Comparison between aniridia with and without PAX6 mutations: clinical and molecular analysis in 14 Korean patients with aniridia.
To describe clinical and molecular characteristics of Korean patients with aniridia and to compare the clinical phenotype between those having an identifiable PAX6 mutation and those not. Comparative case series. A total of 14 Korean patients from 10 families with aniridia. Complete ophthalmologic examinations were performed for all patients. PAX6 analysis included direct sequencing of all coding regions and multiplex ligation-dependent probe amplification (MLPA) to detect large deletions when the sequencing was negative. If the PAX6 analysis failed to reveal any identifiable mutations, genomic copy number variation analysis via array comparative genomic hybridization (CGH) and candidate gene PITX3 and FOXE3 sequencing were then performed. Severity of ocular abnormalities and genetic findings. Sequencing of PAX6 exhibited 5 different heterozygous mutations in 8 patients from 5 families; 2 (p.Ser43Phe, IVS8-9C>G) were novel, and 3 (p.Arg208Trp, p.Arg317X, and p.X423L) have been previously reported. Among the remaining 6 patients in whom the PAX6 sequencing was negative, MLPA identified large deletions in 2 sporadic patients. However, the array CGH and candidate gene sequencing found no genomic or genetic abnormalities. The mutation detection rate was therefore 70%. Patients harboring an identifiable mutation in PAX6 had either a severe or a mild variant phenotype depending on the type of mutations. Likewise, among patients without an identifiable PAX6 mutation, their phenotypes varied widely from severe to very mild. This study adds 2 novel PAX6 mutations to those previously reported, providing further evidence for genetic and phenotypic heterogeneity in aniridic ocular malformation. There was no difference in the clinical phenotype between patients with and without detectable mutations in the PAX6 gene. The wide variability of ocular phenotype regardless of the presence or absence of PAX6 mutations calls for a further appreciation of the complexity in the molecular diagnosis of aniridia and suggests that this ocular malformation may be better regarded as a group of heterogeneous disorders, rather than a single disease entity, associated with mutations in PAX6 and/or other genes located elsewhere in the human genome. The authors have no proprietary or commercial interest in any of the materials discussed in this article.


A human monoclonal antibody reactive with human prostate.
A human immunoglobulin M monoclonal antibody secreting hybridoma, termed MHG7, has been isolated and characterized for its reactivity against human prostate cells. Lymphocytes isolated from a regional draining lymph node of a patient with prostate carcinoma were fused with murine P3-NS1-Ag4-1 myeloma cells. Supernatants from the generated mouse-human somatic cell hybrids were first screened for human immunoglobulin production by an enzyme immunoassay. The identified human immunoglobulin-secreting hybridomas were expanded for further analysis and their supernatants screened by enzyme immunoassay against a panel of prostate cell lines. The human immunoglobulin M monoclonal antibody MHG7, in addition to reacting with prostate cell lines, also reacted with prostate carcinoma cells and benign prostatic hypertrophy cells on both frozen and paraffin embedded tissue sections. These data suggest that regional draining lymph nodes of prostate carcinoma patients can be used as a source of human lymphocytes for generating human immunoglobulin-secreting hybridomas reactive with human prostate cells.


A protein from Ustilago which forms an acid-soluble complex with deoxyribonucleic acid.
A protein which can render DNA largely acid-soluble has been purified 1600-fold from high salt extracts of Ustilago maydis. The activity is unusual in that substrate DNA is not made acid-soluble through hydrolysis to small oligomers. Rather, the protein appears to bind to DNA to form a complex which itself is acid-soluble. The activity of conversion of DNA to an acid-soluble form is cold-labile, but the inactivation by cold is reversible by brief heat treatment. Divalent cations stimulate the activity; phosphate is inhibitory. Optimal activity is observed at pH 6.0 and again at pH 9.0. Nucleoside triphosphates and diphosphates stimulate activity at low protein concentrations but are not hydrolyzed during the course of reaction. The protein behaves anomalously on gel filtration columns and is completely excluded by Sephadex or agarose gels. When analyzed by sedimentation velocity, the protein was found to sediment at 5.3 S, the same rate at which a globular protein of 65,000 daltons would sediment. Dependence of activity upon protein concentrations is sigmoidal. K+ and to a lesser degree NH4+ are partially effective in abolishing the lag in the concentration curve. The protein displays a saturation curve when exposed to increasing DNA concentrations. Such a curve could only result from a non-random or cooperative mode of binding of the protein to DNA. A mutant sensitive to gamma and ultraviolet radiation with an abnormally high level of the protein has been found. Haploid populations of the mutant grow slowly and contain a large proportion (10 to 20%) of inviable cells. Diploids are defective in mitotic allelic recombination and fail to complete meiosis. It is speculated that the protein may be important in the regulation of chromosome condensation.


Synthesis, study on anti-arthritic, anti-inflammatory activity and toxicity of some novel bis-oxy cyclophane diamides.
A series of bis-oxy cyclophane diamides with bis(aminomethyl)m-terphenyl as spacer have been synthesized and characterized from spectral and analytical data. All the cyclophane diamides exhibit better anti-arthritic activity than the reference drug viz. diclofenac sodium. Some of the cyclophane diamides exhibit good anti-inflammatory activity. The cyclophane amide 4 and 5 do not show any evidence of mutagenicity and cytotoxicity.


Post-translational modification of Fibroblast Growth Factor 23.
Fibroblast Growth Factor (FGF) 23 has been shown to play important roles in the development of hypophosphatemic rickets/osteomalacia. Complementary DNA predicts that the FGF23 protein is composed of 251 amino acids and N-terminal 24 amino acids seem to be a signal peptide. In vitro experiments indicate that a part of the FGF23 protein is processed between arginine179 and serine180. When full-length, N-terminal and C-terminal processed fragments of FGF23 were injected into mice, only the full-length FGF23 reduced serum phosphate levels indicating that the processing of FGF23 abolished its effect to cause hypophosphatemia. This processing was shown to be prevented by an inhibitor of furin indicating that the cleavage is mediated by subtilisin-like proprotein convertase. In addition to this processing, FGF23 protein seems to have O-linked glycosylation. Further studies are necessary to clarify the importance of O-glycosylation for FGF23 activity.


Calibration of multi-layered probes with low/high magnetic moments.
We present a comprehensive method for visualisation and quantification of the magnetic stray field of magnetic force microscopy (MFM) probes, applied to the particular case of custom-made multi-layered probes with controllable high/low magnetic moment states. The probes consist of two decoupled magnetic layers separated by a non-magnetic interlayer, which results in four stable magnetic states: ±ferromagnetic (FM) and ±antiferromagnetic (A-FM). Direct visualisation of the stray field surrounding the probe apex using electron holography convincingly demonstrates a striking difference in the spatial distribution and strength of the magnetic flux in FM and A-FM states. In situ MFM studies of reference samples are used to determine the probe switching fields and spatial resolution. Furthermore, quantitative values of the probe magnetic moments are obtained by determining their real space tip transfer function (RSTTF). We also map the local Hall voltage in graphene Hall nanosensors induced by the probes in different states. The measured transport properties of nanosensors and RSTTF outcomes are introduced as an input in a numerical model of Hall devices to verify the probe magnetic moments. The modelling results fully match the experimental measurements, outlining an all-inclusive method for the calibration of complex magnetic probes with a controllable low/high magnetic moment.


Isolated Complete Heart Block in the Fetus.
Isolated congenital complete heart block (CCHB) is a rare disease with significant associated morbidity and mortality. A diagnosis is often made in fetal life, but data regarding long-term outcomes are limited, and fetal therapy to improve prognosis is controversial. In our institution, 85 fetuses were diagnosed with CCHB from 1981 to 2013 in 80 mothers. There were 37 anti-Ro-positive pregnancies, 36 both anti-Ro and anti-La positive, 10 antibody negative, and 2 of unknown antibody status. Antenatal treatments were given in 14 fetuses, with 8 given fluorinated steroids, 4 beta sympathomimetics, and both in 2. Of the original 85, 74 babies survived to delivery. Fetal hydrops was the only risk factor found to be significantly associated with intrauterine death (p <0.001). Four babies died before pacemaker implantation, 56 have had pacemakers implanted, and 14 are pacemaker free. The Kaplan-Meier estimate for median time to pacemaker implantation was 2.6 years, with 15 implanted in the neonatal period. There have been 14 postnatal deaths, with a Kaplan-Meier estimate of survival at 30 years of 76.8% (95% confidence interval 65% to 90%). Dilated cardiomyopathy was uncommon, occurring in 6 patients. Prematurity and hydrops were associated with increased postnatal mortality (p = 0.02 and 0.005, respectively). In conclusion, we present the largest single-unit experience of prenatally diagnosed CCHB in the published literature. Our cohort was conservatively managed, with survival similar to those previously published. These data offer insight into the long-term natural history of CCHB.


Abdominal pain with anorexia in patients with breast carcinoma.
Breast cancer is the second commonest primary tumour responsible for gastrointestinal metastases after malignant melanoma. The real incidence of gastrointestinal metastases in breast cancer patients is probably underestimated owing to the non-specific presenting symptoms and death of patients caused by other more obvious metastases. The predominant histological subtype of gastrointestinal metastases of breast cancer is invasive lobular carcinoma and the median interval from diagnosis of primary breast cancer to gastrointestinal metastases is five years. We report two cases of disseminated breast cancer with gastrointestinal involvement with a rather long survival.


Regulation of expression of the alternative mRNAs of the rat alpha-thyroid hormone receptor gene.
The rat alpha-thyroid hormone receptor gene encodes through alternative splicing at least three protein isoforms with different functions, and three mRNA species (2.6, 5.4, 6.8 kilobase (kb) in size) are detected using alpha gene-specific probes (Mitsuhashi, T., Tennyson, G. E., Nikodem, V. M. (1988) Proc. Natl. Acad. Sci. U. S. A. 85, 5804-5808). In the present study, the identities of these mRNAs were analyzed by Northern analysis, and it was demonstrated that in rat brain the receptor protein is encoded by the minor 5.4- and 6.8-kb mRNAs and the variant proteins are encoded by the major 2.6-kb mRNA. Relative quantities of these mRNAs were determined by RNase protection assay, and the ratio of the receptor mRNAs to the variant mRNAs was estimated to be 1:6 in adult brain. The ratio between the mRNAs was regulated in both a tissue-specific and developmental stage-specific manner. The receptor mRNA levels were also regulated by the thyroid state of the animal showing an increased level in hypothyroid rat liver while those in brain were not affected. Analysis of the alpha-thyroid hormone receptor gene suggested that the choice between two poly-adenylation sites and subsequent RNA processing appear to generate the 3' heterogeneity of these alternative mRNAs.


