 7%, respectively. Our analysis showed that L. infantum was more common than L. donovani as etiological agent of VL among all donors. Our data confirms the presence of asymptomatic carriers of VL in endemic areas and supplies as an attentive to the likelihood of these carriers acting as blood donors. Moreover, we conclude that molecular tests for screening in asymptomatic blood donor provide an accurate estimate of the rate of infection over serological tests.


Cellular migration associated with macular hole: a new method for comprehensive bird's-eye analysis of the internal limiting membrane.
To elucidate the pathogenesis of macular hole formation, focusing in particular on the possible role of cellular migration on the cortical vitreous and internal limiting membrane (ILM) around the macular hole. To gain a comprehensive overview of the ILM excised in macular hole surgery (n = 36), the ILMs were carefully unfolded and spread out onto glass slides as continuous flat sheets that each contained a macular hole. The specimens were observed by light microscopy and transmission electron microscopy (n = 9), and the cellular distribution was analyzed by scanning electron microscopy in a quantitative manner (n = 27). Immunohistochemistry for glial fibrillary acidic protein and cytokeratin 18 was carried out for cellular characterization. Cellular proliferation was assessed by immunohistochemistry for proliferating cell nuclear antigen and Ki-67. Cellular migration was not apparent around the macular hole in the early stage of development of the macular hole (stage 2, 0 microm). As the macular hole passed through the later stages of development, cellular migration developed around the macular hole (stage 3, 84 microm) and the area of cellular migration gradually enlarged (stage 4, 420 microm). The immunophenotypic analysis showed that these cells were mainly glial fibrillary acidic protein-positive glial cells and cytokeratin 18-positive retinal pigment epithelial cells. The proliferating cell nuclear antigen and Ki-67 immunohistochemistry showed that some of these cells were proliferating on the ILM. Cellular migration on the ILM is not necessary for the initial formation of a macular break. Cellular migration developed after the macular break occurred, and the migration and proliferation increased gradually from the macular hole. This study provides a new method for understanding the ultrastructural analysis of the pathogenesis of the macular hole.


Circadian profiles of focal epileptic seizures: a need for reappraisal.
Circadian rhythm of seizure is underestimated in the study of focal epilepsies. A review of the current literature revealed a clear correlation between cortical epileptogenic focus and the circadian phase of seizure peak occurrence in adult patients. A single diurnal peak at 19:00 was found in seizures originating from the occipital lobe, between 5:00 and 7:00 in frontal lobe seizures, and between 16:00 and 17:00 h in temporal lobe seizures. Two diurnal peaks, between 5:00 and 7:00, and at 23:00 are reported in seizures from the parietal lobe, and between 7:00 to 8:00 and 16:00 to 17:00 in mesial temporal onset seizures. This circadian character of seizure occurrence in focal epilepsies may not be unique to partial seizures since recent clinical and experimental data indicate that generalized seizures also demonstrate circadian effects. The clinical evidence on generalized seizures and epilepsies is not recent, but a formal integration of circadian rhythmicity in our understanding and clinical management of epilepsies may be warranted.


Pedunculopontine nucleus microelectrode recordings in movement disorder patients.
The pedunculopontine nucleus (PPN) lies within the brainstem reticular formation and is involved in the motor control of gait and posture. Interest has focused recently on the PPN as a target for implantation of chronic deep brain stimulation (DBS) electrodes for Parkinson's disease (PD) and progressive supranuclear palsy (PSP) therapy. The aim of this study was to examine the neurophysiology of the human PPN region and to identify neurophysiological landmarks that may aid the proper placement of DBS electrodes in the nucleus for the treatment of PD and PSP. Neuronal firing and local field potentials were recorded simultaneously from two independently driven microelectrodes during stereotactic neurosurgery for implantation of a unilateral DBS electrode in the PPN in five PD patients and two PSP patients. Within the PPN region, the majority (57%) of the neurons fired randomly while about 21% of the neurons exhibited 'bursty' firing. In addition, 21% of the neurons had a long action potential duration and significantly lower firing rate suggesting they were cholinergic neurons. A change in firing rate produced by passive and/or active contralateral limb movement was observed in 38% of the neurons that were tested in the PPN region. Interestingly, oscillatory local field potential activity in the beta frequency range ( approximately 25 Hz) was also observed in the PPN region. These electrophysiological characteristics of the PPN region provide further support for the proposed role of this region in motor control. It remains to be seen to what extent the physiological characteristics of the neurons and the stimulation-evoked effects will permit reliable identification of PPN and determination of the optimal target for DBS therapy.


