Cumulative incidence of SARS-CoV-2 infection in the general population of the Valencian Community (Spain) after the surge of the Omicron BA.1 variant.
Studies investigating the cumulative incidence of and immune status against SARS-CoV-2 infection provide valuable information for shaping public health decision-making. A cross-sectional study on 935 participants, conducted in the Valencian Community (VC), measuring anti-SARS-CoV-2-receptor binding domain-RBD-total antibodies and anti-Nucleocapsid (N)-IgGs via electrochemiluminescence assays. Quantitation of neutralizing antibodies (NtAb) against ancestral and Omicron BA.1 and BA.2 variants and enumeration of SARS-CoV-2-S specific-IFNγ-producing CD4+ and CD8+ T cells was performed in 100 and 137 participants, respectively. The weighted cumulative incidence was 51.9% (95% confidence interval [CI]: 48.7-55.1) and was inversely related to age. Anti-RBD total antibodies were detected in 97% of participants; vaccinated and SARS-CoV-2-experienced (VAC-ex; n = 442) presented higher levels (p < 0.001) than vaccinated/naïve (VAC-n; n = 472) and nonvaccinated/experienced (UNVAC-ex; n = 63) subjects. Antibody levels correlated inversely with time elapsed since last vaccine dose in VAC-n (Rho, -0.52; p < 0.001) but not in VAC-ex (rho -0.02; p = 0.57). Heterologous booster shots resulted in increased anti-RBD antibody levels compared with homologous schedules in VAC-n, but not in VAC-ex. NtAbs against Omicron BA.1 were detected in 94%, 75%, and 50% of VAC-ex, VAC-n and UNVAC-ex groups, respectively. For Omicron BA.2, the figures were 97%, 84%, and 40%, respectively. SARS-CoV-2-S-reactive IFN-γ T cells were detected in 73%, 75%, and 64% of VAC-ex, VAC-n and UNVAC-ex, respectively. Median frequencies for both T-cell subsets were comparable across groups. In summary, by April 2022, around half of the VC population had been infected with SARS-CoV-2 and, due to extensive vaccination, displayed hybrid immunity.


Introduction of definite amounts of nonpermeant molecules into living cells after electropermeabilization: direct access to the cytosol.
The possibility of introducing definite amounts of nonpermeant molecules into electropermeabilized living cells has been approached by quantifying the amounts of Lucifer Yellow (LY; a 457-Da highly fluorescent molecule) and Phytolacca americana (Pokeweed) antiviral protein (PAP; a 30,000-Da ribosome-inactivating protein) retained by the cells after closure of the electric-field-induced transient structures of permeation. Without the electropermeabilization, these two molecules enter the cell only in very small amounts by fluid-phase pinocytosis. Under our experimental conditions, using the NIH 3T3 cells, the intracellular LY concentration can reach a value equivalent to the extracellular concentration and can be regulated by controlling the external concentration. We describe the use of LY in a rapid and efficient test for the determination of the best electrical-shock conditions of other cell lines. After electropermeabilization, PAP is 2 X 10(5) times more cytotoxic. Its toxicity can be detected at external concentrations (10(-11) M) corresponding to less than 10 internalized molecules per electropermeabilized cell. Therefore, after electropermeabilization, the nonpermeant molecules have a direct access to the cytosol and the biological effect of nonpermeant substances can be revealed.


Liquid-based cytology versus conventional Papanicolaou smear in an organized screening program : a prospective randomized study.
The objective of this study was to evaluate whether liquid-based cytology (LBC) can improve high-standard cervical cancer screening cytology further. The primary endpoint was histopathologic high-grade lesions in current and subsequent screening rounds. The secondary endpoints were cytologic diagnosis and inadequate samples. Women were randomized to smear taking by conventional Papanicolaou (Pap) smear or LBC according to the time of appointment. Eight thousand eight hundred ten conventional Pap smears and 4674 LBC samples were included. Evaluations of atypical cytology and referral to colposcopy and treatment were performed as routine procedures. Histopathologic diagnoses were retrieved from a regional database 8 months after the study was closed. The mean follow-up was 2 years and 9 months. Inadequate samples were observed in 0.3% of LBC samples versus 0.7% of Pap smears (P = .002). The total fraction of nonbenign diagnoses in cytology was 4.5% versus 3.5%, respectively (P < .001). Histopathologic evaluation was made on 570 patients constituting 4.6% of the LBC samples and 4% of the Pap smears. Forty percent more high-grade lesions were identified as a result of LBC sampling (1.20% vs 0.85%; P = .05). The influence of the sampling method was significant for all variables (odds ratio [OR], 1.60; 95% confidence interval [95% CI], 1.12-2.28) for high-grade lesions that were identified by histology when adjusting for age and screening unit in a logistic regression model. At the second follow-up 2 years and 1 month later, the OR was decreased only slightly (1.51; 95% CI, 1.13-2.01). In the ongoing cervical screening program of western Sweden, liquid cytology produced a significantly higher yield of histologic high-grade lesions compared with conventional Pap smears.


A review on potential sulfide-based ternary chalcogenides for emerging photo-assisted water purification applications.
Sulfide-based ternary chalcogenides have been recognized widely as exceptional photocatalysts, thanks to their narrow band gap enabling them to harvest solar energy to the maximum extent. They provide excellent optical, electrical, and catalytic performance and are of abundant use as a heterogeneous catalyst. Among sulfide-based ternary chalcogenides, compounds exhibiting AB2X4 structure form a new class of materials with excellent stability in photocatalytic performance. In the AB2X4 family of compounds, ZnIn2S4 is one of the top performing photocatalyst for energy and environmental applications. However, to date, only limited information is available on the mechanism behind the photo-induced migration of charge carriers in ternary sulfide chalcogenides. Ternary sulfide chalcogenides with their visible region activity and substantial chemical stability greatly depend on crystal structure, morphology, and optical characteristics for their photocatalytic activity. Hence, in this review, a comprehensive assessment of the reported strategies for enhancement of the photocatalytic efficiency of this compound is presented. In addition, a meticulous investigation of the applicability of ternary sulfide chalcogenide compound ZnIn2S4, in particular, has been delivered. Also, the photocatalytic behavior of other sulfide-based ternary chalcogenides for water remediation applications has also been briefed. Finally, we conclude with an insight into the challenges and future advancements in the exploration of ZnIn2S4-based chalcogenide as a photocatalyst for various photo-responsive applications. It is believed that this review could contribute to a better understanding of ternary chalcogenide semiconductor photocatalysts for solar-driven water treatment applications.


Raf kinase inhibitor protein (RKIP) and phospho-RKIP expression in melanomas.
Melanoma, a cancer notorious for its high potential to metastasize, arises from melanocytes, cells dedicated to melanin production and located in the basal layer of the epidermis. Raf-1 kinase inhibitor protein (RKIP) is an inhibitory molecule that down-regulates the effects of the Ras/Raf/MEK/ERK signaling pathway. The aim of this study was to examine the expression of RKIP and pRKIP in melanomas at different stages. We evaluated the RKIP and pRKIP protein by immunohistochemistry in control skin, pigmented nevi and melanomas, and through Western blotting in human normal melanocytes and in four different melanoma-derived cell lines (WM35, A375, M14, and A2058). Our results demonstrated a correlation between the expression of RKIP and pRKIP, and metastatic ability in melanoma cells. This raises the possibility to analyze both RKIP and pRKIP in all melanomas. Down-regulation of both RKIP and pRKIP expression could represent a useful marker of metastatic melanoma. On the contrary for non-metastatic melanoma, especially in Clark I and II, low RKIP and high pRKIP expression could be indicative. In conclusion, the observed negative correlation of the RKIP and pRKIP expression in metastatic melanomas indicates that expression of these proteins may become a prognostic marker for the progression of human cutaneous melanoma. We propose that the investigation of both RKIP and pRKIP may provide a useful tool indicative for metastatic or non-metastatic melanoma in different Clark's level melanomas. Further studies are required to verify the molecular background of the observed RKIP and pRKIP variations.


Solution NMR of membrane proteins in bilayer mimics: small is beautiful, but sometimes bigger is better.
Considerable progress has been made recently on solution NMR studies of multi-transmembrane helix membrane protein systems of increasing size. Careful correlation of structure with function has validated the physiological relevance of these studies in detergent micelles. However, larger micelle and bicelle systems are sometimes required to stabilize the active forms of dynamic membrane proteins, such as the bacterial small multidrug resistance transporters. Even in these systems with aggregate molecular weights well over 100 kDa, solution NMR structural studies are feasible-but challenging.


Quantification of atrazine and its metabolites in urine by on-line solid-phase extraction-high-performance liquid chromatography-tandem mass spectrometry.
We have developed a method using on-line solid-phase extraction-high-performance liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS) and isotope dilution quantification to measure atrazine and seven atrazine metabolites in urine. The metabolites measured were hydroxyatrazine, diaminochloroatrazine, desisopropylatrazine, desethylatrazine, desethylatrazine mercapturate, atrazine mercaturate and atrazine itself. Our method has good precision (relative standard deviations ranging from 4 to 20% at 5, 10 and 50 ng/mL), extraction efficiencies of 67 to 102% at 5 and 25 ng/mL, relative recoveries of 87 to 112% at 5, 25, 50 and 100 ng/mL limits of detection (LOD) ranging from 0.03 to 2.80 ng/mL. The linear range of our method spans from the analyte LOD to 100 ng/mL (40 ng/mL for atrazine and atrazine mercapturate) with R (2) values of greater than 0.999 and errors about the slope of less than 3%. Our method is rapid, cost-effective and suitable for large-scale sample analyses and is easily adaptable to other biological matrices. More importantly, this method will allow us to better assess human exposure to atrazine-related chemicals.


Pseudorabies virus glycoprotein III derived from virions and infected cells binds to the third component of complement.
Glycoprotein III (gIII) of pseudorabies virus (PRV) was shown to bind to the third component of complement (C3). This was observed only with porcine C3 whereas human C3 showed negligible binding under the conditions tested. PRV virion proteins could be precipitated from supernatants and cell lysates of PRV-infected cells by means of swine-C3 coupled to sepharose. According to their molecular size and their reactivity with anti-gIII monoclonal antibodies, the precipitated PRV proteins represented the fully glycosylated and smaller forms of the gIII protein. Precipitation from PRV virions yielded predominantly the fully glycosylated form of gIII whereas infected cell lysates also contained lower molecular weight gIII proteins. The observed specificity of the virus protein for porcine C3 correlates well with the known host tropism of PRV. Our findings suggest that PRV gIII may exhibit more functions than solely providing attachment to heparin-like moieties on target cell surfaces. As the complement cascade is an important defense mechanism against a variety of pathogens, the interaction with the host C3, the pivotal component of the complement activation, might be a virulence factor of PRV.