Detection of Malachite Green in Water Using Edge Excited Label Free Fluorescent Probe NCQDs.
The fluorescent properties of nitrogen doped carbon quantum dots (NCQDs) prepared through microwave assisted green method has been used as label free fluorescent probe for selective and sensitive detection of malachite green (MG) in water. The optical responses revealed that the NCQDs are highly stable and have good fluorescent quantum yield. The NCQDs were used to detect the Malchite Green in Mili Q water. Reduction in the fluorescence response was monitored in the range 17.12-128.43 μM of MG dissolved in Mili Q water. Linear response was observed in the range, 10-80 μM. The calculated value of limit of detection is 5.16 μM and the sensitivity is (0.03536 ± 0.00001) μM-1. The future application of this work is that it can be employed to detect MG in the tap water and other natural sources of water.


Role of the ER stress in prostaglandin E2/E-prostanoid 2 receptor involved TGF-β1-induced mice mesangial cell injury.
This study aims to investigate the relationship between prostaglandin E2 E-prostanoid 2 receptor (EP2) and Endoplasmic reticulum (ER) stress in transforming growth factor-β1 (TGF-β1)-induced mouse glomerular mesangial cells (MCs) injury. We cultured primary WT, EP2(-/-) MCs (EP2 deleted), and adenovirus-EP2-infected WT MCs (EP2 overexpressed). PCR, Western blot, flow cytometry, and immunohistochemical technique were used in in vitro and in vivo experiments. We found that TGF-β1-induced PGE2 synthesis decreased in EP2-deleted MCs and increased in EP2-overexpressed MCs. EP2 deficiency in these MCs augmented the coupling of TGF-β1 to ER stress-associated proteins [chaperone glucose-regulated protein 78 (GRP78), transient receptor potential channel 1 (TRPC1), and transient receptor potential channel 4 (TRPC4)], and upregulation of EP2 showed no significant change of GRP78, but augmented the expression of TRPC1, while TRPC4 expression was downregulated in comparison to normal MCs. In addition, EP2 deficiency in MCs augmented TGF-β1-induced fibronectin (FN), cyclooxygenase-2 (COX2), and CyclinD1 expression. Silencing of EP2 also strengthened TGF-β1-induced extracellular-signal-regulated kinase 1/2 (ERK1/2) phosphorylation. Flow Cytometry showed that silencing of EP2 significantly promoted the apoptosis of MCs. In contrast, EP2 overexpression reversed the effects of EP2 deficiency. 8 weeks after 5/6 nephrectomy (Nx), blood urea nitrogen and creatinine concentrations were significantly increased in EP2(-/-) 5/6Nx mice as compared to those of WT 5/6Nx mice. The pathological changes in kidney of EP2(-/-) mice were markedly aggravated compared with WT mice. Immunohistochemical analysis showed significant augment of TRPC4 and ORP150 in the kidney of EP2(-/-) mice compared with WT mice. Considering all the findings, it is suggested that increased expression of EP2 may prevent TGF-β1-induced MCs damage through ER stress regulatory pathway.


Inhibition of liver microsomal carnitine acyltransferases by sulphonylurea drugs.
The sulphonylureas glibenclamide and tolbutamide inhibited carnitine acyltransferase activities in rat liver microsomes. Glibenclamide was a more potent inhibitor than tolbutamide. The effect of tolbutamide on the malonyl-CoA-inhibitable transferase was influenced by the phospholipid/detergent environment whereas the effect of glibenclamide was not. Glibenclamide was a more potent inhibitor of the malonyl-CoA-inhibitable transferase than of the malonyl-CoA-insensitive enzyme. The extent of inhibition of the malonyl-CoA-inhibitable transferase by tolbutamide was similar to its effect on VLDL triacylglycerol secretion as reported by Wiggins and Gibbons [Biochem. J. 284 (1992) 457-462] possibly supporting the suggestion that microsomal carnitine acyltransferases are involved in VLDL triacylglycerol assembly/secretion.


Studies of cachexia in parasitic infection.
Cachexia and septic shock, syndromes associated with chronic and acute infection, respectively, are mediated by endogenous factors. The search for humoral mediators of cachexia led to the isolation of the cytokine known as cachectin/TNF. A biologic role for cachectin/TNF in cachexia has been identified: it induces a catabolic state in adipose and skeletal muscle cells in vitro, and animals persistently exposed to it in vivo become cachectic. Cachectin/TNF was also discovered to be a primary, essential mediator of septic shock. The acute administration of the recombinant protein causes shock, tissue injury, and derangements of metabolic homeostasis that mimic endotoxemia and septic shock syndrome. Anti-cachectin antibodies protect against the lethal effects of septic shock and cachexia, suggesting that this mediator has a central role as a trigger to the inflammatory and deleterious host responses to infection. Future investigations will undoubtedly be directed towards the effects of inhibiting cachectin/TNF in the treatment of the complications of infectious disease.


Use of capnography in diagnosis of pulmonary embolism during acute respiratory failure of chronic obstructive pulmonary disease.
In chronic obstructive pulmonary disease (COPD) patients, there is a difference between PaCO2 and end-tidal partial pressure of CO2 (PetCO2). This gradient P(a-et)CO2 is due to ventilation/perfusion mismatching and deadspace, and is usually abolished by forced and prolonged expiration. We hypothesized that this gradient might not be canceled by forced expiration in the case of acute respiratory failure (ARF) related to pulmonary embolism (PE). Forty-four adult COPD patients were prospectively entered into this study; they were suspected of having ARF related to PE on the basis of clinical and biological data on admission. Maximum expired partial pressure of CO2 (PemCO2) was measured in mechanically ventilated and sedated patients by an interrupt of mechanical support. CO2 concentration was recorded during the following prolonged and passive expiration. The test was considered valid if an expiratory plateau was obtained. PemCO2 was measured in triplicate. Simultaneously, PaCO2 was measured and the ratio, R = [( 1-PemCO2]/PaCO2) x 100, was calculated. Pulmonary angiography was performed on the same day for all patients. Results showed that 17 patients had PE (PE+) and 17 had no PE (PE-). The two groups were comparable regarding mean age, severity of underlying chronic respiratory disease, PaCO2, PaO2, and hemodynamic data on admission. P(a-em)CO2 and R were significantly different in PE+ and PE- patients at 12 +/- 6.9 torr compared to 1 +/- 2.4 torr and at 28 +/- 14.8% compared to 2 +/- 6.2% (p less than .001), respectively. The positive predictive value of the test was 74%, but the negative predictive value 100% and the specificity was 65%, but sensitivity was 100%.(ABSTRACT TRUNCATED AT 250 WORDS)


Mocravimod, a Selective Sphingosine-1-Phosphate Receptor Modulator, in Allogeneic Hematopoietic Stem Cell Transplantation for Malignancy.
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains the sole curative option for patients with acute myelogenous leukemia. Outcomes are limited by leukemia relapse, graft-versus-host disease (GVHD), and abnormal immune reconstitution. Mocravimod (KRP203) is an oral sphingosine-1-phosphate receptor (S1PR) modulator that blocks the signal required by T cells to egress from lymph nodes and other lymphoid organs. Mocravimod retains T cell effector function, a main differentiator to immunosuppressants. In preclinical models, mocravimod improves survival by maintaining graft-versus-leukemia (GVL) activity while reducing GVHD. In patients undergoing allo-HSCT for hematological malignancies, mocravimod is postulated to prevent GVHD by redistributing allogeneic donor T cells to lymphoid tissues while allowing a sufficient GVL effect in the lymphoid, where malignant cells usually reside. The primary objective of this study was to assess the safety and tolerability of mocravimod in patients undergoing allo-HSCT for hematologic malignancies. Secondary objectives were to determine the pharmacokinetic profiles of mocravimod and its active metabolite mocravimod-phosphate in this patient group, as well as to assess GVHD-free, relapse free survival at 6 months after the last treatment. In this 2-part, single- and 2-arm randomized, open-label trial, we evaluated the safety, tolerability, and pharmacokinetics of mocravimod in allo-HSCT recipients (ClinicalTrials.gov identifier NCT01830010). Patients received either 1 mg or 3 mg mocravimod per day on top of standard of care GVHD prophylaxis with either cyclosporine A/methotrexate or tacrolimus/methotrexate. We found that mocravimod can be safely added to standard treatment regimens in patients with hematologic malignancies requiring allo-HSCT. Mocravimod resulted in a significant reduction of circulating lymphocyte numbers and had no negative impact on engraftment and transplantation outcomes. Our results indicate that mocravimod is safe and support a larger study to investigate its efficacy in a homogeneous acute myelogenous leukemia patient population undergoing allo-HSCT.


A familial spontaneous epileptic feline strain: a novel model of idiopathic/genetic epilepsy.
A spontaneous epileptic model of cats has not been described previously. Recently, we identified familial epileptic cats and investigated their clinical features. These epileptic cats are healthy except for the presence of recurrent seizures that are typically a focal limbic seizure with secondary generalization. Furthermore, generalized seizures were induced by vestibular stimulation in some cats. This spontaneous epileptic cat strain may be a valuable model for idiopathic/genetic epilepsy.


Regression of left ventricular hypertrophy during 10 years after valve replacement for aortic stenosis is related to the preoperative risk profile.
Previous studies have suggested that regression of hypertrophy may be the underlying determinant of longevity and left ventricular function after valve replacement (AVR) for aortic stenosis (AS). The potential for hypertrophy regression could therefore be related to the preoperative risk profile. Ninety-one consecutive patients with AS had a "project" Doppler-echo and radionuclide ventriculography in addition to the standard investigation programme prior to AVR with a disc valve (19-29mm, n=82), a caged ball valve (26-29mm, n=8), or a stented porcine valve (26mm, n=1); 49 (group A) were selected for a serial follow-up study while 42 served as controls (group B). Forty-two group A patients took part in a 1.5-year examination while 47 (26 group A, 21 group B) patients were studied at 10 years. Groups A and B were comparable as regards all pre- and intra-operative data including left ventricular mass index (LVMi). A previously developed preoperative prognostic index (PI) separated the patients into groups with low (n=23), intermediary (n=19) and high risk (n=49) with 10-year survivals of 87%, 58% and 43% (P<0.01). LVMi dropped from 202+/-58g/m(2)preoperatively to 152+/-45g/m(2)(P<0.0001) at 1.5 years, and 139+/-40g/m(2)(P<0.0001) at 10 years (three and six patients, respectively, with paravalvular leak or mitral regurgitation excluded). PI correlated with preoperative (r=0.51, P<0.001), 1.5-year (r=0.46, P<0.01), and 10-year LVMi (r=0.41, P<0.01). Also preoperative left ventricular ejection fraction correlated with the three LVMi measurements. Patients with systemic hypertension had higher LVMi at 1.5 years (193+/-42, n=6 vs 144+/-42, n=33, P<0.05) and 10 years (175+/-39, n=12 vs 124+/-31g/m(2), n=29, P<0.001). Patients with low, intermediary or high PI, excluding those with hypertension, had 1.5-year LVMi of 110+/-35 (n=8), 134+/-43 (n=9) and 164+/-33g/m(2)(n=16; P<0.01), respectively, and 10-year LVMi of 116+/-25 (n=17), 126+/-27 (n=6), and 146+/-41g/m(2)(n=6; P<0.05), respectively. There was no relation between LVMi at 1.5 or 10 years and peak or mean Doppler gradient, prosthetic valve size, or valve size index. Left ventricular hypertrophy regression for patients who survived up to 10 years after AVR for AS is dependent on the preoperative risk profile indicating that irreversible myocardial disease is the underlying factor. Systemic hypertension is an important factor in its own right.