The central site of the sympatho-inhibitory action of 5-hydroxytryptamine in the cat.
The aim of this study was to determine the site in the CNS at which 5-hydroxytryptamine (5-HT) inhibits efferent sympathetic nerve activity in the cat. 5-Hydroxytryptamine (3 and 10 micrograms/kg), given into the lateral cerebral ventricle, produced immediate non dose related increases in mean blood pressure (MBP), heart rate (HR) and renal nerve activity (RNA). Larger doses (30 and 100 micrograms/kg i.c.v.) produced gradual decreases in blood pressure, heart rate and renal nerve activity, which did not occur when access of the drug to the fourth ventricle was prevented. Administration of 5-HT (10 and 30 micrograms/kg) into the fourth ventricle produced only decreases in blood pressure, heart rate and renal nerve activity after 15-40 min, which were accompanied by decreases in cardiac output and renal vascular resistance, but little or no change in total peripheral resistance. Application of 5-HT onto the ventral surface of the medulla, into the subarachnoid space at various levels along the spinal cord or into various parts of the nucleus tractus solitarius produced no effect on blood pressure heart rate or renal nerve activity. However, application of a cotton wool pledget soaked in a 5-HT solution (3 mg/ml) over the entire obex/NTS region produced immediate decreases in blood pressure, heart rate and renal nerve activity. These studies suggest that the sympatho-inhibitory effect of 5-HT is due to an action at a site near the caudal end of the dorsal surface of the medulla.


Novel prostate cancer biomarkers derived from autoantibody signatures.
Due to the low specificity of the prostate-specific antigen (PSA) assay and a high false positive rate, a large number of prostate cancer (PCA) biopsies are performed unnecessarily. Consequently, there is a need for new biomarkers that can identify PCA at any stage of progression while limiting the number of false positives. The use of autoantibody signature-developed biomarkers has proven to be an effective method to solve this problem. Using T7 phage-peptide detection, we identified a panel of eight biomarkers for PCA on a training set. The estimated receiver-operating characteristic (ROC) curve had an area under the ROC curve of 0.69 when applied to the validation set. Spearman correlations were high, within 0.7 to 0.9, indicating that the biomarkers have a degree of inter-relatedness. The identified biomarkers play a role in processes such as androgen response regulation and cellular structural integrity and are proteins that are thought to play a role in prostate tumorigenesis. Autoantibodies against PCA can be developed as biomarkers for detecting PCA. The scores from the algorithm developed here can be used to indicate a relative high or low risk of PCA, particularly for patients with intermediate (4.0 to 10 ng/ml) PSA levels. Since most commercially available assays test for PSA or have a PSA component, this novel approach has the potential to improve diagnosis of PCA using a biologic measure independent of PSA.


Ishihara color plates as a test for simultanagnosia.
To alert ophthalmologists to the use of Ishihara color plates in the detection of simultanagnosia. We examined seven patients referred for impaired vision. Evaluation included color plate testing with Ishihara color plates. All seven patients had simultanagnosia, with marked difficulty in identifying the numbers in Ishihara color plates despite adequate visual acuity and the ability to name all of the colors in the plates correctly. One of these patients was referred with the diagnosis of a cone dystrophy because of her poor performance on the Ishihara test. All of the patients had bilateral occipitoparietal damage or atrophy on magnetic resonance imaging. Ophthalmologists must be aware that a poor performance with Ishihara plates may not be attributable to an impairment of color vision but rather to occipitoparietal brain damage associated with simultanagnosia.


New theory of hippocampal function: associated rehearsal of multiplexed coded symbols.
A new theory of the role of the hippocampus in the selective storage of information in long-term memory is presented. This theory is based on the very recent discovery that neurons in the mammalian cerebral cortex transmit extremely precise copies of patterns of discharge in time when specific sensory inputs are presented, patterns that are interpreted to code for or symbolize specific items of information. The theory incorporates and provides an explanation for both the complex and unique internal structure of the hippocampal formation (the hippocampus and associated dentate gyrus) and the roles of many of the direct and indirect connections that structure makes with other brain structures. It also explains the deficits in learning that result from damage to the hippocampus and/or tracts that provide inputs to (or outputs from) this body as well as the role of the hippocampal formation in mapping the relationship of an individual to objects in its environment. The proposed enplanation is as follows. The hippocampal formation functions as a coordinated structure that, specifically, generates multiple copies of two different kinds of symbols (i.e., specific patterns of trains of nerve discharges in time). These two kinds of patterns are respectively provided by the entorhinal cortex through the perforant-alvear pathways and by the septal region, through the fornix, one of the two inputs to and the only output from the hippocampal formation. These two separate and different kinds of patterns are used to make multiplexed patterns that are ultimately transmitted to the cingulate gyrus and from there to other cortical memory storage locations. This transmission of amplified representations of different symbols occurs through the fornix. From there they are either transmitted to the mammilary bodies in the hypothalamus and from there to the anterior thalamic nuclei or, alternatively, directly to the anterior thalamic nuclei, bypassing the mammilary bodies. These thalamic nuclei in turn project the information to the cingulate gyrus of the cortex. The effect of the transmission of these mixtures of symbols is to cause the coordinated rehearsal and selective storage of relationships between separate inputs (specifically, patterns of discharge that symbolize different aspects of input) that are of probable significance to the survival of the system. The repeated presentation of such specific combinations of representations (symbols) then causes rehearsal-consolidation of these symbol associations as more permanent memories.(ABSTRACT TRUNCATED AT 400 WORDS)