Response of hemopoietic cells to avian acute leukemia viruses: effects on the differentiation of the target cells.
Chicken bone marrow cells were infected with three avian acute leukemia viruses (ALV)--avian myeloblastosis virus (AMV), myelocytomatosis virus strain MC29 and Mill Hill 2 virus (MH2)--and then cultured in agar in the presence of conditioned medium. Under these conditions, it was found that very few cells served as target cells for these three viruses. Density gradient separation showed that ALV target cells were found primarily in the light density fractions and might be represented by cells committed to the mononuclear phagocyte pathway. Separation of bone marrow cells on the basis of their sedimentation velocity at unit gravity suggested that MC29 and AMV did not share the same target cells. In addition, the analysis of surface receptors and functional markers characteristic of macrophages (Fc and complement receptors, phagocytosis and immune phagocytosis) indicated that the ALV-transformed cells were blocked during their differentiation. These results indicate that the transforming ability of ALV interferes with the differentiation of their target cells.


Diagnosis and therapy of renal tubular acidosis in infancy.
While distrubances of the acid-base balance are frequently seen in infancy, renal tubular acidosis is a rather rare disease but should be considered as differential diagnosis if metabolic acidosis persists after adequate treatment. Proximal and distal tubular acidosis with primary and secondary forms can be differentiated. Proximal RTA is characterized by the loss of bicarbonate, distal RTA by a defect to establish a hydrogen ion gradient and thus to accomplish acidification of urine. In addition to these two basic forms a bicarbonate wasting state in distal RTA has been described. A patient with these clinical features is presented. He was admitted to our hospital at the age of 1 month with meningitis, enteritis and marked dystrophy. A persistant hyperchloraemic acidosis with concomitant hypokalaemia was present. The ammonium chloride loading test confirmed the diagnosis of primary distal RTA. Renal biopsy performed with 1 year of age revealed nephrocalcinosis of the inner medullary region of the kidney while the cortex was not affected. The patient first needed alkali doses of 12 mEq/kg/day which could be gradually reduced to 3.5 mEq/kg/day. Under additional potassium substitution of 5 mEq/kg/day he was thriving well. Differential diagnosis and the particular clinical features of this case are discussed.


To Work or Not to Work Remotely? Work-to-family Interface Before and During the COVID-19 Pandemic.
This paper provides a brief, evidence-based reflection on the differences between 'old-normal' remote working and mandatory work-from-home during the Covid-19 pandemic. From the perspective of applied psychology in the field of work and organizations, we used self-report instruments to assess variations in work-family conflict and enrichment, frequency of information and communication technologies use, and recovery in two longitudinal studies. The first study involved 148 individuals from the technical-administrative staff of a large Italian University, during an experimentation of remote working (one day per week) in 2019. The second study, conducted during the first lockdown in 2020, involved 144 individuals (occasional sample, heterogeneous by profession). All participants completed a self-report online questionnaire two times six months apart. Although the two studies are not directly comparable, the results showed two different situations: in the condition of experimental remote working (one day per week), participants reported a decrease in work-family conflict and an improvement in recovery experiences, while in the emergency remote working condition, a deterioration in work-family enrichment was found. These findings highlighted that some differences exist between a planned remote working condition and a mandatory one in an emergency. We briefly discuss these aspects to inform future organizational decisions and actions for the 'new normal'.


Impact of a high-fat diet containing canola or soybean oil on body development and bone; parameters in adult male rats.
The role of the fatty acid in the prevention or progression of chronic diseases has generated significant interest on the part of researchers. Thus, our objective was to evaluate the long-term effects of high-fat diet containing soybean or canola oil on body development and bone parameters of male rats. After weaning, rats were grouped and fed either a control diet (7S) or a high-fat diet containing soybean (19S) or canola oil (19C). Femur and lumbar vertebra (LV4) structure were determined at 180 days by dual-energy X-ray absorptiometry and computed tomography. The groups showed similar food intake, body mass and length development. The bone parameters of the 19C were similar to the control group, while the 19S showed lower bone parameters when compared to the other groups. The high-fat diet containing soybean oil was unfavorable to bone structure, while the canola oil contributed bone health during the adult stage of life.


Vitamin D Deficiency in Critically Ill Children with Sepsis: What is the Road ahead?
Sivaraman RP. Vitamin D Deficiency in Critically Ill Children with Sepsis: What is the Road ahead? Indian J Crit Care Med 2021;25(8):843-844.


Reproductive responses of quail to bunker C oil fractions.
Bunker C fuel oil was extracted with petroleum ether and the concentrated extract was given to breeding Japanese quail (Coturnix coturnix Japonica). A dose of extract equivalent to 300 mg bunker C reduced both egg production and hatchability. Fractionation of the extract by absorption on Florisil and elution with petroleum ether followed by chloroform yielded fractions that were tested for toxicity. All the fractions had a pronounced effect upon production; however, only the chloroform eluates were found to affect hatchability of the eggs.


An audit of magnetic resonance imaging in the paediatric orthopaedic setting.
To determine when magnetic resonance imaging (MRI) is helpful in the management of a selected group of paediatric orthopaedic patients. A retrospective analysis of 131 MRI scans was undertaken with allocation into seven categories based on clinical presentation. MRIs performed for spinal, congenital and intra-articular pathology, as well as for growth plate assessment correlated well with subsequent clinical and/or surgical findings. Three of ten MRIs (30%) incorrectly assessed whether a foreign body was present with a sensitivity of 0.60 and specificity of 0.80. Three of 20 MRIs (15%) could not accurately distinguish between oedema/effusion and frank infection. Here sensitivity was 1.00 and specificity was 0.73. MRIs performed for assessment of tumours were accurate with respect to margins and extent. Assessment of pathology with MRI was never the indication for MRI, thus it was not surprising that in four of 27 soft tissue tumours (sensitivity of 0.88 and specificity of 0.63) MRI did not correlate with subsequent histological findings. MRI plays an important role in the assessment of a wide range of musculo-skeletal pathology. MRI does not, and could not be expected to, replace the need for incisional biopsy for tumour diagnosis. It must be used with caution where tissues have been previously explored for foreign bodies. Interpretation of MRI in musculoskeletal infection must consider its timing in the evolution of the patient's infection.


Development of Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) in bryozoan hosts (as examined by light microscopy) and quantitation of infective dose to rainbow trout (Oncorhynchus mykiss).
The myxozoan parasite Tetracapsuloides bryosalmonae is the causative agent of proliferative kidney disease (PKD), a highly damaging disease of cultured salmonid fish. Within this study, phylactolaemate bryozoans were collected from a river known to be endemic for PKD and subsequently cultured in the laboratory. Sequential developmental stages of T. bryosalmonae were studied by light microscopy within the living bryozoan colonies, allowing the identification of stages attached to host peritoneum, consistent with previous molecular evidence of cryptic stages. Infection resulted in the production of large numbers of spores, which were released from the bryozoans. Experimental exposure of rainbow trout (Oncorhynchus mykiss) to medium in which infected bryozoans were cultured resulted in clinical PKD. Rainbow trout were exposed to known numbers of T. bryosalmonae spores collected by micromanipulation, which had been released from mature spore sacs within colonies of the bryozoan Fredericella sultana. Exposure to one spore was sufficient to lead to development of PKD. These findings indicate that small numbers of bryozoans are capable of releasing sufficient spores to infect large numbers of fish, having implications for future control methods for PKD in salmonid farming.


Activation of liver X receptors inhibits pancreatic islet beta cell proliferation through cell cycle arrest.
Liver X receptors (LXRs) are important transcriptional regulators of lipid homeostasis and proliferation in several cell types. However, the roles of LXRs in pancreatic beta cells have not been fully established. The aim of this study was to investigate the effects of LXRs on pancreatic beta cell proliferation. Gene expression was analysed using real-time RT-PCR. Transient transfection and reporter gene assays were used to determine the transcriptional activity of LXRs in pancreatic beta cells. Cell viability and proliferation were analysed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), DNA fluorometric, BrdU labelling and [(3)H]thymidine incorporation assays. Cell cycle distribution was investigated by flow cytometry analysis. Adenovirus-based RNA interference was used to knockdown LXRalpha, LXRbeta and p27 in MIN6 cells and mouse islets. We found that both Lxralpha (also known as Nr1h3) and Lxrbeta (also known as Nr1h2) were expressed and transactivated the LXR response element in HIT-T15 and MIN6 cells. Activation of LXRs dose-dependently inhibited pancreatic beta cell viability and proliferation. This was accompanied by beta cell cycle arrest at the G1 phase. Furthermore, LXR activation increased levels of the p27 protein by inhibiting its degradation. Knockdown of p27 reversed these effects of LXR activation on growth inhibition and cell cycle arrest. Our observations indicate that LXR activation inhibits pancreatic beta cell proliferation through cell cycle arrest. A well-known regulator of pancreatic beta cell cycle progression, p27, is upregulated and mediates the effects of LXRs on growth inhibition in beta cells. These observations suggest the involvement of aberrant activation of LXR in beta cell mass inadequacy, which is an important step in the development of type 2 diabetes.


Non-enzymatic electrochemical glucose sensor based on platinum nanoflowers supported on graphene oxide.
A non-enzymatic electrochemical method was developed for glucose detection using a glassy carbon electrode modified with platinum nanoflowers supported on graphene oxide (PtNFs-GO). PtNFs-GO was synthesized using a nontoxic, rapid, one-pot and template-free method. Low-cost, green solvent ethanol acted as the reductant, and the advanced and effective 2D carbon material-GO nanosheet acted as the stabilizing material. Their morphologies were characterized using transmission electron microscopy. Cyclic voltammetry and amperometric methods were used to evaluate the electrocatalytic activity towards glucose in neutral media. The modified electrode exhibited strong and sensitive amperometric responses to glucose even in the presence of a high concentration of chloride ions. The response time was within 5s. The interference effects from ascorbic acid and uric acid were comparatively small when operated at suitable potential. Under optimal detection potential (0.47 V with a saturated calomel reference electrode) the PtNFs-GO modified electrode performed a current response towards glucose at a broad concentration range from 2 μM to 20.3mM. Two linear regions could be observed at 2 μM to 10.3mM with a sensitivity of 1.26 μA mM(-1)cm(-2) (correlation coefficient 0.9968), and at 10.3mM to 20.3mM with a sensitivity of 0.64 μA mM(-1)cm(-2)(correlation coefficient 0.9969). The LOD of 2 μM was lower than many non-enzymatic electrochemical glucose sensors. The modified electrode was also applied to the determination of glucose in glucose injection solutions, and the satisfactory results obtained indicated that it was promising for the development of a novel non-enzymatic electrochemical glucose sensor.