Lymphatic drainage of the pericardium to the mediastinal lymph nodes.
The lymphatic drainage of the pericardium may have a prognostic value in lung cancer and its anatomical pattern was the purpose of this study. The lymphatic vessels of the pericardium were visualized on 12 fresh adult human cadavers by injection of a green colorant by means of the classical equipment for pedal lymphography. Seventeen injections were performed on the right side and 22 injections on the left side, visualizing 20 and 29 lymphatic pathways respectively. Whatever the side injected, the lymphatic vessels mainly directed toward the tracheobronchial nodes (n = 35) and less frequently toward the prepericardial lymphatic vessels and nodes (n = 14). Such results demonstrate the aggravating role played by the lymphatic drainage in the prognosis of the lung cancers invading the pericardium.


Elevated fractalkine in patients with obstructive sleep apnea hypopnea syndrome.
Obstructive sleep apnea hypopnea syndrome (OSAHS) has been increasingly linked to cardiovascular disease. Inflammatory processes associated with OSAHS may contribute to atherosclerosis in these patients. Fractalkine is a unique chemokine which has both adhesive and chemoattractant functions. We tested the hypothesis that OSAHS patients have increased fractalkine. We studied 20 patients (18 males and 2 females) with newly diagnosed OSAHS, who were free of other diseases, had never been treated for OSAHS, and were taking no medications. We compared fractalkine measurements in these patients to measurements obtained in 15 control subjects (14 males and 1 female) who were matched for age and body mass index, and in whom occult OSAHS was excluded. Plasma fractalkine levels were significantly higher in patients with OSAHS than in controls (463.15 ± 110.78 versus 364.67 ± 64.81 pg/mL, F = 2.58, P = 0.004). Fractalkine were associated with AHI (r = 0.756, P < 0.0001), lowest oxygen saturation (r = -0.466, P = 0.005), and mean oxygen saturation (r = -0.344, P = 0.043). Plasma fractalkine levels were significantly decreased in patients with OSAHS after four nights nCPAP (463.15 ± 110.78 versus 416.75 ± 97.67 pg/mL, P = 0.001). OSAHS is associated with elevated levels of fractalkine, a marker of inflammation related to atherosclerosis. The severity of OSAHS is proportional to the fractalkine level.


The feather touch rasp, a powered instrument for hump reduction.
Standard rhinoplasty procedure involves bony profile alignment with an osteotome, followed by profile refinement with a manual rasp. The entire bony hump may sometimes be addressed with a rasp. Manual rasps and osteotomes, however, can be traumatic instruments, resulting in tissue oedema and bruising. The feather touch rasp is one of the powered instruments developed in recent years to improve precision and technical ease while preventing tissue trauma. The powered rasp has been frequently used at our department in the last eighteen months for hump reductions. Retrospective evaluation of 72 rhinoplasty procedures performed in 68 patients between January 2004 and June 2005. A bony hump reduction was necessary in 52 of the 68 patients. 60% of the patients were male. The mean age was 30 years. The open rhinoplasty technique was used in 65% of the patients. All humps were addressed with the feather touch rasp only. Patients were seen 10 days, 4 weeks and three months after surgery. In one patient the nasal dorsum remained too high. We found one asymmetric nasal dorsum. Another patient had a low nasofrontal angle, creating a false impression of a remaining hump. In two patients, bony irregularities appeared a few months after the rhinoplasty procedure. Overall, most patients were satisfied with the results of the hump reduction. The feather touch rasp makes safe and gradual bony hump reduction possible, with fragments being aspirated and the overlying skin being well protected.


(E)-1-(3,5-Di-meth-oxy-phen-yl)-3-(3-meth-oxy-phen-yl)prop-2-en-1-one.
In the title mol-ecule, C18H18O4, the C=C bond of the central enone group adopts a trans conformation. The relative conformation of the C=O and C=C bonds is s-cisoid. The dihedral angle between the planes of the benzene rings is 29.49 (12)°. In the crystal, weak C-H⋯O hydrogen bonds link the mol-ecules into chains along [010].


Comparison of donepezil and memantine for protective effect against amyloid-beta(1-42) toxicity in rat septal neurons.
Donepezil, a potent acetylcholinesterase (AChE) inhibitor and memantine, an N-methyl-d-aspartate (NMDA) receptor antagonist, have been used for the treatment of Alzheimer's disease (AD), and both of them have been shown to have neuroprotective action against glutamate excitotoxicity. However, it is not known whether donepezil and memantine similarly exert neuroprotective effects against amyloid-beta peptide(1-42) [Abeta(1-42)] toxicity in cholinergic neurons. Therefore, in the present study we compared the neuroprotective effects of donepezil and memantine against Abeta(1-42) toxicity in rat cultured septal cholinergic neurons, because deficit in cholinergic neurotransmission is a major feature in AD, and medial septal cholinergic neurons are known to degenerate in AD patients. Septal neuronal cells were cultured for 7 days and then 5 micromol/L of Abeta(1-42) was added to the medium for 48 h. Measurement of the efflux of lactate dehydrogenase (LDH) indicated that septal neuronal cells were highly susceptible to Abeta toxicity and relatively resistant to NMDA toxicity. Donepezil concentration-dependently reduced the LDH efflux induced by Abeta(1-42), and the effect was significant at 1 micromol/L and above. NMDA receptor antagonists, memantine and MK-801, did not show a significant neuroprotective effect against Abeta(1-42) toxicity. It is concluded that the neuroprotective effect of donepezil against Abeta(1-42) toxicity is not mediated by interference with the NMDA-mediated excitotoxic process, and that donepezil may be more effective than memantine against cholinergic neuronal damage induced by Abeta(1-42) exposure.


Antihepatotoxic activity of icariin, a major constituent of Epimedium koreanum.
In an attempt to identify compounds with antihepatotoxic activity, carbon tetrachloride-induced cytotoxicity in primary cultured rat hepatocytes has been adopted as a screening system. Using this screening system, an antihepatotoxic compound from the aerial parts of Epimedium koreanum has been isolated. This compound, icariin, is a flavonol glycoside. Its antihepatotoxic activity was first evaluated by measuring the release of glutamic pyruvic transaminase and sorbitol dehydrogenase from CCl4-intoxicated rat hepatocytes into the culture medium. Icariin significantly reduced the level of glutamic pyruvic transaminase and sorbitol dehydrogenase released resulting in a 76% protection from toxicity at concentration ranges from 1 microM to 20 microM. The antihepatotoxic activity of icariin was also estimated by the determination of total cytochrome P-450 content and glutathione-S-transferase activity in the CCl4-intoxicated hepatocytes.


On exposure and response relationships for health effects associated with exposure to vehicular traffic.
This work examines various metrics and models that have been used to estimate long-term health effects of exposure to vehicular traffic. Such health impacts may include effects of air pollution due to emissions of combustion products and from vehicle or roadway wear, of noise, stress, or from socioeconomic effects associated with preferred residential locations. Both categorical and continuous exposure metrics are considered, typically for distances between residences and roadways, or for traffic density or intensity. It appears that continuous measures of exposure tend to yield lower risk estimates that are also more precise than categorical measures based on arbitrary criteria. The selection of appropriate exposure increments to characterize relative risks is also important in comparing pollutants and other agents. Confounding and surrogate variables are also important issues, since studies of traffic proximity or density cannot identify the specific agents related to traffic exposures that might be responsible for the various health endpoints that have been implicated. Studies based on ambient air quality measurements are necessarily restricted to species for which data are available, some of which may be serving as markers for the actual agents of harm. Studies based on modeled air quality are limited by the accuracy of mobile source emission inventories, which may not include poorly maintained (high emitting) vehicles. Additional exposure modeling errors may result from precision limitations of geocoding methods. Studies of the health effects of traffic are progressing from establishing the existence of relationships to describing them in more detail, but effective remedies or control strategies have generally not yet been proposed in the context of these epidemiological studies. Resolution of these dose-response uncertainties is important for the development of effective public health strategies for the future.


Learning from the COVID-19 pandemic: health care disturbances and telemedicine as an alternative rheumatology practice in Indonesia.
Coronavirus disease 2019 (COVID-19) affects health care services. Our aim was to assess health care disruptions, treatment interruptions, and telemedicine reception regarding autoimmune rheumatic diseases (ARDs) in Indonesia. A cross-sectional population online-based questionnaire was conducted in Indonesia from September to December 2021. A total of 311 ARD patients were included, of whom 81 (26.0%) underwent consultations via telemedicine during the COVID-19 pandemic. The respondents showed increased concern about their susceptibility to COVID-19 (score of 3.9/5). Approximately 81 (26.0%) avoided hospital visits, and 76 (24.4%) stopped taking the medication without medical advice. Respondents' concerns correlated with their social distancing behaviors (p value 0.000, r 0.458). Respondent concerns, behaviors, and blocked access to the hospital during the pandemic were associated with avoiding hospital visits (p value 0.014; 0.001; 0.045; 0.008). Sex was associated with stopping medication (p value 0.005). In multivariate analysis, blocked access and sex remained significant. Approximately 81 (26%) respondents who used telemedicine services during the COVID-19 pandemic as an alternative medical consultation method showed high satisfaction (3.8/5). Health care disruptions and treatment interruptions were affected by patients' internal and external factors during the COVID-19 pandemic. Telemedicine may be the best option to address barriers to health care access in Indonesia's rheumatology practice during and after the pandemic situation.


The correlation between microscopical examination and erythrocyte band 3 (AE1) gene deletion in South-east Asian ovalocytosis.
South-east Asian ovalocytosis status was determined by microscopical examination of peripheral blood samples collected from 137 individuals in Papua New Guinea. The examination was performed separately by 2 microscopists, one of whom was very experienced in examining peripheral blood films for the diagnosis of south-east Asian ovalycytosis and the other was recently trained. The samples were also analysed by polymerase chain reaction (PCR) to determine ovalocytosis status by demonstrating a 27 base pair deletion in erythrocyte band 3 protein of the affected individuals. The microscopists were unaware of each other's results and of those obtained by PCR. Generally, there was very good agreement between the results obtained by both microscopists and the PCR. Although there was considerable inter-observer variation in the final ovalocyte count between the 2 microscopists, this did not affect their ability to discriminate between ovalocytic and normocytic individuals. Taking the PCR results as the standard, for the first, more experienced observer, the most efficient ovalocyte count cut-off point was around 50%. At this ovalocyte count the sensitivity and specificity of microscopical examination were 93.6% and 92.2%, and the positive and negative predictive values 86.3% and 96.5%, respectively. The second microscopist generally underscored the ovalocyte counts and his most efficient cut-off point was 20%, with sensitivity and specificity of 85.1% and 93.3% and positive and negative predictive values of 87.0% and 92.3%, respectively.