Squamous cell carcinoma presenting as a painful scar following a single injury.
We report a case of squamous cell carcinoma of the skin presenting as a painful scar, following a single injury. There was no ulceration of the overlying tissues. The possible pathogenesis, with reference to mechanical and chemical trauma, is discussed.


[Histological and electron microscopic study of the cerebral cortex of the progeny of neurosensitized rats].
The sensomotor brain cortex of 30-day ratlings born by preliminary neurosensitized females, was studied. The studies demonstrated retardation in the development of the brain cortex in the progeny. Electon microscopic examinations detected certain features in the structure of cellular organella, as well as membrane formations in the nuclei of the cortical neurons. These changes are considered as an expression of a lowered level of neuron functioning.


Differential activation of LFA-1 and Mac-1 ligand binding domains.
Nine integrin alpha subunits contain an 'inserted' or I-domain, known to involve in ligand binding. Mutation of an invariant isoleucine residue in the I-domains of alphaL and alphaM has previously been reported to activate LFA-1 and Mac-1, respectively. In this article, we report notable differences in the regulation of adhesion of these two integrins. We find that mutation of the isoleucine residue in the proposed "socket for isoleucine" in full-length alphaL does not lead to an active LFA-1, although mutation of the equivalent residue in alphaM does convey constitutive activity to Mac-1. In addition, we observe the isolated I-domain of alphaL to be constitutively active. This challenges reports that state the alphaL I-domain exists in an inactive, closed conformation, and requires the presence of activating agents for ligand binding. These results shed further light on the many questions surrounding regulation of integrin activation.


Considerations for observational study design: Comparing the evidence of opioid use between electronic health records and insurance claims.
Pharmacoepidemiology studies often use insurance claims and/or electronic health records (EHR) to capture information about medication exposure. The choice between these data sources has important implications. We linked EHR from a large academic health system (2015-2017) to Medicare insurance claims for patients undergoing surgery. Drug utilization was characterized based on medication order dates in the EHR, and prescription fill dates in Medicare claims. We compared opioid use documented in EHR orders to prescription claims in four time periods: 1) Baseline (182 days before surgery); 2) Perioperative period; 3) Discharge date; 4) Follow-up (90 days after surgery). We identified 11 128 patients undergoing surgery. During baseline, 34.4% (EHR) versus 44.1% (claims) had evidence of opioid use, and 56.9% of all baseline use was reflected only in one data source. During the perioperative period, 78.8% (EHR) versus 47.6% (claims) had evidence of use. On the day of discharge, 59.6% (EHR) versus 45.5% (claims) had evidence of use, and 51.8% of all discharge use was reflected only in one data source. During follow-up, 4.3% (EHR) versus 10.4% (claims) were identified with prolonged opioid use following surgery with 81.4% of all prolonged use reflected only in one data source. When characterizing opioid exposure, we found substantial discrepancies between EHR medication orders and prescription claims data. In all time periods assessed, most patients' use was reflected only in the EHR, or only in the claims, not both. The potential for misclassification of drug utilization must be evaluated carefully, and choice of data source may have large impacts on key study design elements.


Optic Nerve Crush Injury in Rodents to Study Retinal Ganglion Cell Neuroprotection and Regeneration.
Optic nerve crush injury is a useful model for studying the response of central nervous system neurons (CNS) to injury. A particular focus of this model has been to elucidate therapeutic factors in promoting neuroprotection and axon regeneration after injury. Here we describe a step-by-step protocol in accessing the optic nerve and creating a crush injury. This can be used to create a reproducible model to study the response of retinal ganglion cells (RGC), the main projection neurons of the eye, to injury.


Proteomic analysis reveals mechanisms underlying increased efficacy of bleomycin by photochemical internalization in bladder cancer cells.
Photochemical internalization (PCI) is a promising new technology for site-specific drug delivery, developed from photodynamic therapy (PDT). In PCI, light-induced activation of a photosensitizer trapped inside endosomes together with e.g. chemotherapeutics, nucleic acids or immunotoxins, allows cytosolic delivery and enhanced local therapeutic effect. Here we have evaluated the photosensitizer meso-tetraphenyl chlorine disulphonate (TPCS2a/fimaporfin) in a proteome analysis of AY-27 rat bladder cancer cells in combination with the chemotherapeutic drug bleomycin (BML). We find that BLMPCI attenuates oxidative stress responses induced by BLM alone, while concomitantly increasing transcriptional repression and DNA damage responses. BLMPCI also mediates downregulation of bleomycin hydrolase (Blmh), which is responsible for cellular degradation of BLM, as well as several factors known to be involved in fibrotic responses. PCI-mediated delivery might thus allow reduced dosage of BLM and alleviate unwanted side effects from treatment, including pulmonary fibrosis.