Metal-mediated Fe3O4@polydopamine-aptamer capture nanoprobe coupling multifunctional MXene@Au@Pt nanozyme for direct and portable photothermal analysis of circulating breast cancer cells.
Breast cancer (BC) is the most common cancer in the world and circulating tumor cells (CTCs) are reliable biomarkers for early breast cancer diagnosis in a non-invasive manner. However, effective isolation and sensitive detection of BC-CTCs by portable devices in human blood samples are extremely challenging. Herein, we proposed a highly sensitive and portable photothermal cytosensor for direct capture and quantification of BC-CTCs. To achieve efficient isolation of BC-CTCs, aptamer functionalized Fe3O4@PDA nanoprobe was facilely prepared through Ca2+-mediated DNA adsorption. To further detect the captured BC-CTCs with high sensitivity, multifunctional two-dimensional Ti3C2@Au@Pt nanozyme was synthesized, which not only possessed superior photothermal effect but also exhibited high peroxidase-like activity for catalyzing 3,3',5,5'-tetramethylbenzidine (TMB) to produce TMB oxide (oxTMB) with a strong photothermal characteristic, combining with Ti3C2@Au@Pt to synergistically amplify the temperature signal. Moreover, numerous Ti3C2@Au@Pt nanocomposites would be selectively attained on the BC-CTCs surface through multi-aptamer recognition and binding strategy, which further enhanced the specificity and facilitated signal amplification. Therefore, direct separation and highly sensitive detection of BC-CTCs was successfully achieved in human blood samples. More significantly, the controlled release of the captured BC-CTCs without affecting cell viability could be straightforwardly realized by a simple strand displacement reaction. Thus, with the distinct features of portability, high sensitivity, and easy operation, the current method holds great promise for early diagnosis of breast cancer.


Potassium Channel Remodeling in Heart Disease.
Heart disease produces substantial remodeling of K(+) channels that in general promotes arrhythmia occurrence. In the case of ventricular arrhythmias, K(+) channel remodeling contributes to the arrhythmic risk and increases vulnerability to torsades de pointes with K(+) channel inhibiting drugs. Atrial K(+) channel remodeling caused by atrial fibrillation promotes arrhythmia stability and presents opportunities for the development of new drugs targeting atrial inward rectifier K(+) currents. A better understanding of K(+) channel remodeling will help clinicians to appreciate arrhythmia mechanisms and determinants in a variety of clinical situations and to better manage arrhythmia therapy in patients with heart disease.


Transcending the impenetrable: how proteins come to terms with membranes.
In the living cell, proteins are efficiently sorted to a whole range of subcellular compartments. In many cases, sorting specificity is mediated by short 'sorting signals' attached either permanently or transiently to the protein. At long last, a fairly coherent picture of the design and function of many such sorting signals is beginning to emerge.


Gastrointestinal permeability in patients with irritable bowel syndrome assessed using a four probe permeability solution.
Abnormal gastrointestinal permeability has been linked to irritable bowel syndrome (IBS). The lactulose-to-mannitol ratio is traditionally used to assess small intestine permeability while sucralose and sucrose are used to assess colonic and gastric permeability respectively. We used a single 4-probe test solution to assess permeability throughout the gastrointestinal tract in IBS patients and healthy controls by measuring the recovery of the probes in urine after ingestion using a modified liquid chromatography mass spectrometry protocol. Fasting participants (N=59) drank a permeability test solution (100ml: sucralose, sucrose, mannitol, and lactulose). Urine was collected over a 5-h period and kept frozen until analysis. Urinary sugar concentrations were measured using a liquid chromatography/triple quadruple mass spectrometer. Colonic permeability was significantly lower in IBS patients when compared to healthy controls (p=0.011). Gastric and small intestinal permeability did not significantly differ between the groups. The study demonstrates the clinical potential of this non-invasive method for assessing alterations in gastrointestinal permeability in patients with IBS.


Mapping remodeling of thalamocortical projections in the living reeler mouse brain by diffusion tractography.
A major challenge in neuroscience is to accurately decipher in vivo the entire brain circuitry (connectome) at a microscopic level. Currently, the only methodology providing a global noninvasive window into structural brain connectivity is diffusion tractography. The extent to which the reconstructed pathways reflect realistic neuronal networks depends, however, on data acquisition and postprocessing factors. Through a unique combination of approaches, we designed and evaluated herein a framework for reliable fiber tracking and mapping of the living mouse brain connectome. One important wiring scheme, connecting gray matter regions and passing fiber-crossing areas, was closely examined: the lemniscal thalamocortical (TC) pathway. We quantitatively validated the TC projections inferred from in vivo tractography with correlative histological axonal tracing in the same wild-type and reeler mutant mice. We demonstrated noninvasively that changes in patterning of the cortical sheet, such as highly disorganized cortical lamination in reeler, led to spectacular compensatory remodeling of the TC pathway.


[Quantitative-morphological studies of the heart of MINI-LEWE minature swine. 2: Atrioventricular and semilunar valves].
Reference is made to the possible use of cardiac valves of animal origin as heteroplastic material in cardiac valve surgery. In that context, quantitative studies were conducted into all four cardiac valves. The data measured from clinically intact animals exhibited wide scatter. Different numbers of leaflets (between two and six) were recorded from atrioventricular valves, whereas three valvulae were always found on the semilunar valves. The volume of the tricuspid valve was significantly higher than that of the mitral valve, and among the semilunar valves bigger size was established for the aortic valve. Valve size grew along with body weight, as established by correlative analysis. The data measured by the examiners were compared with variables reported in literature.


Investigation of magnetic field effects on the dose-response of 3D dosimeters for magnetic resonance - image guided radiation therapy applications.
The strong magnetic field of integrated magnetic resonance imaging (MRI) and radiation treatment systems influences secondary electrons resulting in changes in dose deposition in three dimensions. To fill the need for volumetric dose quality assurance, we investigated the effects of strong magnetic fields on 3D dosimeters for MR-image-guided radiation therapy (MR-IGRT) applications. There are currently three main categories of 3D dosimeters, and the following were used in this study: radiochromic plastic (PRESAGE®), radiochromic gel (FOX), and polymer gel (BANG™). For the purposes of batch consistency, an electromagnet was used for same-day irradiations with and without a strong magnetic field (B0, 1.5T for PRESAGE® and FOX and 1.0T for BANG™). For PRESAGE®, the percent difference in optical signal with and without B0 was 1.5% at the spectral peak of 632nm. For FOX, the optical signal percent difference was 1.6% at 440nm and 0.5% at 585nm. For BANG™, the percent difference in R2 MR signal was 0.7%. The percent differences in responses with and without strong magnetic fields were minimal for all three 3D dosimeter systems. These 3D dosimeters therefore can be applied to MR-IGRT without requiring a correction factor.


Efficacy assessment and complications of surgical management for superior semicircular canal dehiscence: a meta-analysis of published interventional studies.
Superior semicircular canal syndrome (SSCS) includes vestibular and audiological symptoms which result from the introduction of a third mobile window into the osseous cochlea. Surgical repair is considered in cases of incapacitating symptoms. The present paper aims at comparing the different surgical approaches and modes of dehiscence repair, regarding their respective efficacy and potential pitfalls. A systematic literature review and meta-analysis of pooled data were performed. Study selection included prospective- and retrospective-controlled studies, prospective- and retrospective-cohort studies, ex vivo studies, animal models, case-reports, systematic reviews and clinical guidelines. A total of 64 primary operations for SSC repair were identified; 56 ears were operated for vestibular and 7 for auditory complaints. A total of 33 ears underwent canal plugging, 16 resurfacing, and 15 capping. Success rates were 32/33, 8/16, and 14/15, respectively. The observed differences were statistically significant (P=0.001). Resurfacing proved less effective than both plugging (P=0.002), and capping (P=0.01) techniques. Temporalis fascia was commonly used as sealing material and was combined with bone-pâté/bone-wax (plugging), bone-graft (resurfacing), or hydroxyapatite-cement (capping). Most operations were performed via middle-fossa approach; higher success rates were associated with plugging and capping techniques. SNHL and disequilibrium were the most frequent complications encountered. Most cases were followed for 3-6 months. Precise criteria regarding follow-up duration and objective success measures are not determined. Surgical repair of SSCS is considered as a valid therapeutic option for patients with debilitating symptoms. Consensus regarding strict follow-up criteria and objective assessment of success is necessary before larger scale operations can be implemented in clinical practice.


Cross infections due to coagulase-negative staphylococci in high-risk patients.
Until recently, infections due to coagulase-negative staphylococci (CNS) have been regarded as endogenous in origin. However, there are now increasingly reports in the literature on the endemic occurrence of distinct strains of CNS. Several outbreaks due to CNS are reported in cardiac surgery or in neonates. The latter seem to be high risk populations in regard to CNS infections because of certain risk factors (i.e. degree of immunosupression, routine use of central venous catheters and parenteral lipids as well as broad spectrum antibiotic therapy). On the other hand, these newborn babies have no physiological skin flora and are therefore easily colonized by multiresistent bacteria. The persistence of certain well-defined Staphylococcus epidermidis (SE) strains in neonatal intensive care units have been demonstrated over periods as long as a decade. Specific putative virulence factors (i.e. slime production and polysaccharide/adhesin PS/A) were more common in endemic strains as compared to single isolates. Pulsed-field gel electrophoresis (PFGE) proves to be a powerful tool in the study of the epidemiology of CNS while other modern typing techniques (ribotyping, plasmid typing) were also used in the literature to investigate outbreaks of CNS infections.


Integrating POLE/POLD1 mutated for immunotherapy treatment planning of advanced stage non-small cell lung cancer.
In this study, we evaluated the potential of DNA polymerase epsilon (POLE) and DNA polymerase delta 1 (POLD1) as prognostic biomarkers for immune checkpoint inhibitor (ICI) treatment in patients with advanced stage non-small cell lung cancer (NSCLC). Disease stage, PD-L1 positivity, histological subtypes, POLE/POLD1 mutation status, tumor mutation burden (TMB), and response to ICIs in NSCLC cases were derived from AACR GENIE dataset (n = 24 120), TCGA-Pan Lung Cancer dataset (n = 1144), AACR GENIE BPC NSCLC v2.0-public (n = 2004), and Memorial Sloan Kettering-Integrated Mutation Profiling of Actionable Cancer Targets dataset (n = 350). The smoking history from TCGA and AACR GENIE datasets was grouped into current, former or never-smokers. POLE and POLD1 genetic alterations were identified in 5% and 2.6% of NSCLC patients, respectively. Current smokers had 9% and 4% of POLE/POLD1 mutations, respectively, versus 1.7% for both POLE and POLD1 mutations prevalence in never-smokers. POLE/POLD1 mutations were associated with elevated mutation counts than those with wild-type (median mutation counts 16 vs. 7, p < 0.0001), more advanced disease stages (stage I disease 15.19% vs. 29.42%), more prevalent squamous histology subtype (21.69% vs. 9.05%, p = 0.0427), and a higher percentage of PD-L1 positivity (66.67% vs. 43.87%, p < 0.001). Treatment with ICIs improved survival in patients with both POLE/POLD1 mutated and those with TMB > 18 (p < 0.001). Current smokers have a five-fold increased risk of having POLE mutations than never-smokers. POLE/POLD1 mutation status and TMB > 18 can be a composite biomarker for selecting NSCLC patients with survival benefits to ICI treatment.