Assessment of bacterial endosymbiont diversity in Otiorhynchus spp. (Coleoptera: Curculionidae) larvae using a multitag 454 pyrosequencing approach.
Weevils of the genus Otiorhynchus are regarded as devastating pests in a wide variety of horticultural crops worldwide. So far, little is known on the presence of endosymbionts in Otiorhynchus spp.. Investigation of endosymbiosis in this genus may help to understand the evolution of different reproductive strategies in these weevils (parthenogenesis or sexual reproduction), host-symbiont interactions, and may provide a future basis for novel pest management strategy development. Here, we used a multitag 454 pyrosequencing approach to assess the bacterial endosymbiont diversity in larvae of four economically important Otiorhynchus species. High-throughput tag-encoded FLX amplicon pyrosequencing of a bacterial 16S rDNA fragment was used to characterise bacterial communities associated with different Otiorhynchus spp. larvae. By sequencing a total of ~48,000 PCR amplicons, we identified 49 different operational taxonomic units (OTUs) as bacterial endosymbionts in the four studied Otiorhynchus species. More than 90% of all sequence reads belonged either to the genus Rickettsia or showed homology to the phylogenetic group of "Candidatus Blochmannia" and to endosymbionts of the lice Pedicinus obtusus and P. badii. By using specific primers for the genera Rickettsia and "Candidatus Blochmannia", we identified a new phylogenetic clade of Rickettsia as well as "Candidatus Nardonella" endosymbionts in Otiorhynchus spp. which are closely related to "Candidatus Blochmannia" bacteria. Here, we used multitag 454 pyrosequencing for assessment of insect endosymbiotic communities in weevils. As 454 pyrosequencing generates only quite short sequences, results of such studies can be regarded as a first step towards identifying respective endosymbiotic species in insects. In the second step of our study, we analysed sequences of specific gene regions for a more detailed phylogeny of selected endosymbiont genera. As a result we identified the presence of Rickettsia and "Candidatus Nardonella" endosymbionts in Otiorhynchus spp.. This knowledge is an important step in exploring bacteria-insect associations for potential use in insect pest control.


Pro-inflammatory cytokine response of the fluid contents of spermatoceles and epididymal cysts.
The aim of the study was to determine the cellular contents and concentrations of interleukin 6 (IL-6), interleukin 8 (IL-8) and tumour necrosis factor alpha (TNF-alpha) in fluids of patients with spermatocele or epididymal cyst. Twenty-five symptomatic patients, 14 with epididymal cysts and 11 with spermatoceles, were included in the study. Fluids were obtained during surgical excision of the cysts and cytological smears were stained with May-Gruenwald-Giemsa to establish cell components. The concentrations of IL-6, IL-8 and TNF-alpha were measured by chemiluminescent immunometric assay. Cytological analysis of the fluids demonstrated various sperm forms ranging from immature germ cells to degenerated spermatozoa without inflammatory cells such as neutrophils and macrophages. The concentrations (mean+/- SEM, pg/mg protein) of IL-6, IL-8 and TNF-alpha were 13.52 +/- 1.40, 22.20 +/- 2.43, 3.51 +/- 1.43 in spermatocele fluids and 5.76 +/- 0.48, 11.57 +/- 1.89, 2.53 +/- 0.41 in epididymal cyst fluids. Both IL-6 and IL-8 concentrations in the spermatocele group were higher than in the epididymal cyst group (P < 0.0001). There were no differences in TNF-alpha concentrations between the groups (P > 0.05). These findings indicate that local production of pro-inflammatory cytokines is involved in cyst formation. The presence of immunologic activation in these fluids advocates a policy of selective surgical intervention in patients with spermatocele or epididymal cyst.


Increased prevalence of lymphoid tissue inducer cells in the cerebrospinal fluid of patients with early multiple sclerosis.
Inflammatory cytokines produced by cells of the immune system are believed to play a central role in the pathogenesis of multiple sclerosis (MS). Innate lymphoid cells (ILCs) have been shown to produce and secrete a wide range of the cytokines involved in MS pathogenesis; however, a possible implication of ILCs in MS development and disease progression has not been investigated. With this study, we aimed to clarify a potential role of ILCs in the early stages of MS. Using flow cytometry, we analysed the prevalence and phenotype of ILCs in the cerebrospinal fluid (CSF) of patients experiencing their first or second demyelinating event. We found a substantial increase in both frequency and number of ILCs, in particular the LTi subset, as compared to healthy controls. We also found an association between CSF pleocytosis and an increased frequency of LTi cells in the CSF, suggesting a favoured recruitment of blood derived LTi cells. Our data suggests a role for ILCs, and in particular the LTi subset, in the early stages of MS. This finding represents an important contribution to the understanding of early inflammation in MS, and adds new knowledge beneficial for future MS therapies.


Effects of myo-inositol vs. metformin on hormonal and metabolic parameters in women with PCOS: a meta-analysis.
Polycystic Ovary Syndrome is the most prevalent hormonal disorder in females. Over the years, metformin (MET) has become the first-line choice of treatment; however, due to its gastrointestinal side effects, a more recent drug, myo-inositol (MI), has been introduced. We aim to conduct a systematic review and meta-analysis to compare the effects of MET and MI on hormonal and metabolic parameters. Authors extensively searched PubMed, Scopus, Cochrane Library, Google Scholar, and Web of Science for randomized clinical trials (RCTs) until August 2021. Eight (n = 8) articles were included, with a total sample size of 1088, of which 460 patients received MET, 436 received MI, and 192 received a combination of both. Standard mean differences (SMDs) and Confidence Intervals (CIs) were used for data synthesis, and forest plots were made using Review Manager 5.4 for Statistical Analysis using the random-effect model. The meta-analysis indicates that there is no significant difference between MET and MI in terms of their effects on BMI (SMD = 0.16, 95% CI: - 0.11 to 0.43, p = 0.24), fasting insulin (SMD = 0.00, 95% CI: - 0.26 to 0.27, p = 0.97), fasting blood sugar (SMD = 0.11, 95% CI: - 0.31to 0.53, p = 0.60), HOMA index (SMD = 0.09, 95% CI: - 0.20 to 0.39, p = 0.50), and LH/FSH (SMD = 0.20, 95% CI: - 0.24 to 0.64, p = 0.37). BMI, fasting blood sugar, and LH/FSH ratio reported moderate heterogeneity because of the varying number of study participants. Our meta-analysis comparing hormonal and metabolic parameters between MET and MI did not show much significant difference, indicating both drugs are equally beneficial in improving metabolic and hormonal parameters in patients with PCOS.


Biodiversity can benefit from climate stabilization despite adverse side effects of land-based mitigation.
Limiting the magnitude of climate change via stringent greenhouse gas (GHG) mitigation is necessary to prevent further biodiversity loss. However, some strategies to mitigate GHG emission involve greater land-based mitigation efforts, which may cause biodiversity loss from land-use changes. Here we estimate how climate and land-based mitigation efforts interact with global biodiversity by using an integrated assessment model framework to project potential habitat for five major taxonomic groups. We find that stringent GHG mitigation can generally bring a net benefit to global biodiversity even if land-based mitigation is adopted. This trend is strengthened in the latter half of this century. In contrast, some regions projected to experience much growth in land-based mitigation efforts (i.e., Europe and Oceania) are expected to suffer biodiversity loss. Our results support the enactment of stringent GHG mitigation policies in terms of biodiversity. To conserve local biodiversity, however, these policies must be carefully designed in conjunction with land-use regulations and societal transformation in order to minimize the conversion of natural habitats.


A Retrospective Study on Endodontic Treatment Outcomes in Patients With Special Needs.
Outcomes of endodontic treatment in patients with special needs are not fully understood, leading to reluctance in performing endodontic treatment. This retrospective cohort study evaluated the periapical healing and tooth survival in patients with special needs who had permanent teeth endodontically treated in a hospital setting between 1998 and 2019. Data characterization was performed using univariate analyses. Logistic and Cox regression analyses (P < .05) were performed to identify the odds ratio (OR) and hazard ratio (HR) of the potential prognostic factors, respectively. These predictors were chosen from previous outcome studies in the general population and patients with special needs. A total of 61 patients (108 teeth) met the inclusion criteria. The most common treatment modality was initial therapy with 81.5% of cases being managed without a need for general anesthesia. Healing rate for initial endodontic treatment was 89.9% and was affected by no restoration at follow-up (OR = 0.15; 95% confidence interval [CI], 0.12-0.71; P = .02). After a mean follow-up of 79.36 ± 59.6 months, the survival rate was 73% and was correlated with gender (HR for males = 0.3; 95% CI, 0.1-0.6; P < .05) and age (HR for patients >45 years = 3.7; 95% CI, 1.7-8.2; P < .05). The most common reason for tooth extraction was unrestorable tooth fracture. Endodontic therapy without a need for general anesthesia is a viable treatment option for patients with special needs. If the overall patient condition permits, then treatment can be administered by community dentists rather than waiting for a patient to be seen in a hospital setting.


Risk prediction of sleep disturbance in clinical nurses: a nomogram and artificial neural network model.
Sleep disturbance occur among nurses at a high incidence. To develop a Nomogram and a Artificial Neural Network (ANN) model to predict sleep disturbance in clinical nurses. A total of 434 clinical nurses participated in the questionnaire, a cross-sectional study conducted from August 2021 to June 2022.They were randomly distributed in a 7:3 ratio between training and validation cohorts.Nomogram and ANN model were developed using predictors of sleep disturbance identified by univariate and multivariate analyses in the training cohort; The 1000 bootstrap resampling and receiver operating characteristic curve (ROC) were used to evaluate the predictive accuracy in the training and validation cohorts. Sleep disturbance was found in 180 of 304 nurses(59.2%) in the training cohort and 80 of 130 nurses (61.5%) in the validation cohort.Age, chronic diseases, anxiety, depression, burnout, and fatigue were identified as risk factors for sleep disturbance. The calibration curves of the two models are well-fitted. The sensitivity and specificity (95% CI) of the models were calculated, resulting in sensitivity of 83.9%(77.5-88.8%)and 88.8% (79.2-94.4%) and specificity of83.1% (75.0-89.0%) and 74.0% (59.4-84.9%) for the training and validation cohorts, respectively. The sleep disturbance risk prediction models constructed in this study have good consistency and prediction efficiency, and can effectively predict the occurrence of sleep disturbance in clinical nurses.