[Biocompatibility of combined deproteinized bone coated with hepatocyte growth factor as scaffold for osteoblasts in vitro in fetal rabbits].
To determine the cellular compatibility of combined deproteinized bone(DPB) coated with hepatocyte growth factor (HGF), and to observe the adherent effect of osteoblasts in response to HGF. Osteoblasts were isolated from fetal rabbits. Osteoblasts were cultured with DPB coated with HGF and deproteinized bone as experimental group and contral group, respectively. The proliferation and alkalinephosphatase activity were tested. Their growth was examined by inverted phase contrast microscope and scanning electronmicroscope. The osteoblasts were attached to the outside and inside surfaces and grew well. HGF/DPB could stimulate the alkalinephosphatase activity of the osteoblasts and improve the proliferation of the osteoblasts. HGF/DPB has good biocompatibility and bone induction. HGF could improve the adherent effect of DPB on osteoblasts, and it could be used as scaffold material for the bone tissue engineering.


Multicenter clinical and functional evidence reclassifies a recurrent noncanonical filamin C splice-altering variant.
Truncating variants in filamin C (FLNC) can cause arrhythmogenic cardiomyopathy (ACM) through haploinsufficiency. Noncanonical splice-altering variants may contribute to this phenotype. The purpose of this study was to investigate the clinical and functional consequences of a recurrent FLNC intronic variant of uncertain significance (VUS), c.970-4A>G. Clinical data in 9 variant heterozygotes from 4 kindreds were obtained from 5 tertiary health care centers. We used in silico predictors and functional studies with peripheral blood and patient-specific induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). Isolated RNA was studied by reverse transcription polymerase chain reaction. iPSC-CMs were further characterized at baseline and after nonsense-mediated decay (NMD) inhibition, using quantitative polymerase chain reaction (qPCR), RNA-sequencing, and cellular electrophysiology. American College of Medical Genetics and Genomics (ACMG) criteria were used to adjudicate variant pathogenicity. Variant heterozygotes displayed a spectrum of disease phenotypes, spanning from mild ventricular dysfunction with palpitations to severe ventricular arrhythmias requiring device shocks or progressive cardiomyopathy requiring heart transplantation. Consistent with in silico predictors, the c.970-4A>G FLNC variant activated a cryptic splice acceptor site, introducing a 3-bp insertion containing a premature termination codon. NMD inhibition upregulated aberrantly spliced transcripts by qPCR and RNA-sequencing. Patch clamp studies revealed irregular spontaneous action potentials, increased action potential duration, and increased sodium late current in proband-derived iPSC-CMs. These findings fulfilled multiple ACMG criteria for pathogenicity. Clinical, in silico, and functional evidence support the prediction that the intronic c.970-4A>G VUS disrupts splicing and drives ACM, enabling reclassification from VUS to pathogenic.


Progression of the renal graft: treatment of acute rejection based on a biopsy against a presumptive diagnosis.
The aim of this study was to compare the progression of renal grafts following treatment of an acute rejection event based on the histological diagnosis of a graft biopsy compared to a presumptive (clinical and laboratory) diagnosis. A historical cohort was used to study 44 patients undergoing a living haploidentical related donor renal transplant, using a similar immunosuppressive treatment: cyclosporine, azathioprine, and prednisone. Acute rejection events were treated with methylprednisolone (250 mg for 3 to 5 days) based on a histological diagnosis (biopsy group = 14) or on a clinical and laboratory diagnosis (presumptive group = 30), which consisted of an elevation over 20% in plasma creatinine in 24 hours and renal ultrasound or scintigraphy findings. The study demonstrated no significant difference in renal function (plasma creatinine) and other outcomes 2 years following transplantation in both groups. The results show that treatment of acute rejection based on a presumptive diagnosis is not a risk factor for unfavorable outcomes following 2 years of renal transplantation monitoring.