Co-chairman's remarks: genetic recombination in the molecular era.
The present status of some general questions about DNA recombination is assessed. Topics include the mechanisms of synapsis and strand exchange, and the functions of recombination in nature.


Parasitological and molecular detection of Trypanosoma evansi in a dog from Tocache, San Martin, Peru.
This study presents the first case report of canine trypanosomiasis caused by Trypanosoma evansi in Peru. The case was admitted to a veterinary clinic in the Peruvian Amazon region of San Martin with severe clinical symptomatology which resulted in the dog's death. Microscopy screening showed the presence of trypomastigotes in blood and bone marrow and postmortem histopathology found damage at the cardiac, lung, kidney and spleen levels. Collected specimens were tested by nested-PCR which were positive for Trypanosoma spp., but negative for T. cruzi. High-throughput sequencing determined that the infecting species was closely related to T. equiperdom/evansi and subsequent phylogenetic analysis confirmed that the sample was related to T. evansi. The presence of T. evansi in the area highlights the need for increased surveillance to assess the impact of surra in the region and to develop measures to prevent socioeconomic damage resulting from infections in domestic and farm animals as well as prevent zoonotic transmission.


Role of conventional chemosensitivity test and tissue biomarker expression in predicting response to treatment of peritoneal carcinomatosis from colon cancer.
Peritoneal carcinomatosis (PC) is observed in approximately 10% of patients with colorectal cancer at the time of primary cancer resection. Most of these patients receive 5-fluorouracil (5-FU)- or oxaliplatin-containing chemotherapy regimens as first-, second-, or third-line treatment. In the present study, sensitivity and resistance to drugs used to treat PC were better defined by a conventional chemosensitivity test than by biomarker expression. 5-Fluorouracil- or oxaliplatin-based regimens are the treatments of choice in patients with PC from colon cancer. There are currently no useful preclinical evaluations to guide the decision-making process for tailored therapy. The aim of the present study was to compare the advantages and limits of a conventional in vitro chemosensitivity test with those of a panel of biomolecular markers in predicting clinical response to different drugs used to treat colon cancer-derived PC. Fresh surgical biopsy specimens were obtained from 28 patients with peritoneal carcinomatosis from colon cancer. TS, TP, DPD, MDR1, MRP-1, MGMT, BRCA1, ERCC1, GSTP1, and XPD gene expression levels were determined by real-time reverse transcription polymerase chain reaction. An in vitro chemosensitivity test was used to define a sensitivity or resistance profile to the drugs used to treat each patient. Expression levels of the genes analyzed were generally poorly related to each other. TS and ERCC1 expression was inversely related to response to 5-FU-and/or oxaliplatin-containing regimens. Significant predictivity in terms of sensitivity but poor predictivity of resistance (56.2%) (P=.037) were observed for ERCC1 expression (90%), and high predictivity of resistance (100%) but very low predictivity of sensitivity (40%) (P=.014) were registered for TS. The best overall and significant predictivity was observed for chemosensitivity test results (62.5% sensitivity and 89% resistance; P=.005). Sensitivity and resistance to drugs used in vivo was better defined by the chemosensitivity test than by biomarker expression.


Prevalence of CCR5delta32 in Northeastern Iran.
A 32-base pair deletion (∆32) in the open reading frame (ORF) of C-C motif chemokine receptor 5 (CCR5) seems to be a protective variant against immune system diseases, especially human immunodeficiency virus type 1 (HIV-1). We aimed to assess the frequency of CCR5∆32 in the healthy Iranian population. In this study, 400 normal samples from Khorasan, northeastern Iran, were randomly selected. The frequency of CCR5∆32 carriers was investigated using PCR analysis. Allele prevalence and the fit to the Hardy-Weinberg equilibrium were analyzed. The prevalence of CCR5∆32 in the northeastern population of Iran was 0.016. Four hundred samples were studied, among which one with CCR5∆32/∆32 and 11 with CCR5Wild/∆32 genotype were detected. This study was the first investigation for an assessment of the prevalence of CCR5∆32 in northeastern Iran. The low prevalence of CCR5∆32 allele in the Iranian population may result in the increased susceptibility to HIV-1. In addition, this prevalence is the same as that of reported in East Asia, while is lower than that in the Europeans.


Enterocytozoon bieneusi (Microspora): prevalence and pathogenicity in AIDS patients.
Microsporidia are unicellular organisms, which lack mitochondria. They have prokaryotic-like ribosomes and characteristic spores containing an extrusible polar tube which serves as a passage for inoculation of the infectious agent (sporoplasm) into host cells. Clinically apparent infections in man appear to be limited to immunoprivileged sites or immunocompromised patients. One species, Encephalitozoon cuniculi, has been reported several times in patients with neurological disorders and once causing a fatal hepatitis in an AIDS patient. The most recently discovered species, Enterocytozoon bieneusi, is known only from the small intestinal enterocytes of patients with the acquired immunodeficiency syndrome, and is easily differentiated from other microsporidia by the precocious development of spore organelles in the sporont and by the poor development of the endospore layer of the spore wall. Although only about 40 cases have been reported, circumstantial evidence suggests that E. bieneusi may be the cause of a severe watery diarrhoea, which responds only temporarily to treatment with metronidazole.


Modeling thermal responses in human subjects following extended exposure to radiofrequency energy.
This study examines the use of a simple thermoregulatory model for the human body exposed to extended (45 minute) exposures to radiofrequency/microwave (RF/MW) energy at different frequencies (100, 450, 2450 MHz) and under different environmental conditions. The exposure levels were comparable to or above present limits for human exposure to RF energy. We adapted a compartmental model for the human thermoregulatory system developed by Hardy and Stolwijk, adding power to the torso skin, fat, and muscle compartments to simulate exposure to RF energy. The model uses values for parameters for "standard man" that were originally determined by Hardy and Stolwijk, with no additional adjustment. The model predicts changes in core and skin temperatures, sweat rate, and changes in skin blood flow as a result of RF energy exposure. The model yielded remarkably good quantitative agreement between predicted and measured changes in skin and core temperatures, and qualitative agreement between predicted and measured changes in skin blood flow. The model considerably underpredicted the measured sweat rates. The model, with previously determined parameter values, was successful in predicting major aspects of human thermoregulatory response to RF energy exposure over a wide frequency range, and at different environmental temperatures. The model was most successful in predicting changes in skin temperature, and it provides insights into the mechanisms by which the heat added to body by RF energy is dissipated to the environment. Several factors are discussed that may have contributed to the failure to account properly for sweat rate. Some features of the data, in particular heating of the legs and ankles during exposure at 100 MHz, would require a more complex model than that considered here.


Effects of dihydrotestosterone on brain mRNA levels of steroid 5alpha-reductase isozymes in early postnatal life of rat.
We studied the expression of type 1 (5alpha-R1) and type 2 (5alpha-R2) 5alpha-reductase isozymes (5alpha-R) and their regulation by dihydrotestosterone (DHT) in the prefrontal cortex of male and female rats during postnatal sexual differentiation of the central nervous system (CNS), using one-step quantitative RT-PCR coupled with laser-induced fluorescence capillary electrophoresis. We found a higher expression of 5alpha-R2, which is considered a masculinizing enzyme, in the female versus male CNS, and observed sexual dimorphism in the regulation of both 5alpha-R isozymes by DHT. These results open up a new research line that could improve understanding of the role of 5alpha-R isozymes in the physiology of the CNS.


Characterization of the novel HLA-DQA1*01:39 allele by next-generation sequencing.
DQA1*01:39 differs from DQA1*01:02:01:01 by one nucleotide substitution at position 49 in exon 1.


Evaluation of Sleep Habits and Disturbances Among US Adults, 2017-2020.
Unhealthy sleep behaviors and sleep disturbances are associated with higher risk of multiple diseases and mortality. The current profiles of sleep habits and disturbances, particularly the differences between workdays and free days, are unknown in the contemporary US. To comprehensively evaluate sleep habits on workdays and free days and the prevalence of sleep disturbances among US adults. This study is a cross-sectional analysis of US nationally representative data from the National Health and Nutrition Examination Survey (2017-2020) among adults aged 20 years or older. Data analysis was performed from February to May 2022. The main outcomes were means and/or distributions of sleep habits, including sleep duration and sleep-wake timing on workdays and free days, sleep debt (ie, the difference between sleep duration on free days and mean weekly sleep duration), and social jet lag (ie, the difference between the midpoint between sleep and wake time on workdays and free days). Prevalence of trouble sleeping (ie, participants told a doctor or other health professional that they have trouble sleeping) and daytime sleepiness (ie, self-reported feeling of being overly sleepy during the day ≥5 times per month) were also determined. A total of 9004 individuals (mean [SE] age, 48.3 [0.53] years; 4635 women [51.9%]; 3158 non-Hispanic White [62.8%]) were included in the current study. The mean sleep duration was 7.59 hours (95% CI, 7.54 to 7.64 hours) on workdays and 8.24 hours (95% CI, 8.17 to 8.31 hours) on free days (difference, 0.65 hour). The mean sleep and wake times were at 11:02 pm (95% CI, 10:57 pm to 11:17 pm) and 6:41 am (95% CI, 6:36 am to 6:45 am), respectively, on workdays and 11:25 pm (95% CI, 11:21 pm to 11:35 pm) and 7:41 am (95% CI, 7:37 am to 7:46 am), respectively, on free days (differences, 0.23 hour for sleep time and 1.00 hour for wake time). On workdays, 23.1% (95% CI, 21.3% to 24.9%) of adults slept less than 7 hours and 25.4% (95% CI, 24.1% to 26.6%) went to sleep at midnight or later; the corresponding percentages changed to 12.9% (95% CI, 11.6% to 14.1%) and 40.9% (95% CI, 38.4% to 43.5%), respectively, on free days. Furthermore, the mean sleep debt was 0.73 hours (95% CI, 0.68 to 0.77 hours), and mean social jet lag was 1.10 hours (95% CI, 1.05 to 1.15 hours); 30.5% (95% CI, 26.8% to 33.3%) of adults experienced 1 hour or more of sleep debt, and 46.5% (95% CI, 42.6% to 50.3%) experienced 1 hour or more of social jet lag. The prevalence of trouble sleeping was 29.8% (95% CI, 28.2% to 31.5%), and that of daytime sleepiness was 27.2% (95% CI, 25.0% to 29.5%). In 2017 to 2020, US adults showed variability in sleep habits between workdays and free days, with longer sleep duration and later sleep-wake phases on free days, and high percentages of US adults experienced long-term sleep deprivation, chronic social jet lag, and frequent sleep disturbances. These findings provide evidence to further investigate potential approaches to optimize overall US sleep health.