Reduction of the Fe(III)-tyrosyl radical center of Escherichia coli ribonucleotide reductase by dithiothreitol.
The active form of protein B2, the small subunit of ribonucleotide reductase from Escherichia coli, contains a binuclear ferric center and a free radical localized to tyrosine 122 of the polypeptide chain. MetB2 is an inactive form that lacks the tyrosine radical but retains the Fe(III) center. We earlier reported (Fontecave, M., Eliasson, R., and Reichard, P. (1989) J. Biol. Chem. 264, 9164-9170) that enzymes from E. coli interconvert B2 and metB2, possibly as part of a regulatory mechanism. Introduction of the tyrosyl radical into metB2 occurred in two steps: first, the Fe(III) center was reduced to Fe(II), generating "reduced B2"; next oxygen regenerated non-enzymatically both Fe(III) and the tyrosyl radical. Here we demonstrate that dithiothreitol (DTT) between pH 8 and 9.5 also slowly converts metB2 to B2 in the presence of oxygen. Also in this case the reaction occurs stepwise with reduced B2 as an intermediate. DTT reduces Fe(III) of both metB2 and B2. In the latter case this reaction is accompanied by the immediate loss of the tyrosyl radical. Our results indicate that the tyrosyl radical can exist only in the presence of an intact Fe(III) center. In reduced B2 iron is loosely bound to the protein, dissociates on standing and is readily removed by chelating agents. Binding decreases at higher pH. Loss of iron from reduced B2 explains why ferrous iron stimulates and iron chelators inhibit reactivation of metB2. We propose that the reactivation of mammalian ribonucleotide reductase by DTT (Thelander, M., Gräslund, A., and Thelander, L. (1983) Biochem. Biophys. Res. Commun. 110, 859-865) may proceed via a mechanism similar to the one found here for E. coli protein B2.


Patterns of internet use by parents of children attending a pediatric surgical service.
The internet has revolutionised the way we search for information. We determined the level of internet use by parents of children attending general surgical services and identified trends in online information-seeking behaviour. A questionnaire based on the work by Boston and Tassone was distributed to parents attending both the day surgical units and surgical outpatients department in a paediatric tertiary referral centre. There were 214 (82.3 %) questionnaires returned, with 82 (38.3 %) of respondents having searched the internet regarding their child's surgical issue. Access to a smartphone, a university education and private health insurance were factors that positively influenced online searching (p < 0.005). Of those respondents who searched the internet, 42 (51 %) felt that information they found online was understandable, while only 14 (17 %) admitted to online sourced information influencing the treatment decisions they had made for their children. When asked to rank information sources on Likert-type scales in terms of importance; parents ranked the surgeon as most important (mean = 4.73), whilst the internet ranked lowest (mean = 3.02). We demonstrated significant use of the internet amongst those attending paediatric general surgical services. Clinician sourced information remains important, however we should engage with patients to utilise this vast resource effectively.


Are black circles attentive fields? A reply to Bross, Blair, and Longtin.
It is argued that the experiment carried out by Bross et al is not a valid test of assimilation theory because they confuse a graphic representation of a constant with the construct itself and assume that changing the former will alter the latter.


Nucleic acid distribution pattern in avian erythrocytes and mammalian lymphocytes: comparative studies by fluorescence microscopy and digital imaging analytical techniques.
Nucleated erythrocytes of healthy domestic chicken and ducks, and lymphocytes of healthy Sprague Dawley rats were evaluated for nucleic acid distribution pattern, employing light and fluorescence microscopy procedures, as well as digital imaging analytical methods. The results demonstrate a unique organization of nuclear DNA of mature chicken and duck erythrocytes, as well as immature duck erythrocytes, as delineated spherical nuclear bodies that mostly corresponded with euchromatin zones of the cells in routine Wright-stain blood smears. The nuclear DNA of the rat lymphocytes, on the other hand, was observed as a more diffuse green fluorescing nuclear areas, with punctate variably-sized diffuse areas of RNA red fluorescence. RNA red color fluorescence was also evident in the narrow cytoplasm of the lymphocytes, especially in large lymphocytes, in comparison with the cytoplasm of the mature avian erythrocytes that completely lacked any nucleic acid fluorescence. Nuclear RNA fluorescence was lacking in the mature chicken erythrocytes, compared with those of the mature and immature duck erythrocytes as well as lymphocytes of both avian and rats blood. The significance of these findings lies in the establishment of normal benchmarks for the nuclear and cytoplasmic nucleic acid pattern in eukaryotic cells. These normal benchmarks become valuable in rapid diagnostic situations associated with pathologies, such as the presence of viral nuclear and cytoplasmic inclusion bodies that can alter the nucleic acid pattern of the host cells, and in conditions of cellular abnormal protein aggregations. Variability of cellular nucleic acid pattern can also aid in prognostic assessments of neoplastic conditions.


Single and multispecies microalgae toxicological tests assessing the impact of several BPA analogues used by industry.
BPA is a hazard for human and environmental health and recently BPA was added to the Candidate List of substances of very high concern by European Chemical Agency (ECHA). In accordance with this proposal, the authorities have encouraged the replacement of BPA by BPA analogues; however, little is known about the impact of these compounds on the environment. Due to this situation five BPA analogues (BPS, BPAP, BPAF, BPFL and BPC) were chosen in order to study their effects on marine primary producers. Three marine microalgae species (Phaeodactylum tricornutum, Tetraselmis suecica and Nannochloropsis gaditana) were selected for single and multispecies tests concerning the ecotoxicological effects of these BPA analogues. Microalgae were exposed to BPs over 72 h at different dosages (5, 20, 40, 80, 150 and 300 μM). Responses such as: growth, ROS production, cell complexity, cell size, autofluorescence of chlorophyll a, effective quantum yield of PSII and pigment concentrations were assessed at 24, 48 and 72 h. The results revealed that BPS and BPA showed lower toxicity to microalgae in comparison with BPFL > BPAF > BPAP and >BPC for the endpoints studied. N. gaditana was the least sensitive microalgae in comparison to P. tricornutum and T. suecica. However, a different trend was found in the multispecies tests where T. suecica dominated the microalgae community in relation to N. gaditana and P. tricornutum. The results of this work revealed for first time that present day BPA analogues are a threat and not a safe substitute for BPA in terms of the marine phytoplanktonic community. Therefore, the results of their impact on aquatic organisms should be shared.


Effect of hypocholesterolemia on cholesterol synthesis in small intestine of diabetic rats.
Studies by our and other laboratories have demonstrated that cholesterol synthesis is increased in the small intestine of insulinopenic diabetic animals. In normal animals, many factors have been shown to regulate cholesterol synthesis in the small intestine, including changes in plasma cholesterol levels. The purpose of this study was to determine the effect of lowering plasma cholesterol levels on small intestine cholesterol synthesis in streptozocin-induced diabetic rats. In diabetic rats, 4-aminopyrazolo[3,4-d]pyrimidine (4-APP)-induced hypocholesterolemia (plasma cholesterol levels less than 20 mg/dl) resulted in a 2.5-fold increase in small intestine cholesterol synthesis, which was most marked in the distal small intestine, decreasing proximally. In the distal small intestine the incorporation of 3H2O into cholesterol was 0.28 +/- 0.04 mumol.h-1.g-1 in diabetic rats versus 1.60 +/- 0.38 in diabetic rats administered 4-APP (P less than .01). This stimulation of cholesterol synthesis occurred in the upper villus, middle villus, and crypt cells isolated from the middle intestine of the 4-APP-treated diabetic animals. In agreement with these observations, "functional hypocholesterolemia" due to Triton WR-1339 administration also stimulated cholesterol synthesis 2.5-fold in the small intestine of normal and diabetic animals. In the distal small intestine, cholesterol synthesis was 0.43 +/- 0.10 mumol.h-1.g-1 in the diabetic rats versus 1.08 +/- 0.21 in diabetic rats treated with Triton WR-1339 (P less than .05). In both the 4-APP and Triton WR-1339 experiments, the response of the diabetic rats was similar to that observed in normal rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Non-carcinogenicity of capsaicinoids in B6C3F1 mice.
The carcinogenicity of a mixture of capsaicinoids (64.5% capsaicin and 32.6% dihydrocapsaicin) was examined in B6C3F1 mice. In a 13-week toxicity study, renal toxicity was observed in 1% capsaicinoid-treated males. Next, groups of 50 mice of each sex were given 0, 0.025, 0.083 or 0.25% capsaicinoids in powdered diet for 79 weeks and killed in week 83. Food intake was reduced in mice of all capsaicinoid-treated groups, especially females, because of the pungency of capsaicinoids, and inhibition of body weight gain was apparent in females. The numbers of tumour-bearing females in the high-dose groups were significantly lower than that in the controls, and the incidences of hepatocellular neoplasms in both sexes were negatively correlated with the dose of capsaicinoids (Cochran-Armitage trend test). Renal cell adenomas developed in one mouse each of 0.025 and 0.25% capsaicinoid-treated males. The incidences of other tumours were similar in the treated and control groups. Thus, the present study indicated that a mixture of capsaicinoids is not carcinogenic in B6C3F1 mice.


Matrine suppresses airway inflammation by downregulating SOCS3 expression via inhibition of NF-κB signaling in airway epithelial cells and asthmatic mice.
Matrine has been demonstrated to attenuate allergic airway inflammation. Elevated suppressor of cytokine signaling 3 (SOCS3) was correlated with the severity of asthma. The aim of this study was to investigate the effect of matrine on SOCS3 expression in airway inflammation. In this study, we found that matrine significantly inhibited OVA-induced AHR, inflammatory cell infiltration, goblet cell differentiation, and mucous production in a dose-dependent manner in mice. Matrine also abrogated the level of interleukin (IL)-4 and IL-13, but enhanced interferon (IFN)-γ expression, both in BALF and in lung homogenates. Furthermore, matrine impeded TNF-α-induced the expression of IL-6 and adhesion molecules in airway epithelial cells (BEAS-2B and MLE-12). Additionally, we found that matrine inhibited SOCS3 expression, both in asthmatic mice and TNF-α-stimulated epithelial cells via suppression of the NF-κB signaling pathway by using pcDNA3.1-SOCS3 plasmid, SOCS3 siRNA, or nuclear factor kappa-B (NF-κB) inhibitor PDTC. Matrine suppresses airway inflammation by downregulating SOCS3 expression via inhibition of NF-κB signaling in airway epithelial cells and asthmatic mice.