Bid is involved in apoptosis induced by Chinese giant salamander iridovirus and contributes to the viral replication in an amphibian cell line.
Bid is a pro-apoptotic BH3-only member of the Bcl-2 superfamily that functions to link the extrinsic apoptotic pathway and the mitochondrial amplification loop of the intrinsic pathway. In this study, the expression and functions of Chinese giant salamander (Andrias davidianus) Bid (AdBid) were investigated. The AdBid cDNA sequence contains an open reading frame (ORF) of 576 nucleotides, encoding a putative protein of 191 aa. AdBid possesses the conserved BH3 interacting domain and shared 34-52% sequence identities with other amphibian Bid. mRNA expression of AdBid was most abundant in muscle. The expression level of AdBid in Chinese giant salamander muscle, kidney and spleen significantly increased after Chinese giant salamander iridovirus (GSIV) infection. Additionally, a plasmid expressing AdBid was constructed and transfected into the Chinese giant salamander muscle cell line (GSM cells). The morphology and cytopathic effect (CPE) and apoptotic process in AdBid over-expressed GSM cells was significantly enhanced during GSIV infection compared with that in control cells. Moreover, a higher level of the virus major capsid protein (MCP) gene copies and protein synthesis was confirmed in the AdBid over-expressed cells. These results indicated that AdBid played a positive role in GSIV induced apoptosis and the viral replication. This study may contribute to the better understanding on the infection mechanism of iridovirus-induced apoptosis.


Phenoloxidases in ascidian hemocytes: characterization of the pro-phenoloxidase activating system.
The phenoloxidase (PO) activity of the hemocytes lysate supernatant from three ascidians species, assayed by means of 3-methyl-2-benzothiazolinone hydrazone hydrochloride, have been compared. PO-containing hemocytes were identified by a cytochemical reaction and the enzymatic activity measured by a spectrophotometric assay of lysate supernatant from hemocyte populations separated on a discontinuous Percoll density gradient. In Styela plicata, the enzyme appeared to be contained in morula cells only. In Ciona intestinalis, PO activity was shown in univacuolar refractile granulocyte and granular hemocyte. In Phallusia mammillata both compartment cell and granular hemocytes were positive. Enzymatic assay following electrophoretic analysis on polyacrylamide gel electrophoresis (PAGE) or SDS-PAGE indicated that hemocyte lysate presented orthodiphenoloxidase (catecholase) activity. The enzymes from the three species differed in molecular size, activating substances and trypsin sensitivity.


Feed contaminated by fumonisin (Fusarium spp.) in chicks has a negative influence on oxidative stress and performance, and the inclusion of curcumin-loaded nanocapsules minimizes these effects.
The aim of this study was to determine whether the addition of curcumin (free and encapsulated) to chick feed would minimize the negative effects on health and performance caused by daily intake of fumonisin. We used 50 birds, divided into five treatments: CP, basal diet with 600 mg/kg of fumonisin, with antibiotic and coccidiostatic agent; CU, 600 mg/kg of fumonisin and 50 mg/kg of curcumin; NC5, feed with 600 mg/kg of fumonisin and 5 mg of nano-curcumin/kg of feed; NC10, feed with 600 mg/kg of fumonisin and 10 mg of nano-curcumin/kg of feed; and CN, fumonisin-free diet, with antibiotic and coccidiostatic. We measured weights, weight gain, and serum biochemistry, as well as antioxidant and oxidant activities. Lower body weight and weight gain were observed in chicks that received feed with fumonisin; curcumin did not minimize this negative effect. Lower glucose and triglyceride levels were also observed in the NC10 group, while the highest cholesterol levels were observed in all groups of birds that consumed fumonisin compared to the CN group. Uric acid levels were significantly lower in CP than in CN. Levels of liver enzymes were higher in CP than in CN. The highest levels of thiobarbituric acid reactive substances were found in CP and CU, whereas ROS was higher in CU compared to CN. Superoxide dismutase activity was significantly lower in CP, while glutathione S-transferase activity was higher in the CP group. Catalase activity was lower in groups of birds that consumed fumonisin compared to CN. Taken together, these findings suggest that intake of curcumin-loaded nanocapsules (10 mg/kg) had hepaprotective and antioxidant effects in chicks artificially intoxicated with fumonisin, minimizing the negative effects caused by this mycotoxin.


Haemorrhage-induced production of Fos in neurons of the lamina terminalis: role of endogenous angiotensin II.
Hypotensive haemorrhage increased the production of Fos, a marker of cell activation, in cells of the subfornical organ (SFO) and the organum vasculosum of the lamina terminalis (OVLT) as well as in the supraoptic (SON) and paraventricular (PVN) nuclei in conscious rats. Pretreatment with captopril (100 mg/kg i.p.) inhibited the production of Fos in cells of the SFO and OVLT but did not reduce the concentration of Fos-positive cell nuclei in the PVN and SON. The results suggest that the production of Fos in the SFO and OVLT induced by the haemorrhage is probably due to elevated levels of circulating angiotensin II.