Bridging Stents in Fenestrated and Branched Endovascular Aneurysm Repair: A Systematic REVIEW.
Concern exists about durability of stent grafts used to bridge aortic grafts to visceral and renal arteries during fenestrated and branched endovascular aneurysm repair (F/B-EVAR). There are no guidelines regarding the ideal technique for joining target vessels (TVs). We systematically reviewed data published from 2014 to 2019 using PRISMA guidelines and PICO models. Keywords were searched in MEDLINE, EMBASE, and Cochrane Library. All articles were screened by two authors (a third author in case of discrepancies). Only original articles regarding F/B-EVAR in complex aortic aneurysm, reporting the number and type of TVs mated, the onset of bridging stent complications, and reinterventions on TVs were included. Analysis included quality assessment scoring, types of stent grafts, and complications related to bridging stents. 19 studies were included with 2,796 patients and 9556 TV; 4,797 renal arteries (50.2%), 4,174 visceral arteries (43.6%), and undefined TV (n = 585; 6.1%) were bridged. Balloon-expandable stent-grafts (B-EXP) were used in 40.9% and self-expandable (S-EXP) in 22.7% and undefined stents in 36.3%. The included studies had quality assessment scores ranging between 11/15 and 15/15, with high grade of accordance on reporting general results, but a low grade of accordance on reporting detailed data. Despite study heterogeneity, high-volume analysis confirmed a higher rate of complication in renal arteries than visceral arteries, 6% (95% CI 4-8) vs. 2% (95% CI 1-3), respectively. The rate of reinterventions was similar, 3% (95% CI 2-4) and 2% (95% CI 1-3). S-EXP versus B-EXP stent complication was 4% (95% CI 2-7) vs. 3% (95% CI 2-5), respectively. This systematic review underlines the low grade of accordance in reporting detailed data of bridging stents in F/B-EVAR. Renal TVs were more prone to complications, with an equivalent reintervention rate to visceral TVs. As to B-EVAR, the choice of B-EXP over S-EXP is still uncertain.


Strategies for musculoskeletal magnetic resonance imaging.
There is a broad range of clinical applications for MRI in the diagnosis of musculoskeletal disease, and the emphasis in this article is on the evaluation of the joints and extremities. A discussion of basic principles, including radiofrequency coils, positioning, pulsing sequences, imaging planes, and technical parameters is followed by discussions of tailored approaches to the evaluation of mass lesions and joint pathology for specific regions of the musculoskeletal system. In the final section, consideration is given to the overall role of MRI in the diagnosis of musculoskeletal disorders with respect to other established imaging modalities.


Teaching medical informatics to medical students in the Federal Republic of Germany.
The medical curriculum in the Federal Republic of Germany (FRG) is regulated by federal legislation. Medical informatics is part of this curriculum and is therefore taught by all 27 medical faculties in the FRG, 24 of which have related departments. The teaching situation in general and the specific approach at the author's department are described.


The Prognostic Value of TTF-1/NKX2-1 in Lung Squamous Cell Carcinoma.
TTF-1/NKX2-1 is a lineage-specific transcription factor that is expressed in the thyroid gland, lung, and forehead. It functions as a key component in regulating lung morphogenesis and differentiation. It is mainly expressed in lung adenocarcinoma, while its prognostic value in non-small-cell lung cancer remains controversial. This study evaluates the prognostic value of TTF-1 in different cellular locations in lung squamous cell carcinoma (SCC) and adenocarcinoma (ADC). The expression of TTF-1 was analyzed by immunohistochemistry in 492 patients (ADC 340 and SCC 152) who had undergone surgery between June 2004 and June 2012. Disease-free survival (DFS) and overall survival (OS) were estimated using the Kaplan-Meier method. Positive TTF-1 expression was 68.2% in ADC (located in the nucleus) and 29.6% in SCC (cytoplasm staining). The presence of TTF-1 was associated with better OS in SCC and ADC ( P =0.000 and P =0.003, respectively). In SCC, an increased level of TTF-1 was associated with a longer disease-free survival (DFS). Positive TTF-1 expression was an independent favorable prognostic factor in SCC ( P =0.020, HR: 2.789, 95%CI: 1.172-6.637) and ADC ( P =0.025, HR: 1.680, 95%CI: 1.069-2.641). TTF-1 was largely located in the nucleus of ADC, while it always accumulated in the cytoplasm of SCC. The higher level of TTF-1 in the different subcellular locations of ADC and SCC was an independent, favorable prognostic factor, respectively. Increased TTF-1 in the cytoplasm of SCC was associated with a longer OS and DFS.


Dual 3'Seq using deepSuperSAGE uncovers transcriptomes of interacting Salmonella enterica Typhimurium and human host cells.
The interaction of eukaryotic host and prokaryotic pathogen cells is linked to specific changes in the cellular proteome, and consequently to infection-related gene expression patterns of the involved cells. To simultaneously assess the transcriptomes of both organisms during their interaction we developed dual 3'Seq, a tag-based sequencing protocol that allows for exact quantification of differentially expressed transcripts in interacting pro- and eukaryotic cells without prior fixation or physical disruption of the interaction. Human epithelial cells were infected with Salmonella enterica Typhimurium as a model system for invasion of the intestinal epithelium, and the transcriptional response of the infected host cells together with the differential expression of invading and intracellular pathogen cells was determined by dual 3'Seq coupled with the next-generation sequencing-based transcriptome profiling technique deepSuperSAGE (deep Serial Analysis of Gene Expression). Annotation to reference transcriptomes comprising the operon structure of the employed S. enterica Typhimurium strain allowed for in silico separation of the interacting cells including quantification of polycistronic RNAs. Eighty-nine percent of the known loci are found to be transcribed in prokaryotic cells prior or subsequent to infection of the host, while 75% of all protein-coding loci are represented in the polyadenylated transcriptomes of human host cells. Dual 3'Seq was alternatively coupled to MACE (Massive Analysis of cDNA ends) to assess the advantages and drawbacks of a library preparation procedure that allows for sequencing of longer fragments. Additionally, the identified expression patterns of both organisms were validated by qRT-PCR using three independent biological replicates, which confirmed that RELB along with NFKB1 and NFKB2 are involved in the initial immune response of epithelial cells after infection with S. enterica Typhimurium.


Exosomal MiR-381 from M2-polarized macrophages attenuates urethral fibroblasts activation through YAP/GLS1-regulated glutaminolysis.
Post-traumatic urethral stricture is a clinical challenge for both patients and clinicians. Targeting glutamine metabolism to suppress excessive activation of urethral fibroblasts (UFBs) is assumed to be a potent and attractive strategy for preventing urethral scarring and stricture. In cellular experiments, we explored whether glutaminolysis meets the bioenergetic and biosynthetic demands of quiescent UFBs converted into myofibroblasts. At the same time, we examined the specific effects of M2-polarized macrophages on glutaminolysis and activation of UFBs, as well as the mechanism of intercellular signaling. In addition, findings were further verified in vivo in New Zealand rabbits. It revealed that glutamine deprivation or knockdown of glutaminase 1 (GLS1) significantly inhibited UFB activation, proliferation, biosynthesis, and energy metabolism; however, these effects were rescued by cell-permeable dimethyl α-ketoglutarate. Moreover, we found that exosomal miR-381 derived from M2-polarized macrophages could be ingested by UFBs and inhibited GLS1-dependent glutaminolysis, thereby preventing excessive activation of UFBs. Mechanistically, miR-381 directly targets the 3'UTR of Yes-associated protein (YAP) mRNA to reduce its stability at the transcriptional level, ultimately downregulating expression of YAP, and GLS1. In vivo experiments revealed that treatment with either verteporfin or exosomes derived from M2-polarized macrophages significantly reduced urethral stricture in New Zealand rabbits after urethral trauma. Collectively, this study demonstrates that exosomal miR-381 from M2-polarized macrophages reduces myofibroblast formation of UFBs and urethral scarring and stricture by inhibiting YAP/GLS1-dependent glutaminolysis.


Patient ocular conditions and clinical outcomes using a PROSE scleral device.
To determine the type and distribution of ocular conditions cared for in a clinic dedicated to scleral devices and to report the clinical outcomes afforded by this approach. Fifty-one charts of patients fitted unilaterally or bilaterally with a scleral device (Prosthetic Replacement of the Ocular Surface Ecosystem - PROSE) in a two year period were retrospectively reviewed. Patient demographics, ocular diagnoses, associated systemic conditions, best corrected visual acuity (BCVA) before and after fitting, Visual Function Questionnaire score (VFQ-25), and ocular surface disease index (OSDI) score were collected. All 51 patients were successfully wearing the PROSE device for a period of anywhere from weeks to years. The most common reasons for fitting were to relieve symptoms of moderate to severe dry eye syndrome ("DES", n=25), management of refractive problems ("refractive", n=23) with keratoconus being the most common (n=14), and to manage other anomalies ("other", n=3). Best corrected visual acuity (logMAR) improved with the wearing of the PROSE device for both the DES (17 letters) and the refractive group (10 letters), but not the "other" group. No serious complications were recorded for any of the patients. The PROSE device is a useful option not only for the management of ocular surface disease and optical imperfections, but also for other ophthalmic conditions. Moderate to severe dry eye was the most common anomaly managed, followed by eyes with irregular corneal astigmatism. DES and refractive patients experienced improvement in visual acuity with wearing of the PROSE device.


Repeatability of pachymetric mapping using fourier domain optical coherence tomography in corneas with opacities.
To evaluate the repeatability of Fourier domain optical coherence tomography (OCT) pachymetric mapping in patients with corneal opacities and to assess the reliability of Fourier domain OCT with 830 nm wavelength as a pachymetric measurement tool in opaque corneas. A Fourier domain OCT system was used to map the corneal thickness of patients with corneal scars or dystrophy. A retrospective study of a consecutive series was conducted. The repeatability was measured using pooled standard deviation of repeated measurements. A slit-scanning tomography device provided pachymetric mapping for comparison. Seventeen eyes of 12 patients with corneal scars (7 trauma and 3 post infection) or dystrophy (2 Reis-Bucklers and 5 granular dystrophy) were included. The posterior corneal boundary was detectable in all cases. The average corneal thickness measured by OCT was 536 ± 89 μm in central 2 mm area, 553 ± 76 μm in pericentral 2- to 5-mm area, and 508 ± 93 μm for the minimum corneal thickness. The slit-scanning tomography central corneal thickness, 433 ± 111 μm, was significantly lower than OCT readings (mean difference -91.1 ± 33.3 μm, P = 0.002). Repeatability of the OCT measurements was 2.1 μm centrally and 1.2 μm pericentrally. Pachymetric mapping with Fourier domain OCT was highly repeatable. Fourier domain OCT is a reliable pachymetric tool in opaque corneas. In comparison, corneal thickness measured by the slit-scanning tomography is significantly thinner than those measured by the Fourier domain OCT in the presence of corneal opacities.