The impact of functional LIG4 polymorphism on platinum-based chemotherapy response and survival in non-small cell lung cancer.
DNA repair capacity is correlated with the sensitivity of cancer cells toward platinum-based chemotherapy. The aim of this study was to investigate whether single-nucleotide polymorphisms (SNPs) in DNA repair genes NBS1, LIG4, and RAD51 were correlated with tumor response in advanced non-small cell lung cancer (NSCLC) patients in a Chinese population who received platinum-based chemotherapy. The treatment outcomes of 146 advanced NSCLC patients who were treated with platinum-based chemotherapy were evaluated. The polymorphic status of three SNPs was determined by genotyping via the polymerase chain reaction-restriction fragment length polymorphism method. Forty-five patients in the group with the CC genotype (45/90) showed a good response to treatment, while only 18 patients in the CT+TT group (18/55) showed a good response, indicating a substantial differences in the chemotherapy response rate based on the LIG4 Thr9Ile polymorphism (P = 0.042). Patients with the GG genotype for the NSB1 Glu185Gln polymorphism were more sensitive to platinum-based chemotherapy compared with patients with either the CG or CC genotype (P = 0.001). Kaplan-Meier analysis of all patients showed a significant association between the LIG4 Thr9Ile CC polymorphism and superior progression-free survival and overall survival (log-rank P = 0.045 and 0.031, respectively). However, there were no significant differences in survival based on the LIG4 Thr9Ile or the RAD51 135G>C polymorphisms. Polymorphisms in the NSB1 and LIG4 genes may be a predictive marker for treatment response and for advanced NSCLC patients in stage IIIB + IV. The CC genotype of the LIG4 Thr9Ile polymorphism may also serve as an independent prognosis factor.


Crystallization and preliminary X-ray investigation of barstar, the intracellular inhibitor of barnase.
Crystals of barstar, the intracellular inhibitor of the extracellular ribonuclease produced by Bacillus amyloliquefaciens (barnase), were obtained through vapor phase equilibration using the hanging drop technique. Three crystal forms have been characterized. Forms I and II, crystallized either in potassium phosphate or sodium citrate, are tetragonal; they exhibit a superstructure along the c-axis. Form III crystals, suitable for a high resolution structure determination, were grown from 55-65% ammonium sulfate. This crystal form is hexagonal and diffracts to at least 2 A resolution at a synchrotron radiation source. It belongs to the hexagonal space group P6, with unit cell dimensions a = b = 143.6 A, c = 35.6 A. There are four molecules of barstar in the asymmetric unit. X-ray data have been collected to 2.2 A Bragg spacing. The structure determination is underway in order to analyze conformational changes of barstar upon complexation with barnase.


Efficacy of PD-1 inhibitors for colorectal cancer and polyps in Lynch syndrome patients.
Programmed death-1 (PD-1) inhibitor is effective for colorectal cancer (CRC) with deficient mismatch repair (dMMR) or high microsatellite instability (MSI-H). We aimed to explore its effects on CRCs and colonic polyps in Lynch syndrome (LS) patients. LS patients with CRC who had evaluable tumours and received at least 2 cycles of PD-1 inhibitors were retrospectively included. PD-1 inhibitors were given as a monotherapy or in combination with other therapies, including anticytotoxic T-lymphocyte-associated antigen-4 treatment, radiotherapy, chemotherapy, and targeted therapy. Correlations of treatment responses with clinicopathological characteristics and genomic profiles were analysed. A total of 75 LS patients were included, with a median age of 39 years. The median duration of follow-up was 27 months (range, 3-71). The objective response rate (ORR) was 70.7%, including 28.0% (n = 21) complete responses and 42.7% (n = 32) partial responses. Four of five cases of LS CRCs displaying proficient MMR (pMMR) or microsatellite stable (MSS) were not responsive. Mucinous/signet-ring cell differentiation was associated with a lower ORR (P = 0.013). The 3-year overall survival and progression-free survival were 91.2% and 82.2%, respectively. A polyp was detected in 26 patients during surveillance. Seven adenomas disappeared after treatment, and they were all larger than 7 mm. PD-1 inhibitors are highly effective for dMMR and MSI-H LS CRCs, but not for pMMR or MSS LS CRCs or mucinous/signet-ring cell CRC. Large LS adenomas may also be eliminated by anti-PD-1 treatment. Due to the privacy of patients, the related data cannot be available for public access but can be obtained from Pei-Rong Ding (email@example.com) upon reasonable request. The key raw data have been uploaded to the Research Data Deposit public platform (www.researchdata.org.cn).


Quantitative characterization of pore structure of cellulose gels with or without bound protein ligand.
Structural properties of cellulose gels Perloza MT, materials designed for the preparation of chromatographic adsorbents and immobilized biocatalysts, having a different content of polymer were investigated using a batch solute exclusion method. A homologous set of dextrans with a wide range of molecular weights was used to probe the pore accessibility of the gel particles. It was found that all gels possessed a bimodal pore structure where macropores were fully accessible to all dextrans whereas the solute partitioning depending on the molecule size occurred in the micropores of the swollen polymer network. The macropore and micropore fractions of the gels were estimated from the masses of total water and water accessible to the largest solute. The macropore fraction decreased with the gel polymer content. It was 0.57 at the gel containing 8% of polymer but only 0.22 at the gel with 38% of polymer. The micropore fraction varied from 0.38 to 0.47. The mass of accessible water for each solute was used to calculate the particle and gel-phase partition coefficients. The dependence of the latter quantity on the solute hydrodynamic radius was successfully fitted with the Ogston model. Bovine serum albumin that was used as a model protein ligand blocked almost all gel-phase pores of the gel with the highest polymer content whereas it little affected the accessibility of other materials.


Proteoglycan synthesis in flat cell-free cultures of chick embryo retinal neurons and photoreceptors.
Extracellular matrix and cell surface proteoglycans are thought to play important roles in neural development and regeneration. Central nervous system proteoglycans have been isolated and characterized from rat and sheep brain and from chick neural retina. An experimental advantage offered by the latter tissue is that it is avascular and can be isolated free of connective tissue and pigment epithelium. Therefore, proteoglycans synthesized by this tissue are derived exclusively from neural cells. However, it has not yet been determined whether neurons and photoreceptors contribute to proteoglycan synthesis or whether these molecules are largely glial in origin. In the present study we have addressed this question using cultures of chick neural retinal cells free of flat, glial-like cells. Proteoglycans synthesized by cultures of retinal neurons, photoreceptors, and undifferentiated, process-free round cells from 8-day embryonic chick neural retina were metabolically labeled in vitro using [35S]sulfate and [3H]glucosamine as precursors. Radiolabeled proteoglycans accumulated in the medium, and could also be extracted from the cell layer by sequential treatments with Triton X-100 and with guanidine HCl. The proteoglycans were isolated by ion-exchange chromatography, and characterized by gel filtration chromatography and by susceptibility to degradation by enzymatic and chemical treatments. Overall, heparan sulfate proteoglycans were the predominant type of proteoglycan synthesized in vitro by the cultured neural retinal cells at this developmental stage. The medium and the Triton extract contained different proportions of both chondroitin sulfate and heparan sulfate proteoglycans, while heparan sulfate was the only proteoglycan recovered from the guanidine extract. These studies demonstrate that heparan sulfate and chondroitin sulfate proteoglycans are actively synthesized by cultures of neural retinal cells free of flat, glial-like cells.


[Caregiver in a mobile geriatric team, a new profession].
The caregiver plays a key role in the mobile geriatric team. She has a very varied activity. She performs geriatric assessments, evaluates toilets, transmits geriatric culture, maintains the city-hospital link, intervenes in residential facilities for dependent elderly people (Ehpad), performs post-emergency telephone interviews and provides training to paramedics. Testimonial.


Associations of moderate alcohol consumption with clinical and MRI measures in multiple sclerosis.
To examine the associations of alcohol consumption patterns with disability and brain injury in multiple sclerosis (MS) patients. This study included 423 subjects (272 MS patients, 151 healthy controls) participating in a study of clinical, environmental and genetic risk factors in MS. Disability was assessed with the Expanded Disability Status Scale (EDSS) and the MS Severity Scale (MSSS). Brain injury was assessed using the quantitative MRI measures of T2-lesion volume (T2-LV), T1-LV, normalized volumes of brain parenchyma (NBV), gray matter (NGMV) and lateral ventricle (NLVV). Information related to alcohol-consumption patterns was obtained with standardized questionnaire during an in-person interview. The associations of alcohol consumption variables with disability and MRI measures were assessed in regression analyses. The frequency of MS patients who did not consume alcohol after MS (19.4%) was higher than the frequency before MS (p<0.001). The EDSS, NGMV and NLVV exhibited a non-linear dependence on duration of alcohol consumption after MS onset: non-linear regression analyses indicated that EDSS and NLVV were lower and the NGMV was greater in MS patients who had consumed for a period of 15years or less after MS onset compared those who did not consume alcohol or consumed it for more than 15years. The duration of alcohol consumption is associated with disability and MRI measures in MS. Prospective, longitudinal studies of the role of alcohol in MS disease progression are warranted.


Eating attitudes and behaviors of pre- and postpubertal females: clues to the etiology of eating disorders.
Eating disorders are most likely to occur in females in early adolescence. In the hope of identifying the nature of the changes occurring across puberty which may predispose adolescents to eating disorders three groups, 30 prepubertal girls, 28 postpubertal girls, and 28 adult women matched on height and weight to the postpubertal group were compared for meal pattern characteristics and food intake regulation, as well as changes in eating attitudes and behaviors. They completed a set of psychometrics measuring their eating attitudes, body image, and disposition and also maintained 7-day diaries of everything they ingested, time of ingestion, and subjective and social conditions. Neither the overall intakes nor the meal sizes significantly differed between age groups. The prepubertal females ate the most relative to body weight, ate with more other people present and earlier in the day. They were relatively happy, calm, slightly hungry, unrestrained, satisfied with their bodies and depression was associated with lower intake. The postpubertal group, on the other hand, were relatively restrained, ate faster, and ate only 76.5% of the recommended amount. They had a more negative body image, greater dissatisfaction with their bodies, and greater restraint of eating than the prepubertal group, and depression was associated with higher intake. These changes may leave the young woman extremely vulnerable and under the appropriate conditions or with the appropriate personality type may lead to the development of an eating disorder.


Collaboration between child psychiatry and paediatrics: the state of the relationship in Norway.
In Norway, the first paediatric department was founded in Oslo 100 years ago. The first child psychiatric department was opened in 1950. To assess qualitative and quantitative aspects of child psychiatric liaison work, questionnaires were sent to the heads of 25 paediatric departments and 53 child psychiatric units. Scarce child psychiatric resources were spent in paediatrics. The average score for satisfaction with collaboration was moderate (5.5 on a 10-cm visual analogue scale) and agreement between the parties was modest. Improvement will involve development of a common language and a shared model for understanding the psychosocial aspects of acute and chronic childhood illnesses. In-service training for paediatricians in child psychiatry and vice versa may help. Both parties indicated a need for more training in consultation/liaison and in multi-professional assessments and therapeutic interventions with children with physical illnesses.