Singing Voice Quality: The Effects of Maxillary Dental Arch and Singing Style.
In classical singing techniques, it is common to manipulate the vocal tract to channel airflow to increase voice quality and volume. Technique varies according to the style of the music, the voice type, and range of a given singer. Although these practices are intentional, fixed physiological aspects of a singer's vocal instrument also play an extremely impactful role in determining voice quality. In the present study, the relationship between the dimensions of the maxillary dental arch and voice quality were examined in professional singers. The dimensions of the palate were measured from the maxillary dental casts of 14 female singers. Audio recordings were made for the same participants while singing a sustained /a/ singing vowel, a glissando, the song "Are You Sleeping", and a selected song from their personal repertoire. The dimensions of the palate were measured from maxillary dental casts. From the recordings, two parameters were calculated: (1) the Singing Power Ratio (SPR) and (2) A2 A1 ratio. Higher SPR values indicate a stronger ring in the voice, typical of operatic singing style, while higher A2 A1 ratio values are associated with the belting singing style. Singers with larger frontal palate depth, smaller posterior palate depth, larger frontal palate width, and smaller posterior palate width seem to be more suitable for an operatic singing style. Singers who had larger overall depth and width of the palate measurements produced an increased second harmonic, typical of the belting style. When considering a singer's ability to produce vocalizations successfully, physiological structure is an increasingly important factor. The present study discovered that palate depth and width are associated with statistically significant differences in SPR and A2/A1 parameters. These parameters correlate with two styles of singing, operatic, and belting respectively.


Effects of pancreatectomy on nutritional state, pancreatic function and quality of life.
There are concerns about the extent of impaired endocrine and exocrine pancreatic function and poor quality of life (QoL) after pancreatectomy, but there is little information from large prospective follow-up studies. Consecutive patients undergoing pancreaticoduodenectomy or distal pancreatectomy between 2007 and 2011 were included. Relative bodyweight (RBW), triceps skinfold thickness (TSFT), serum protein, albumin, transferrin, fasting blood glucose, postprandial 2-h glucose (PP2), glycosylated haemoglobin A1c and stool elastase measurements, and European Organization for Research and Treatment of Cancer QLQ-C30 questionnaires were collected serially for 1 year. Some 136 patients undergoing pancreatic resection completed the study. RBW and TSFT recovered to over 90 per cent of the preoperative value by 12 months, whereas transferrin, albumin and protein had returned to preoperative levels by 3 months. Diabetes mellitus, impaired fasting glucose or raised PP2 was present in 42 of 76 patients at 6 months and 36 of 76 at 12 months. Although steatorrhoea and diarrhoea had mainly resolved by 3 months, stool elastase level decreased after operation and showed no recovery. Nutritional status, pancreatic endocrine function and QoL returned to preoperative levels in 63 (46·3 per cent), 72 (52·9 per cent) and 77 (56·6 per cent) of 136 patients within 6 months of pancreatectomy. Multivariable analysis revealed that age 60 years or more, operation type, chronic pancreatitis and malignant disease had a significant impact on nutritional index, pancreatic function and QoL. About half of all patients can expect recovery from pancreatectomy after 6 months, but those with risk factors need more careful follow-up and supportive management.


Psychrobacter piechaudii shunt infection: first report of human infection.
Psychrobacter piechaudii is a recently described species of Gram-negative bacteria in the Moraxellaceae family. No cases of human infection due to this species have been described before. We report the case of an ex-premature infant girl with hydrocephalus secondary to intraventricular haemorrhage who underwent multiple cerebrospinal fluid (CSF) shunt operations. She ultimately developed Psychrobacter piechaudii meningitis, presenting as ventriculoperitoneal shunt dysfunction and wound leak, which necessitated removal of the shunt, a period of external ventricular drainage and antibiotics. We found this organism to be sensitive to intravenous ceftazidime (50 mg/kg) and ciprofloxacin, and a 7-10 day treatment course prior to shunt re-insertion (and 3 week total course) was sufficient. The patient is well post-operatively. To the best of our knowledge, this is the first reported case of Psychrobacter piechaudii infection in a human.


Chronic neural activity recorded within breast tumors.
Nerve fibers are known to reside within malignant tumors and the greater the neuronal density the worse prognosis for the patient. Recent discoveries using tumor bearing animal models have eluded to the autonomic nervous system having a direct effect on tumor growth and metastasis. We report the first direct and chronic in vivo measurements of neural activity within tumors. Using a triple-negative mammary cancer mouse model and chronic neural interface techniques, we have recorded neural activity directly within the tumor mass while the tumor grows and metastasizes. The results indicate that there is a strong connection between the autonomic nervous system and the tumor and could help uncover the mechanisms of tumor growth and metastasis.