Incidence of Leber hereditary optic neuropathy in 2019 in Japan: a second nationwide questionnaire survey.
Leber hereditary optic neuropathy (LHON) is an acute or subacute optic neuropathy that mainly affects young males. The first nationwide epidemiological survey of LHON was conducted in 2014 in Japan, and LHON was officially designated as a rare intractable disease by the Japanese government in 2015. We conducted a second survey of the annual incidence of LHON in 2019, and estimated the total number of patients with LHON in Japan. A questionnaire was sent to 997 facilities accredited by the Japanese Ophthalmological Society and/or affiliated with the councilors of the Japanese Neuro-Ophthalmology Society. Responses were received from 791 facilities, with a response rate of 79%. Fifty-five newly diagnosed cases (49 males and 6 females) of LHON were reported from 35 institutions in 2019, with a median age of 28.5 for males and 49.5 years for females. The total number of newly diagnosed cases was calculated as 69 (62 were males and 7 were females, 95% confidence interval 55-83), and the total number of patients was estimated to be 2491 (95% confidence interval: 1996-2986), suggesting a prevalence of LHON in Japan of 1:50,000. The incidence of LHON in 2019 was lower than the estimate in 2014, whereas its prevalence may be similar to that reported in other countries. The accurate estimation of the incidence and prevalence of patients with LHON requires prospective registration.


Displacement bone marrow transplantation for some inborn errors.
The initial simple bone marrow transplants for genetic immunodeficiency diseases could hardly be rejected by the host, but required matched sibling donors, only available for about 1 in 5 patients. Improved inductions enabled alternative donors from the family or unrelated volunteers to be used. Measurement of the extent of engraftment by donor cell markers or their normal enzymes showed the need for displacement, which aims to obtain 100% donor-type marrow so that the future immune responses of the recipient become those of the donor and tolerant to donor cells or their products. Immunoprophylaxis can prevent residual host immune cells from surviving to impair the graft. The concept of DBMT with immunoprophylaxis has evolved either to replace abnormal host cells or to confer a component transferable from donor cells to deficient host tissues. Within 10 years over 40 previously fatal genetic diseases have been satisfactorily corrected and seven partially corrected, but for five there has been inadequate delivery of component to genetically defective tissues such as heart, cartilage and brain. The principles can be applied to some 40 other genetic diseases for which no suitable alternative treatments yet exist.


Alternative techniques in cochlear implant surgery: Subtotal petrosectomy.
To report and analyze three cases of subtotal petrosectomy (SP) in cochlear implant surgery at our institution, and establish the indications, surgical technique and complications encountered. A retrospective descriptive study is proposed, analyzing a series of three clinical cases of subtotal petrosectomy as surgical technique for cochlear implant surgery at San Ignacio University Hospital (Bogotá, Colombia) from year 2004 to 2019. A total of three cases of subtotal petrosectomy as surgical technique in cochlear implant candidates were analyzed. The indications were the presence of a wide mastoid cavity after canal wall down mastoidectomy, extrusion of the electrode into the external auditory canal with a wide mastoid cavity and erosion of the posterior wall of the ear canal, and the presence of cholesteatoma in a cophotic ear with previous surgery. The ear canal was defunctionalized in all three cases; in two of them with obliteration of the Eustachian tube and in none of the cases the mastoid was obliterated. There was a single complication associated with the procedure corresponding to a small retention cholesteatoma in the skin of the obliterated duct sac, that didn't required surgical intervention. Subtotal petrosectomy is a surgical alternative for cochlear implant surgery in patients with chronic ear pathology, wide cavities or cochlear implant extrusion, not associated to significant complications.


Microarray analysis identification of key pathways and interaction network of differential gene expressions during osteogenic differentiation.
Adult bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent stem cells that can differentiate into three lineages. They are suitable sources for cell-based therapy and regenerative medicine applications. This study aims to evaluate the hub genes and key pathways of differentially expressed genes (DEGs) related to osteogenesis by bioinformatics analysis in three different days. The DEGs were derived from the three different days compared with day 0. Gene expression profiles of GSE37558 were obtained from the Gene Expression Omnibus (GEO) database. A total of 4076 DEGs were acquired on days 8, 12, and 25. Gene ontology (GO) enrichment analysis showed that the non-canonical Wnt signaling pathway and lipopolysaccharide (LPS)-mediated signaling pathway were commonly upregulated DEGs for all 3 days. KEGG pathway analysis indicated that the PI3K-Akt and focal adhesion were also commonly upregulated DEGs for all 3 days. Ten hub genes were identified by CytoHubba on days 8, 12, and 25. Then, we focused on the association of these hub genes with the Wnt pathways that had been enriched from the protein-protein interaction (PPI) by the Cytoscape plugin MCODE. These findings suggested further insights into the roles of the PI3K/AKT and Wnt pathways and their association with osteogenesis. In addition, the stem cell microenvironment via growth factors, extracellular matrix (ECM), IGF1, IGF2, LPS, and Wnt most likely affect osteogenesis by PI3K/AKT.


High-Complexity cellular barcoding and clonal tracing reveals stochastic and deterministic parameters of radiation resistance.
It is elusive whether clonal selection of tumor cells in response to ionizing radiation (IR) is a deterministic or stochastic process. With high resolution clonal barcoding and tracking of over 400 000 HNSCC patient-derived tumor cells the clonal dynamics of tumor cells in response to IR was analyzed. Fractionated IR induced a strong selective pressure for clonal reduction which significantly exceeded uniform clonal survival probabilities indicative for a strong clone-to-clone difference within tumor cell lines. IR induced clonal reduction affected the majority of tumor cells ranging between 96% and 75% and correlated to the degree of radiation sensitivity. Survival to IR is driven by a deterministic clonal selection of a smaller population which commonly survives radiation, while increased clonogenic capacity is a result of clonal competition of cells which have been selected stochastically. A 2-fold increase in radiation resistance results in a 4-fold (P < .05) higher deterministic clonal selection showing that the ratio of these parameters is amenable to radiation sensitivity which correlates to prognostic biomarkers of HNSCC. Evidence for the existence of a rare subpopulation with an intrinsically radiation resistant phenotype commonly surviving IR was found at a frequency of 0.6% to 3.3% (P < .001, FDR 3%). With cellular barcoding we introduce a novel functional heterogeneity associated qualitative readout for tracking dynamics of clonogenic survival in response to radiation. This enables the quantification of intrinsically radiation resistant tumor cells from patient samples and reveals the contribution of stochastic and deterministic clonal selection processes in response to IR.


Implications of phenothiazine side effects: a study of antiparkinsonian agents in an older population.
A study of anti-Parkinsonism agents in prolonged phenothiazine therapy seeks to clarify some anecdotal misconceptions. Antiparkinsonian agents do not appear to affect the therapeutic efficiency of phenothiazines, nor does using them on a "demand" basis increase the problems of control of parkinsonian side effects. Older patients, however, appear to require the extended usage of antiparkinsonian agents rather more than some studies would suggest. Sustained release antiparkinsonian agents may yet further simplify the management of schizophrenia.


Expression of Genes for Methylxanthine Pathway-Associated Enzymes Accompanied by Sex Steroid Receptor Status Impacts Breast Carcinoma Progression.
Consumption of methylxanthine alkaloids appears to induce activities by antagonizing adenosine receptors, implicated in breast cancer behavior in vitro. Our goal was to evaluate expression of genes for methylxanthine receptors and metabolizing enzymes to assess risk of breast carcinoma recurrence. Clinical outcomes, estrogen/progestin receptor results, and gene expression assays guided selection. RNA was isolated from laser capture microdissection-procured carcinoma cells for microarray using established protocols. Gene expression levels of eight methylxanthine receptors, eight metabolizing enzymes, and various phosphodiesterases were retrieved from microarray results. Univariable Cox regressions and Kaplan-Meier plots were determined for each gene with R software. Individually, lower expressions of PDE4A, CYP2A6, or CYP2E were related to decreased progression-free survival (PFS) and overall survival (OS). PDE1A over-expression predicted decreased PFS and OS. ADORA2B and RYR1 over-expressions predicted diminished OS. ER+ cancers exhibited lower ADORA1, ADORA2B, and RYR1 and elevated PDE4A, CYP2A6, and CYP2E expressions. Of PR+ carcinomas, diminished ADORA2B and RYR1 and elevated expressions of ADORA3, PDE4A, CYP2C8, and CYP2E were noted. Least absolute shrinkage and selection operator (LASSO) revealed that CYP2E, PDE1A, and PDE4A expressions collectively predicted PFS whereas ADORA1, CYP2E, PDE1A, PDE1B, and PDE4A expressions jointly predicted OS. Models were clinically significant when validated externally. LASSO also derived a six-gene model and five-gene model that predicted PFS of ER- or PR- carcinomas, respectively. Similarly, five-gene and four-gene models predicted OS in ER- or PR- carcinomas, respectively. Collectively, expression of genes involved in methylxanthine action and metabolism in single-cell types predicted clinical outcomes of breast carcinoma indicating promise for developing diagnostics and design of new therapeutics.


Cytogenetic effects of influenza virus infection on male germ cells of mice.
Swiss albino male mice were administered two doses (1 and 2 HA units) of influenza A2 Hong Kong/68 virus IP. The incidence of chromosomal anomalies in spermatocytes was analysed at various times post infection and was found to be significantly higher than in controls, indicating that the influenza virus had induced these anomalies.


Open-sandwich immunoassay for sensitive and broad-range detection of a shellfish toxin gonyautoxin.
At present, the analytical method for paralytic shellfish poisoning (PSP) toxins in shellfish is the mouse bioassay (MBA), which is an official method of the Association of Analytical Communities (AOAC [8]). However, the low sensitivity and concerns over the number of live animals required for testing have been cited as the major reason for seeking its replacement. In this report, we employed an open-sandwich immunoassay (OS-IA) to detect gonyautoxin (GTX2/3), a kind of PSP toxins. OS-IA, which utilizes the antigen-induced enhancement of antibody VH/VL interaction, can measure a small molecule antigen in a noncompetitive format. Hence it has a wider working range and shorter measurement time. We isolated anti-GTX2/3 antibody gene from a hybridoma GT-13A by screening a Fab-displaying phage library. Then the vectors for OS-IA were constructed, and examined for antigen concentration-dependency of the VH/VL interaction by OS-ELISA. As a result, in each case, signal intensity increases notably in a wide concentration range (0.1 to >1000 ng mL(-1)) of free GTX2/3, which was enough to cover its regulation value (80 μg 100 g(-1)) in many countries. So OS-IA will be widely applicable to detect PSP toxins in shellfish meats and in drinking water.