[Pathohistology of polyps of the gastrointestinal tract].
Polyps of the gastrointestinal tract encompass a variety of epithelial and non-epithelial tumour-like conditions. The most common polyps are epithelial lesions. In the upper gastrointestinal tract, reactive inflammatory changes and hyperplastic polyps dominate, whereas true neoplastic polyps, like adenomas, are much more common in the colorectum. In addition to neoplasias such as adenomas, non-neoplastic polyps such as hyperplastic polyps of the stomach may also be associated with an increased risk of malignancy. The risk of malignancy is determined by the histological subtype of polyp, as well as the size, presence and degree of dysplasia. The term "dysplasia" has been reintroduced for adenomas in the current 2019 World Health Organization (WHO) classification and replaces "intraepithelial neoplasia". A further change is the term "sessile serrated lesion" with and without dysplasia, which was formerly known as sessile serrated adenoma.


The contagion mechanism for the origin of sex.
The origin of sex is one of the least understood aspects of the evolution of sex. Recently it has been proposed that sex arose as a result of a contagious genetic element which acted so as to foster its own transmission. Two continuous-time deterministic models of this mechanism are analysed. The first model lacks population regulation. With this assumption, sex is always established in the population, predominating if the sexual form is viable. The second model includes population regulation, giving rise to evolutionary dynamics in which sex need not be established in the population. In addition, with the second model, when the sexual form is viable, it need not fix. Thus, the contagion mechanism can give rise to sex, but need not do so.


Influence of galantamine on vasomotor reactivity in Alzheimer's disease and vascular dementia due to cerebral microangiopathy.
Recent reports suggest that vascular factors play a crucial role in the development and progression of Alzheimer's disease. We aimed to assess vasomotor reactivity in patients with Alzheimer's disease and vascular dementia due to microangiopathy using transcranial Doppler sonography and near-infrared spectroscopy during a CO(2) exposition task. The normalized CO(2) reactivity assessed at the middle cerebral artery and the oxygenated and deoxygenated hemoglobin of the frontal cortex were obtained. To investigate the impact of cholinergic deficiency known for Alzheimer's disease on vasomotor reactivity, both groups were reinvestigated during treatment with the acetylcholine esterase inhibitor galantamine. Transcranial Doppler analysis revealed significantly reduced normalized CO(2) reactivity for Alzheimer's disease and vascular dementia. Vasomotor reactivity assessed by near-infrared spectroscopy was decreased in patients with vascular dementia, but not in Alzheimer's disease. Galantamine treatment showed a beneficial effect, normalizing these parameters close to age-matched control levels. Our results suggest that Alzheimer's disease is associated with a lack of vasomotor reactivity, which might be associated with disturbed autoregulation indicating a potential risk for a decreased protection of brain tissue against blood pressure changes. Additionally, a diminished increase of cortical oxygenated hemoglobin during the CO(2) test was apparent in patients with vascular dementia. Galantamine treatment influenced vascular reactivity in the CO(2) test, thus providing evidence for the cholinergic deficiency, thereby adding to vascular dysregulation in Alzheimer's disease, but also indicating an important role of cholinergic system dysfunction for vascular dementia.


The effect of thread length and location on extraction strengths of iliosacral lag screws.
Although iliosacral lag screws are an established technique for fixation of sacroiliac joint dislocation and sacral fractures, there is a paucity of data on the relative strength of fixation of screws in the sacral ala and body. The purpose of this study was to quantify and compare the extraction strength of cancellous screws in the sacral ala and body. Twelve fresh frozen cadaveric human pelves (mean age 76) were used to test the extraction strength of three groups of 7.0 mm cannulated cancellous screws: long-threaded in the sacral body, short-threaded in the body and short-threaded in the ala. The mean extraction strengths were 925, 374 and 71 newtons (or 92, 37, and 7 kg) respectively. The differences between the three groups were highly significant (all P < 0.0025). These data strongly recommend that the goal in iliosacral lag screw fixation should be to insert a long-threaded screw into the sacral body, if safely feasible. Fixation in the ala is inferior and should be avoided in the elderly.


Genetic control of collagen-induced arthritis in rats: the immune response to type II collagen among susceptible and resistant strains and evidence for multiple gene control.
We evaluated the cellular and humoral immune response to type II collagen and the relative susceptibility to type II collagen-induced arthritis (CIA) in 14 inbred rat strains representing six RT1 specificities and including four congenic strain pairs differing only at RT1, by using a standard protocol with native calf type II collagen as the immunogen: WF(RT1u) was a high responder and susceptible; RT1c , RT1a , and RT1l strains were intermediate responders and, on a strain basis, were variably susceptible or resistant; RT1b and RT1n strains were low responders and resistant to CIA. An intermediate to high immune response to type II collagen, although associated with CIA, was not sufficient for the induction of clinical arthritis. A high degree of cross-reactivity between calf and rat type II collagen was found. No selective, antigen-specific decrease in immune response to rat type II collagen as compared to calf type II collagen was found in the resistant strains. In six strains, resistance correlated with the expression of public Ia antigens cross-reactive with the public Ia antigens of BN. The non-MHC-linked BN genome also exerted a suppressive effect on the incidence and severity of CIA in strains carrying collagen-responder and CIA-susceptible RT1 alleles. Together, the data show that CIA in rats is under the control of multiple genes, both RT1-linked and nonlinked .


Leading a highly visible hospital through a serious reportable event.
Most preventable adverse events result from systemic causes, not human error. The senior vice president for patient care/chief nurse at a leading hospital recounts the unnecessary death of a patient and the investigation that followed. Citing the critical importance of a "just culture," this case study offers a blueprint for managing a serious reportable event.


Avulsion injury of the rat brachial plexus triggers hyperalgesia and allodynia in the hindpaws: a new model for the study of neuropathic pain.
In the present study, we sought to characterise a behavioural model of persistent peripheral neuropathic pain produced by avulsion of the right brachial plexus in rats. In addition, we compared the effects of avulsion with those of ligation or crush injury of the brachial plexus. Avulsion and, to a lesser extent, ligation and crushing of brachial plexus caused a long-lasting (up to 90 days) and highly reproducible mechanical hyperalgesia, in both ipsilateral and contralateral hindpaws. However, the same injury did not produce thermal hyperalgesia. The avulsion and, to a lesser extent, ligation and crushing of the brachial plexus elicited a significant and long-lasting (up to 90 days) ipsilateral and contralateral cold and mechanical allodynia. Furthermore, the brachial plexus injury caused a significant decrease in functional activity of the forepaws as assessed in the grasping strength test, but did not alter the locomotor activity of the rats in the open field test in comparison with control or sham groups. Taken together these results show that avulsion of the brachial plexus in rat produces persistent mechanical and cold allodynia and mechanical hyperalgesia, and might represent a valuable method for understanding the mechanisms underlying the aetiology of neuropathic pain.


A histochemical study of the innervation of the cerebral blood vessels in the domestic fowl.
Adrenergic and cholinergic nerves innervating the cerebral arteries of the domestic fowl were examined by specific histochemical techniques. The adrenergic nerve plexuses of the cerebral carotid system are markedly denser than those of other vertebrates observed by similar techniques. They form longitudinally elongated meshworks of fine fibres in the vascular wall of the arterial branches. Those innervating the vertebro-basilar system are less dense and more elongated, and, as the size of the artery diminishes, the fibres of the plexus become coarser. In the small pial and parenchymal arteries they are reduced to a few fibres running parallel to, or spiralling around the vascular axis. The cholinergic nerve plexuses are not as dense as the adrenergic system. The acetylcholinesterase activity is very weak, except in the plexuses innervating the cerebral carotid artery and proximal portion of the anterior and posterior rami. In the vertebro-basilar system, a few thick nerve bundles run alongside the blood vessels of the vertebral and basilar arteries. Cholinergic nerves enter the cranial cavity along the internal carotid, the vertebral and possibly the cerebroethmoidal arteries. Intracerebral capillaries and some arterioles are not innervated with cholinergic and adrenergic fibres of peripheral origin, but with ones arising from parenchymal nerve cells.


Kinetic effects of ATP, divalent metal ions and pH on chicken liver mevalonate 5-diphosphate decarboxylase.
The activity of chicken liver mevalonate 5-diphosphate decarboxylase was measured over a wide range of Mg2+ and ATP concentrations. It was found that free ATP activated the enzyme, whereas free Mg2+ had no effect on the enzyme activity. Computed analyses of free species concentrations and pH studies indicated that MgATP2- is the true substrate. The relative efficiencies of Mg2+, Mn2+, Cd2+, and Zn2+ as activating metal ions were evaluated in terms of V/Km for the corresponding (metal-ATP)2- complexes, and the relative ratios were: Mn2+ 100, Cd2+ 37, Mg2+ 14, Zn2+ 1.7. Inhibitory effects were demonstrated for all free divalent cations tested, except for Mg2+, and were in the order Zn2+ greater than Cd2+ greater than Mn2+.


Antagonists of CD117 (cKit) signaling inhibit mast cell accumulation in healing skin wounds.
Mast cells (MCs) have important functional roles in leukocyte recruitment, pain, and wound healing, and increased tissue resident MC function has been associated with several fibrotic diseases. Consequently, the study of MCs in situ can be a direct approach to studying the pharmacodynamic impact of MC-directed therapeutics in tissues. Here we describe an automated laser scanning cytometry assay that was used to characterize the kinetics of MC accumulation in healing skin wounds and to study the effect of inhibiting CD117 (cKit) signaling. The number of tryptase-positive MCs approximately doubled 14 days after cutaneous injury in nonhuman primates. Treatment of animals with anti-CD117 or imatinib mesylate (Gleevec) reduced MC accumulation at the edge of healing wounds in mice and nonhuman primates, respectively. In translating this MC assay to become a biomarker for human studies, no differences in dermal MC numbers were evident between genders, ages or body mass index from 20 healthy donors. These data suggest that skin is a practical and useful tissue for tracking pharmacodynamic effects of MC-directed therapies.


The cumulus cell gene expression profile of oocytes with different nuclear maturity and potential for blastocyst formation.
Gene expression in human ART cumulus cell (CC) has been related to oocyte maturity and competence but requires further validation. Expression dynamics were investigated in CC of oocytes at different maturational stages and with different developmental competence in a standard in vivo mouse superovulation model. Quantitative PCR analysis of Has2, Vcan, Sdc4, Alcam, Grem1, Ptgs1 and Ptgs2 in CC collected at regular time intervals from 0 to 24 h post hCG injection. Three expression patterns were observed each with strong regulation (4-230× differences). Immediately prior to ovulation CC of GVBD oocytes have 5× less Sdc4 and Ptgs1 and 5× more Ptgs2 when compared to the CC of freshly ovulated PB oocytes. When compared to the latter, the post-ovulatory aged PB oocytes had a 2× reduced blastocyst forming capacity and their CC expressed 2× more Sdc4 and 6× less Alcam. Morphologically identical cumulus oocyte complexes with different developmental competence can be differentiated by CC gene expression.