Inhibition of uterine contractility with various tocolytics with and without progesterone: in vitro studies.
Various tocolytics are used to suppress uterine contractility in patients in preterm labor. Progesterone (P4) is used in patients at high risk for preterm delivery. In this study, we evaluated the effects of various tocolytics with and without P4 to examine effects on uterine contractility. Uterine tissues (n = 280) from women undergoing cesarean at term were exposed in vitro to various agents (vehicle, magnesium sulfate [MgSO(4)], nifedipine, indomethacin, or pinacidil-all with and without P4). Contractility was measured before and after addition of the various agents. P4 alone at 10(-5) mol/L concentration has little effect to inhibit contractility (P ≥ .05). MgSO(4) (2-8 × 10(-3) mol/L) inhibits uterine contractility (P < .05) but there is no change when combined with P4 (P > .05). Nifedipine (10(-8) mol/L) and indomethacin (10(-5) mol/L) inhibit contractions alone (P < .05) and to a greater extent when combined with P4 (P < .05). P4 significantly (P < .05) reduced the effects of pinacidil (10(-6.5) mol/L). Combinations of P4 with nifedipine or indomethacin, but not MgSO(4), might be used to effectively suppress preterm labor.


Amyloid beta-protein aggregation nullifies its pathologic properties in cultured cerebrovascular smooth muscle cells.
Alzheimer's disease and related disorders are characterized by deposition of aggregated amyloid beta-protein (A beta) and accompanying pathologic changes in the neuropil and in the walls of cerebral blood vessels. A beta induces neurotoxicity in vitro, and this effect is markedly enhanced when the peptide is preaggregated. Recently, we reported that freshly solubilized A beta 1-42 can induce cellular degeneration and a striking increase in the levels of cellular amyloid beta-protein precursor and soluble A beta peptide in cultured cerebrovascular smooth muscle cells (Davis-Salinas, J., Saporito-Irwin, S. M., Cotman, C. W., and Van Nostrand, W. E. (1995) J. Neurochem. 65, 931-934). In the present study, we show that preaggregation of A beta 1-42 abolishes the ability of the peptide to induce these cellular pathologic responses in these cells in vitro. These findings suggest that distinct mechanisms for A beta-induced cytotoxicity exist for cultured neurons and cerebrovascular smooth muscle cells, supporting that different processes may be involved in the parenchymal and cerebrovascular pathology of Alzheimer's disease and related disorders.


Popcorn-shaped polyethylene synthesised using highly active supported permethylindenyl metallocene catalyst systems.
Unsymmetrical permethylindenyl bent metallocene complexes have been synthesised and reacted with inorganic solid supports to afford catalysts for the slurry phase polymerisation of ethylene. Those supported on solid polymethylaluminoxane were both highly active catalysts and afforded polymers with a desirable, low aggregation, "popcorn" morphology.


Novel stability of cellular neural networks with interval time-varying delay.
In this paper, the asymptotic stability is investigated for a class of cellular neural networks with interval time-varying delay (that is, 0<h1<dt<h2). By introducing a novel Lyapunov functional with the idea of partitioning the lower bound h1 of the time-varying delay, a new criterion of asymptotic stability is derived in terms of a linear matrix inequality (LMI), which can be efficiently solved via standard numerical software. The criterion proves to be less conservative than most of the existing results, and the conservatism could be notably reduced by thinning the delay partitioning. Two examples are provided to demonstrate the less conservatism and effectiveness of the proposed stability conditions.


A permanent plastic cannula for intraventricular injections into rat brain.
The construction and use of a permanent, implanted plastic camnula for intraventricular injections in rat brain is described. The cannula can be used in either acute or chronic experiments.


Thyrotoxicosis and hyperemesis gravidarum associated with a serum activity which stimulates human thyroid cells in vitro.
To investigate in-vitro thyroid stimulatory activity in the serum of patients with hyperemesis gravidarum and thyrotoxicosis. Serum from hyperthyroid patients was incubated with cultures of human thyroid cells. Attempts were made to neutralize stimulatory activity with antisera to hCG. Five patients presenting in early pregnancy with hyperemesis and thyrotoxicosis. Serum concentrations of thyroid hormones (total and free), TSH and hCG. Accumulation of extracellular cAMP in response to serum. All five patients had biochemical hyperthyroidism with no evidence of an underlying autoimmune disease. The mean cAMP accumulation over 4 hours with sera from 12 non-pregnant controls was 130.6 (121.8-142.8), from 12 pregnant controls 132.4 (118.1-143.8), compared with values of 144.7, 159.1, 166.2, 178.9 and 320.5 for the thyrotoxic patients. The stimulatory activity could not be neutralized by addition of anti-hCG or by depleting the sera of hCG. Thyrotoxicosis may present with hyperemesis in early pregnancy. Clinical and biochemical features may be masked by the pregnancy or by the intercurrent illness. The hypothesis that hCG is a thyroid stimulator in patients with hyperemesis gravidarum is not supported by the immuno-neutralization data in this study.