Different behavior of erythrovirus B19 and torquetenovirus in response to a single step of albumin purification.
The safety of human serum albumin (HSA) is of special interest with respect to virus transmission because of the wide use of this blood product as a therapeutic agent and also, added to other products, as an excipient or a stabilizer. Conflicting data are reported concerning HSA contamination by small, naked viruses such as the erythrovirus B19 (B19V) and the anellovirus torquetenovirus (TTV). This study has been performed to assess the effect of the HSA purification procedures on the viral contamination. Known concentrations of B19V and TTV virus were spiked in raw Fraction V, the starting material from fractionated human plasma for HSA purification, which was subsequently submitted to the depth filtration procedure. After spiking, B19V and TTV genome copies were determined by real-time quantitative polymerase chain reaction assays in the mixture at the end of Fraction V dissolution, to determine the virus concentration achieved, in the HSA solution after the filtration step, in the filtered postwashing fluid, and in the supernatant of resuspended Celite. B19V was completely adsorbed by the Celite used as a filter aid in the depth filtration process and was thus undetectable in the resulting HSA-containing fraction. In contrast, in 2 out of 3 experiments, TTV was detected in all samples. The different behavior of the two viruses might be a reflection of their different surface charge.


Drug resistant tuberculosis among elderly: Challenges.
The article deals with challenges faced by the geriatric populations while on MDR treatment. Risk factors like tobacco use, low socio-economic status, previous disease, longer delays in seeking treatment and reduced mobility are some of the challenges while initiating MDR treatment. Other issues like drug-related adverse events and increased co-morbidity pose a major challenge while treating patients. Susceptibility among the geriatric age group includes various anatomical and physiological changes including nutritional deficiencies and co morbidities.


Transfer of verocytotoxigenic Escherichia coli O157, O26, O111, O103 and O145 from fleece to carcass during sheep slaughter in an Irish export abattoir.
The purpose of this study was to investigate carriage and transfer of verocytotoxigenic Escherichia coli (VTEC) O157, O26, O111, O103 and O145 from fleece to dressed carcasses of 500 sheep, and to establish the virulence potential of recovered VTEC. Individual sheep were tracked and sampled (10 g fleece, full carcass swab) through the slaughter process. Samples were examined for the presence of verotoxin (vt1 and vt2) genes using a duplex real-time PCR assay and positive samples were further screened for the presence of the above five serogroups by real-time PCR. VTEC cells were recovered from PCR positive samples by serogroup specific immunomagnetic separation and confirmed by serogroup specific latex agglutination and PCR. Isolates were subject to a virulence screen (vt1, vt2, eaeA and hlyA) by PCR and isolates carrying vt genes were examined by Pulsed-Field Gel Electrophoresis (PFGE). VTEC O26 was recovered from 5/500 (1.0%) fleece and 2/500 (0.4%) carcass samples. VTEC O157 was isolated from 4/500 (0.8%) fleece samples and 3/500 (0.6%) carcass samples. E. coli O103 was recovered from 84/500 (16.8%) fleece and 68/500 (13.6%) carcasses, but only one E. coli O103 isolate (0.2%) carried vt genes. E. coli O145 was recovered from one fleece sample, but did not carry vt genes. E. coli O111 was not detected in any samples. For the four serogroups recovered, the direct transfer from fleece to carcass was not observed with PFGE showing that VTEC O26 isolates from a matched fleece/carcass "pair" were not identical. This study shows that while VTEC O157 are being carried by sheep presented for slaughter in Ireland, other potentially clinically significant verotoxin producing strains (particularly VTEC O26) are emerging.


A clinical study of a fibrinogen-based collagen fleece for dural repair in neurosurgery.
There is frequently a need for dural grafts to cover defects resulting from retraction, shrinkage, or excision following neurosurgical procedures. Many substances have been tried as possible dural substitution, and different tissues and materials have been evaluated for use in dural repair. The authors performed a retrospective review of 288 consecutive neurosurgical procedures using a fibrinogen based collagen fleece (TachoComb), a resorbable mesh of collagen from horse tendons, coated with human fibrinogen, bovine thrombin, bovine aprotinin and riboflavin (for marking the coated side), for dural substitution. The fibrinogen and thrombin imitate the last step of the coagulation cascade. On contact with bleeding wounds or other body fluids the coagulation factors dissolve and a link is formed between the collagen carrier and the wound surface. Thrombin converts fibrinogen into fibrin by splitting off peptides. Aprotinin prevents premature lysis of the fibrin clot by plasmin. Neither superficial or deep wound infections nor aseptic meningitis were noted. We found good fibrous incorporation of TachoComb into the surrounding normal dura. Postoperative cerebrospinal-fluid (CSF) leaks developed in only five cases, who had to be re-operated, upon as well as one patient with a rebleeding. In another four cases, there was notable subcutaneous cerebrospinal-fluid accumulation without CSF-leak. They required a lumbar cerebrospinal-fluid drainage. We conclude that TachoComb is a valuable alternative to the patients fibrous tissues for dural repair in cases in which autogenous tissues are either unavailable or insufficient for proper reconstruction.


Sensing of sound pressure gradients by C. elegans drives phonotaxis behavior.
Despite lacking ears, the nematode C. elegans senses airborne sound and engages in phonotaxis behavior, enabling it to locate and avoid sound sources.1 How worms sense sound, however, is not well understood. Here, we report an interesting observation that worms respond only to sounds emitted by small but not large speakers, indicating that they preferentially respond to localized sound sources. Notably, sounds emitted by small speakers form a sharp sound pressure gradient across the worm body, while sounds from large speakers do not, suggesting that worms sense sound pressure gradients rather than absolute sound pressure. Analysis of phonotaxis behavior, sound-evoked skin vibration, and sound-sensitive neuron activities further support this model. We suggest that the ability to sense sound pressure gradients provides a potential mechanism for worms to distinguish sounds generated by their predators, which are typically small animals, from those produced by large animals or background noise. As vertebrate cochlea and some insect ears can also detect sound pressure gradients, our results reveal that sensing of sound pressure gradients may represent a common mechanism in auditory sensation across animal phyla. VIDEO ABSTRACT.


[Chemical Constituents from Sphagneticola trilobata].
To study the chemical constituents of the whole plant of Sphagneticola trilobata. The compounds were isolated and purified by column chromatography and their structures were determined by spectroscopic techniques. Three lignans, two indolics and two phenolic glycosides were isolated from the whole plant of Sphagneticola trilobata and identified as syringaresinol-4-O-β-D-glucopyranoside(1), pinoresinol-4-sulfate(2), pinoresinol-4-O-β-D-glucopyranoside(3), 1H-indole-3-carboxylic acid (4), 1H-indole-3-carbaldehyde(5), 2,6-dimethoxy-4-hydroxyphenol-1-O-β-D-glucopyranoside (6), and 3,5-dimethoxy-4-hydroxyphenol-1-O-β-D-glucopyranoside (7). Compounds 1 - 7 are isolated from the genus Wedelia for the first time. Compound 4 demonstrates significant inhibitory activity against α-glucosidase.


Pulmonary, liver and cerebral paragonimiasis: An unusual clinical case in Colombia.
Mainly affecting poorer populations in remote (sub)tropical areas, paragonimiasis is considered one of the world's most neglected tropical diseases by the World Health Organization. It is usually acquired by consuming undercooked freshwater crustaceans and primarily affects the lungs, but may ectopically migrate to other organs to produce a multisystemic clinical presentation. This study details what appears to be the first documented case in South America and particularly in Colombia of cerebral paragonimiasis and infection by the crab Moreirocarcinus emarginatus. After consuming this crab, a 32-year-old Colombian male developed dyspnea and headache. A chest X-ray revealed tension pneumothorax (TPT) and a thoracoscopic lung biopsy bronchiolitis, eosinophilia, and granulomatous reaction due to Paragonimus parasites. Brain tomography and MRI also showed a configuration typical of Paragonimus parasites, namely a predominantly cystic area near the left cuneus with diffusion restriction, a hemosiderin halo, and linear enhancement characteristic of the tunnel sign, indicating an adult worm moving in the brain. Cerebral paragonimiasis can occur in the Colombian population and Moreirocarcinus emarginatus can be its cause. In rural areas, education about raw crab consumption would be beneficial, and physicians addressing lung-related complaints should bear paragonimiasis in mind and ask about raw crab consumption.


Phenolamide and flavonoid glycoside profiles of 20 types of monofloral bee pollen.
This study aimed at investigating phenolamides and flavonoid glycosides in 20 types of monofloral bee pollen. The plant origins of pollen samples were determined by DNA barcoding, with the purities to over 70 %. The 31 phenolamides and their 33 cis/trans isomers, and 25 flavonoid glycosides were identified; moreover, 19 phenolamides and 14 flavonoid glycosides as new-found compounds in bee pollen. All phenolics and flavonoids are present in the amidation or glycosylation form. The MS/MS cleavage modes of phenolamides and flavonoid glycosides were summarized. Isorhamnetin-3-O-gentiobioside presented the highest levels 23.61 mg/g in apricot pollen. Phenolamides in 11 types of pollen constituted over 1 % of the total weight, especially 3.9 % in rose and 2.8 % in pear pollen. Tri-p-coumaroyl spermidine and di-p-coumaroyl-caffeoyl spermidine respectively accounted for over 2.6 % of the total weight in pear and rose pollen. The richness in phenolamides and flavonoid glycosides can offer bee pollen more bioactivities as functional foods.


In vitro studies in microsomes from rat and human liver, kidney, and intestine suggest that perfluorooctanoic acid is not a substrate for microsomal UDP-glucuronosyltransferases.
Perfluorooctanoic acid (PFOA) is a fluorinated fatty acid analogue used as a surfactant in the manufacture of fluoropolymers. Previous studies have indicated that PFOA was metabolically inert in mammals, but recent metabolism studies with related fluorochemicals suggested that PFOA might form a glucuronide conjugate. [(14)C(1)]-PFOA was incubated with male and female human and rat liver, kidney, and small intestine microsomes. Incubations were carried out in the presence of alamethicin and beta-saccharolactone to increase access of PFOA to the enzyme active site and to inhibit potential hydrolysis of PFOA-glucuronide by microsomal beta-glucuronidase, respectively. Although positive control experiments using p-nitrophenol demonstrated significant UDP-glucuronosyltransferase (UDPGT) activity in all of the tested microsomal preparations, no evidence for formation of a PFOA-glucuronide was obtained, either by high-sensitivity radiochromatography or by LC/MS. These data suggest that PFOA is not a substrate for human or rodent microsomal UDPGTs.


Ninety-five percent probability of paternity with HLA, ABO and haptoglobins.
Using HLA tissue typing as the basic genetic test along with ABO and haptoglobin typing, it is generally possible either to exclude a putative father conclusively or, in nonexclusion cases, to assign a greater than 95% probability of paternity. In 22 of 53 nonexclusion cases, the putative father had a probability of paternity above 99% based on these genetic tests.