Estimating imprecision profiles in biochemical analysis.
We describe a computer program IMPROFIL which determines an imprecision profile of an analytical method from replicated measurements of samples. It calculates the variance function, the coefficient of variation, the power of definition, the critical limit, the limit of detection and the lower limit of quantification. The primary property, the variance function, is determined by two alternative methods: the conventional maximum approximate conditional likelihood method and the newly developed weighted absolute deviation method. For all quantities, confidence intervals are obtained using the bootstrap procedure. The program combines the use of robust numerical techniques, user-friendliness and integration into a spreadsheet program for data pre- and post-processing. The algorithms used are described in detail. Tests with synthetic data sets are used to validate the method and to establish its powers and limitations. Finally, its application to a practical analytical task (tumor marker CA 15-3 in human sera) is reported. For the method to yield a reliable estimate of the variance function and the derived properties, certain minimum requirements on the raw data must be met: They have to be spread throughout the concentration range of interest, there should not be less than three replicates per specimen, and there must be at least of the order of 25 (better at 50) specimens.


Caesarean section and the association with migraine: a retrospective register-linked HUNT population cohort study.
To evaluate the association between caesarean section and migraine in a population-based register-linked cohort study. Data from the population-based Nord-Trøndelag Health Studies (HUNT2 and HUNT3) were linked to information from the Norwegian Medical Birth Registry. 65 343 participants responded to the headache questions in any of the two HUNT studies. Only those answering the headache questions in HUNT2 or 3 and had information about mode of delivery in the Norwegian Medical Birth Registry (born after 1967) were included. Our final sample consisted of 6592 women and 4602 men, aged 19-41 years. ORs for migraine given caesarean section. Analyses were performed in multivariate logistic regression models. After adjusting for sex, age and fetal growth restriction, delivery by caesarean section was not associated with migraine later in life (OR 0.86, 95% CI 0.64 to 1.15). Delivery by caesarean section was associated with a reduced OR of non-migrainous headache (OR 0.77, 95% CI 0.60 to 0.99). No association was found between caesarean section and migraine in this population-based register-linked study.


Impact of autoclavation on baked-on siliconized containers for biologics.
Many pharmaceutical manufacturing units utilize pre-sterilized ready-to fill primary containers for parenterals. The containers may have been sterilized by the supplier via autoclavation. This process can change the physicochemical properties of the material and the subsequent product stability. We studied the impact of autoclavation on baked on siliconized glass containers for biopharmaceuticals. We characterized the container layers of different thickness before and after autoclavation for 15 min at 121 °C and 130 °C. Furthermore, we analyzed the adsorption of a mAb to the silicone layer and subjected filled containers to 12 weeks storage at 40 °C monitoring functionality and subvisible particle formation of the product. Autoclavation turned the initially homogenous silicone coating into an incoherent surface with uneven microstructure, changed surface roughness and energy, and increased protein adsorption. The effect was more pronounced at higher sterilization temperatures. We did not observe an effect of autoclavation on stability. Our results did not indicate any concerns for autoclavation at 121 °C for safety and stability of drug/device combination products using baked-on siliconized glass containers.


IL-1beta treatment does not co-ordinately up-regulate mPGES-1 and COX-2 mRNA expression, but results in higher degree of cellular and intracellular co-localization of their immunoreactive proteins in human placenta trophoblast cells.
PGE2 is involved in initiation and progression of labor in many species. Biosynthesis of PGE2 is mediated by cyclooxygenases (COX) and prostaglandin E synthases (PGES). mPGES-1 and COX-2 form an inducible pathway for PGE2 production in many cell systems. In this study we investigated whether mPGES-1 is involved in cytokine induced PGE2 biosynthesis in human trophoblast cells. We have evaluated the cellular and intracellular co-localization of mPGES-1 and COX-2, as well as cPGES and COX-1 in human trophoblast cells by dual immunofluorescent staining. The effect of IL-1beta on mPGES-1 and COX-2 co-localization, such as would occur with infection, and the regulatory effects of pro-inflammatory cytokines IL-1beta and TNF-alpha on transcriptional activity of mPGES-1 and COX-2 in these cells were also studied. We found that in cultured unstimulated trophoblasts, some cells expressed predominantly either mPGES-1 or COX-2, though there were cells co-expressing both enzymes. With IL-1beta treatment, mPGES-1 and COX-2 became more consistently co-localized. mPGES-1 was not transcriptionally co-induced with COX-2 by the cytokine treatment. We conclude that mPGES-1 is not involved in the inducible COX-2 mediated pathway for PGE2 biosynthesis at the transcriptional level, however, the treatment with IL-1beta results in a higher degree of co-ordination of the mPGES-1 and COX-2 protein immunolocalization, eliciting PGE2 synthesis.


Trisomy 12 and 4 in a thecoma of the ovary.
Cytogenetic analysis of short-term tissue culture from a thecoma of the ovary demonstrated the presence of trisomies of chromosomes 12 and 4 in all analyzed cells. Our finding confirms the consistency with which trisomy 12 is observed in benign sex cord/stromal tumors and suggests that trisomy 4 may be a second event in tumorigenesis of thecoma.


Possible use of Trichuris suis ova in autism spectrum disorders therapy.
Autism and autism spectrum disorders (ASDs) are heterogeneous, severe neurodevelopmental pathologies. The main core symptoms are: dysfunctions in social interactions and communication skills, restricted interests, repetitive and stereotypic verbal and non-verbal behaviors. Several biochemical processes are associated with ASDs: oxidative stress; endoplasmic reticulum stress; decreased methylation capacity; limited production of glutathione; mitochondrial dysfunction; intestinal dysbiosis; increased toxic metal burden; immune dysregulation. Current available treatments for ASDs can be divided into behavioral, nutritional and medical approaches, although no defined standard approach exists. Current drugs fail to benefit the ASD core symptoms and can have marked adverse effects, are mainly palliative and only sometimes efficacy in attenuating specific autistic behaviors. Helminthic therapy shows potential for application as anti-inflammatory agent. Several human diseases can be treated by helminths (i.e. inflammatory bowel disease, asthma, multiple sclerosis and autoimmune diabetes). Trichuris suis ova (TSO) show strong immunomodulatory properties. Authors hypothesize that TSO could be useful in addressing ASD immune dysregulations. TSO could be a novel therapeutic option for ASD management.


SELAVCO: a method to deal with trial-to-trial variability of evoked potentials.
The possibility and the potential dangers of the inhomogeneity of potentials to repeated stimuli have been recognized for some time. The predominant method of averaging sweeps of brain activity time locked to the stimuli relies on the assumption of an invariant response. In this paper, a method is proposed for dealing more effectively with the variability of single potentials than does averaging. This method operates by weighted averaging and the weights are determined from the data after suitable filtering operations. SELAVCO is validated and compared to the average for real data and in a controlled design close to reality.


Pattern of movement in and dispersal from a dutch forest of Carabus problematicus Hbst. (Coleoptera, Carabidae).
Data in the literature that in the Netherlands Carabus problematicus is an autumn breeder with overwintering larvae and adults have been confirmed. Pitfall catches show two peaks of adult beetles. The first small peak of activity is realized in May, the significance of which is not yet understood. The second higher peak occurs from the end of August till the beginning of October: the reproduction period. In this period at least three age classes are present. Some females were shown to take part in reproduction for at least two seasons. Old females appeared to have less eggs in the ovaries than young ones. Larvae can be trapped throughout the winter. The new generation appears in July and in the beginning of August. They immediately take part in reproduction. C. problematicus shows noctural activity with a peak towards the end of the night.In field experiments the pattern of movement and the dispersal behaviour of marked individuals was investigated and related to the choice of habitat. It was shown that in the Netherlands C. problematicus prefers a forest habitat. Some individuals were found to disperse from their favoured habitat. The percentage of beetles that disperse out of the forest during the breeding season could be estimated at 4-8%. Dispersal increases in the second part of the breeding season. Females tend to disperse more frequently than males.The pattern of movement consists of two types: a "random walk" and a "directed movement". It is suggested that the pattern of movement of individuals dispersing out of the forest is the directed movement. It could be shown that individuals released outside the forest were able to walk towards a forest silhouette. The significance of both the pattern of movement and the dispersal behaviour for the survival of the species is discuussed.


ASL Metabolically Regulates Tyrosine Hydroxylase in the Nucleus Locus Coeruleus.
Patients with germline mutations in the urea-cycle enzyme argininosuccinate lyase (ASL) are at risk for developing neurobehavioral and cognitive deficits. We find that ASL is prominently expressed in the nucleus locus coeruleus (LC), the central source of norepinephrine. Using natural history data, we show that individuals with ASL deficiency are at risk for developing attention deficits. By generating LC-ASL-conditional knockout (cKO) mice, we further demonstrate altered response to stressful stimuli with increased seizure reactivity in LC-ASL-cKO mice. Depletion of ASL in LC neurons leads to reduced amount and activity of tyrosine hydroxylase (TH) and to decreased catecholamines synthesis, due to decreased nitric oxide (NO) signaling. NO donors normalize catecholamine levels in the LC, seizure sensitivity, and the stress response in LC-ASL-cKO mice. Our data emphasize ASL importance for the metabolic regulation of LC function with translational relevance for ASL deficiency (ASLD) patients as well as for LC-related pathologies.


Myeloid differentiation (MyD)/growth arrest DNA damage (GADD) genes in tumor suppression, immunity and inflammation.
Myeloid differentiation (MyD) primary response and growth arrest DNA damage (Gadd) genes comprise a set of overlapping genes, including known (IRF-1, EGR-1, Jun) and novel (MyD88, Gadd45alpha, MyD118/Gadd45beta, GADD45gamma, MyD116/ Gadd34) genes, that have been cloned by virtue of being co-ordinately induced upon the onset of terminal myeloid differentiation and following exposure of cells to stress stimuli. In recent years it has become evident that MyD/Gadd play a role in blood cell development, where they function as positive regulators of terminal differentiation, lineage-specific blood cell development and control of blood cell homeostasis, including growth inhibition and apoptosis. MyD/Gadd are also involved in inflammatory responses to invading micro-organisms, and response to environmental stress and physiological stress, such as hypoxia, which results in ischemic tissue damage. An intricate network of interactions among MyD/GADD genes and gene products appears to control their diverse functions. Deregulated growth, increased cell survival, compromised differentiation and deficiencies in DNA repair are hallmarks of malignancy and its progression. Thus, the role MyD/Gadd play in negative growth control, including cell cycle arrest and apoptosis, and in DNA repair, make them attractive molecular targets for tumor suppression. The role MyD/Gadd play in innate immunity and host response to hypoxia also make these genes and gene products attractive molecular targets to treat immunity and inflammation disorders, such as septic shock and ischemic tissue damage.