Effect of cold-induced pain on salt and water transport in the human jejunum.
The possibility that the central and autonomic nervous systems might be involved in the control of intestinal absorption was assessed by measuring the effect of physical stress (cold-induced pain) on jejunal transport in 13 healthy subjects. Using a triple-lumen perfusion technique to determine jejunal absorption, cold pain significantly reduced net water absorption from 34.9 to 15.7 ml/30 cm.50 min (p less than 0.005), net sodium absorption from 2.6 to 0.2 mEq/30 cm.50 min (p less than 0.005), and net chloride absorption from 2.2 to 0.6 mEq/30 cm.50 min (p less than 0.02). These changes were associated with an elevation of blood pressure and plasma norepinephrine during cold pain. During the period following cold pain, systolic blood pressure remained significantly elevated and net jejunal water absorption significantly reduced. These results suggest that physical stress can influence jejunal absorption of salt and water in humans and support the possibility that the central and autonomic nervous systems have a physiologic role in the control of intestinal function.


Structural basis of peptidomimetic agonism revealed by small- molecule GLP-1R agonists Boc5 and WB4-24.
Glucagon-like peptide-1 receptor (GLP-1R) agonists are effective in treating type 2 diabetes and obesity with proven cardiovascular benefits. However, most of these agonists are peptides and require subcutaneous injection except for orally available semaglutide. Boc5 was identified as the first orthosteric nonpeptidic agonist of GLP-1R that mimics a broad spectrum of bioactivities of GLP-1 in vitro and in vivo. Here, we report the cryoelectron microscopy structures of Boc5 and its analog WB4-24 in complex with the human GLP-1R and Gs protein. Bound to the extracellular domain, extracellular loop 2, and transmembrane (TM) helices 1, 2, 3, and 7, one arm of both compounds was inserted deeply into the bottom of the orthosteric binding pocket that is usually accessible by peptidic agonists, thereby partially overlapping with the residues A8 to D15 in GLP-1. The other three arms, meanwhile, extended to the TM1-TM7, TM1-TM2, and TM2-TM3 clefts, showing an interaction feature substantially similar to the previously known small-molecule agonist LY3502970. Such a unique binding mode creates a distinct conformation that confers both peptidomimetic agonism and biased signaling induced by nonpeptidic modulators at GLP-1R. Further, the conformational difference between Boc5 and WB4-24, two closed related compounds, provides a structural framework for fine-tuning of pharmacological efficacy in the development of future small-molecule therapeutics targeting GLP-1R.


Accuracy of contrast-enhanced voiding urosonography using Optison™ for diagnosis of vesicoureteral reflux in children.
There is limited quality of evidence regarding the accuracy of contrast-enhanced voiding urosonography (ceVUS) for diagnosis of vesicoureteral reflux (VUR) compared to fluoroscopic voiding cystourethrography (VCUG), and minimal data on the use of the ultrasound contrast agent Optison™ for this purpose. To compare the accuracy of ceVUS using Optison™ to VCUG, and to assess inter-rater agreement regarding presence and grading of VUR. In this retrospective investigation, all sequential ceVUS with Optison™ and VCUG studies performed in children between 2014 and 2017 were reviewed. Two raters independently graded all ceVUS studies using a 5-point scale. CeVUS sensitivity and specificity were estimated separately for each rater using the VCUG report as the ground truth for presence and degree of VUR. Logistic and ordinary linear regression models assessed rater-report agreement and inter-rater agreement for each kidney, Optison™ dose, and referral diagnosis. 97 children (51 females) with 101 paired studies were included. Sensitivity and specificity of ceVUS for VUR detection were identical for both raters: right kidney 75%/90.9%; left kidney 85.7%/78.9% (Figure). There was no statistically significant difference in disagreement between raters and the VCUG report for the right or left kidney. Inter-rater agreement on ceVUS grading was 90% and 88% for right and left kidneys, respectively. There was a significant negative association between fetal hydronephrosis vs urinary tract infection and disagreement between Rater 2 and the VCUG report for the left kidney. There were no other significant associations with respect to either kidney, Optison™ dose, or referral diagnosis. Our study showed that detection of VUR with ceVUS and Optison™ is comparable to fluoroscopic VCUG. Based on the VCUG reports, the incidence of VUR in our patient population was substantially lower than in the meta-analysis of Chua et al. and in the study of Kim et al. The explanation for the large discrepancy in VUR incidence may reflect differences in the patient populations, and in our reporting of VUR with respect to kidney number rather than to pelviureteral units. Study limitations include its retrospective nature and potential bias in terms of patient selection. Since VUR is an intermittent phenomenon, sequential rather than simultaneous performance of the ceVUS and fluoroscopic studies might have influenced VUR detection. A blinded comparison of ceVUS performed with Optison™ to fluoroscopic VCUG showed moderate-good sensitivity and specificity for diagnosis of VUR.