Public health assessment for dioxins exposure from soil.
Dioxins are among the most toxic anthropogenic chemicals in the environment. Their toxicity has been extensively studied in both humans and animals. Dioxin-contaminated soil may result in dioxins occurring in a food chain. This is especially important for the general population. It has been estimated that about 98% of exposure to dioxins is through the oral route. In the 1980s, a concentration level of 1 ppb 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in soil was specified as "a level of concern," based on cancer effects. However, recent studies indicate that end points other than cancer are also of concern. A health risk analysis scenario based on health effects of TCDD other than cancer is discussed and compared with the projected intake from 1 ppb TCDD in soil.


Fos expression induced by olanzapine and risperidone in the central extended amygdala.
The extended amygdala has been proposed to play an essential role in cognitive and affective processes and in neuropsychiatric disorders. In the present study, we examined the induction of Fos-like nuclei in the central amygdaloid nucleus (CeA), sublenticular extended amygdala (SLEA), interstitial nucleus of the posterior limb of the anterior commissure (IPAC), and bed nucleus of the stria terminalis (BSTL) of rodents to improve the knowledge regarding the pharmacological profile, therapeutic efficacy, and side-effects of olanzapine, an atypical antipsychotic drug and risperidone, a mixed atypical/typical antipsychotic drug in the rat brain. In addition, we evaluated the induction of Fos-like-nuclei in areas connected with these structures such as prefrontal cortex (PFCx), and nucleus accumbens shell, and in other important areas including the lateral septum and caudate-putamen that are involved in the therapeutic efficacy or side-effects of antipsychotic drugs. Fos-like-immunoreactivity induced by olanzapine and risperidone was compared with that by the atypical antipsychotic clozapine and typical antipsychotic haloperidol. Regarding the extended amygdala, and similarly to clozapine, olanzapine (5-10 mg/kg) and, with a lower efficacy, risperidone (1-3 mg/kg), induced Fos-like-nuclei in CeA, IPAC, SLEA, and BSTL. Both these drugs increased the induction of Fos-like-nuclei in PFCx, nucleus accumbens shell, lateral septum, and caudate-putamen. On the contrary, the increase of Fos-like-nuclei in the extended amygdala by haloperidol was restricted to IPAC only. These findings, consistent with the important role of extended amygdala in neuropsychiatric disorders characterized by affective disturbances, showed that olanzapine and risperidone, contrary to haloperidol, preferentially activated Fos-expression in these brain areas.


The influence of vinpocetine alone or in combination with Epigallocatechin-3-gallate, Coenzyme COQ10, Vitamin E and Selenium as a potential neuroprotective combination against aluminium-induced Alzheimer's disease in Wistar Albino Rats.
Alzheimer's disease (AD) is one of such diseases that represent the most prominent cause of dementia in elderly people. To explore the possible neuroprotective effect as well as mechanism of action of Vinpocetine either alone or in combination with EGCG, CoQ10, or VE & Se in ameliorating aluminum chloride-induced AD in rats. Rats were received AlCl3 (70 mg/kg) intraperitoneal daily dose for 30 days along with EGCG (10 mg/kg, I.P), CoQ10 (200 mg/kg, P.O), VE (100 mg/kg, P.O) & Se (1 mg/kg, P.O) as well as Vinpocetine (20 mg/kg, P.O) either alone or in combination. Results revealed that the combination of Vinpocetine with EGCG showed the best neuroprotection. This protection in the brain was indicated by the significant decrease in Aβ and ACHE. The same pattern of results were shown in the levels of monoamines and BDNF. In addition, the combination of Vinpocetine with EGCG showed more pronounced anti-inflammatory (TNF-α, IL-1β) and antioxidant (MDA, SOD, TAC) effects in comparison to other combinations. These results were confirmed using histopathological examinations as well as DNA fragmentation assays. Vinpocetine with EGCG showed pronounced protection on neurons against AD induced by AlCl3 in rats.


Mutational analysis of the Escherichia coli K-12 TolA N-terminal region and characterization of its TolQ-interacting domain by genetic suppression.
The Tol-Pal proteins of Escherichia coli are involved in maintaining outer membrane integrity. They form two complexes in the cell envelope. Transmembrane domains of TolQ, TolR, and TolA interact in the cytoplasmic membrane, while TolB and Pal form a complex near the outer membrane. The N-terminal transmembrane domain of TolA anchors the protein to the cytoplasmic membrane and interacts with TolQ and TolR. Extensive mutagenesis of the N-terminal part of TolA was carried out to characterize the residues involved in such processes. Mutations affecting the function of TolA resulted in a lack or an alteration in TolA-TolQ or TolR-TolA interactions but did not affect the formation of TolQ-TolR complexes. Our results confirmed the importance of residues serine 18 and histidine 22, which are part of an SHLS motif highly conserved in the TolA and the related TonB proteins from different organisms. Genetic suppression experiments were performed to restore the functional activity of some tolA mutants. The suppressor mutations all affected the first transmembrane helix of TolQ. These results confirmed the essential role of the transmembrane domain of TolA in triggering interactions with TolQ and TolR.


The inhibitory effects of oxytocin on distal colonic contractile activity in rabbits are enhanced by ovarian steroids.
To study the effects of oxytocin on isolated rabbit distal colon and the regulation of ovarian steroids by its action. Muscle strips parallel to either the circular or the longitudinal fibres were excised and suspended in tissue chambers containing 5 mL Krebs solution (37 degrees C) and bubbled continuously with 95% O(2) and 5% CO(2). The effects of oxytocin on isometric spontaneous contractile responses were recorded. The effects of atosiban, tetrodotoxin, Mg(2+), progesterone and oestradiol on the oxytocin-induced response were also examined. Oxytocin (1, 10 and 100 nmol L(-1)) dose dependently decreased the area under the contraction curve of distal colonic smooth muscle strips. The oxytocin receptor antagonist atosiban blocked the oxytocin (10 nmol L(-1))-caused responses in a dose-dependent manner. Tetrodotoxin (10 micromol L(-1)) had no effect on the oxytocin-induced response. Mg(2+)-free Krebs solution attenuated the oxytocin-induced response, but oestradiol (0.1 micromol L(-1)) or progesterone (0.1 micromol L(-1)) increased the oxytocin-induced response. These results suggest that oxytocin inhibits the contractile motility of the distal colon, which is regulated by Mg(2+) and ovarian steroids.


Downbeat nystagmus with a pseudocycloid waveform: improvement with base-out prisms.
Downbeat nystagmus in primary position and oscillopsia resulted from nutritional deficiency during prolonged intravenous therapy of a patient with hyperemesis gravidarum. Wide bandwidth infrared oculography demonstrated a pseudocycloid nystagmus waveform with an increasing-velocity exponential slow phase. Because the oscillopsia decreased and the nystagmus was damped with convergence, visual acuity improved with the addition of base-out prisms to each spectacle lens.


Lipidization of human interferon-alpha: a new approach toward improving the delivery of protein drugs.
Human interferon-alpha (IFN-alpha), a 19.2 KD protein containing two disulfide bonds (cys1-cys98; cys29-cys138), was reduced and modified with a reversible lipidization agent. The product of the lipidization, PAL-IFN, was homogenous, with four palmitoyl moieties linked to the four Cys residues in the protein molecule via reversible disulfide linkages. The far-UV circular dichroism (CD) spectrum of PAL-IFN was virtually overlapped with that of IFN, indicating that the IFN structure was not altered by the modification. After iv injection in mice of 0.1 mg/kg of PAL-IFN, a low level of serum IFN activity was sustained for more than 8 h, while serum IFN activity was rapidly diminished to an undetectable level at 2 h post IFN injection at the same dose. Evidence suggested that IFN was slowly released from PAL-IFN into blood circulation upon reduction of the disulfide bonds in vivo. Furthermore, the liver-targeting effect of PAL-IFN was demonstrated by the observation that the level of 2'-5' oligoadenylate synthetase (OAS) expressed in the liver of mice treated with PAL-IFN was significantly higher than that with IFN. In conclusion, reversible lipidization can potentially achieve both a prolonged plasma half-life and an enhanced liver-targeting effect of IFN in antiviral therapy.


Imaging of posterior fossa epidermoid tumors.
Epidermoid tumors have similar radiologic characteristics with arachnoid cysts on routine imaging techniques. Since they warrant different therapeutic interventions, it is essential to differentiate the two pathologies and to assess operative results. The purpose of this study is to define the MR characteristics of epidermoid tumors in posterior fossa particularly on fluid-attenuated inversion recovery (FLAIR) and diffusion-weighted imaging (DWI). During a 10-year period, we operated 19 cases with epidermoid tumor of posterior fossa. The localization of the epidermoids was cerebellopontine angle in 16 patients, within the fourth ventricle in 2 patients and pineal region in 1 patient. Neuroimaging included CT in all patients, conventional MR in 14 patients and FLAIR and DWI in the last 6 patients. In the first 13 cases, diagnosis of epidermoid tumor was difficult on CT and conventional MR. Epidermoids appeared as heterogeneous hyperintense lesions on FLAIR and homogenous hyperintense lesions on DWI in the last 6 patients. While total resection of the tumor was achieved in 17 patients, subtotal resection was performed in 2 patients. Histopathological examinations of the specimens revealed epidermoid tumor in all cases. Advances in neuroradiology provide us advantages for planning the treatment modalities in epidermoid tumors of the posterior fossa. The difficulties in preoperative differential diagnosis and judgment for reoperation in the postoperative course have been decreased particularly via FLAIR and DWI.


Parrondo's paradox in quantum walks with time-dependent coin operators.
We show that a Parrondo paradox can emerge in two-state quantum walks without resorting to experimentally intricate high-dimensional coins. To achieve such goal we employ a time-dependent coin operator without breaking the translation spatial invariance of the system.


Lack of reactivation of tuberculosis in patients with psoriasis treated with secukinumab in a real-world setting of latent tuberculosis infection.
Some biologics for psoriasis, especially anti-tumor necrosis factor (TNF)-α therapies, may re-activate latent tuberculosis (TBC) infection with consequent morbidity and mortality. However, there is a low reported incidence of conversion to positive TBC status among patients with psoriasis treated with second-generation biologic therapies, particularly anti-interleukin (IL)-17 therapies such as secukinumab. To evaluate the safety profile of secukinumab in psoriasis patients with latent TBC infection. Real-life data were collected by retrospective chart review on patients with moderate-to-severe psoriasis who showed positivity for TBC screening at baseline and underwent secukinumab treatment for psoriasis at six Italian centers. Patients received secukinumab 300 mg at week 0/1/2/3/4, then every 4 weeks. Fifty-nine patients were enrolled; 30.5% also had psoriatic arthritis and other comorbidities were common. At baseline, the mean psoriasis duration was 14.5 years. Ten (17%) patients did not undergo prophylaxis before starting secukinumab. Conversely, isoniazid ± rifampicin or rifampicin alone prophylaxis was administered in 49/59 (83.1%) patients. After a mean treatment duration of 84 weeks, there were no cases of TBC reactivation and no unexpected safety signals. Secukinumab use over an extended period was safe in psoriasis patients with latent TBC, even in patients who did not receive chemoprophylaxis.