Late effects of early binocular visual deprivation on the function of Brodmann's area 7 of monkeys (Macaca arctoides).
It has been shown earlier that binocular visual deprivation during the early sensitive period of life reduces the representation of visual functions in the posterior parietal association cortex of monkeys, in Brodmann's area 7 (Exp. Brain Res., 42 (1981) 1-8). Moreover, the representation of somatic functions increases suggesting that competitive mechanisms between the inputs from different modalities function during the early sensitive period of life in area 7. The aim of the present study was to find out whether further reorganization of functions takes place in the posterior parietal association cortex if monkeys that have experienced binocular visual deprivation through the first year of their life, are allowed to recover from the deprivation for a longer period of time. Four monkeys were deprived of binocular vision after the birth by lid closure for 12 months. Transdural extracellular multiunit recordings were performed in Brodmann's area 7 at the end of the deprivation period after the opening of the eyes. A second set of recordings was conducted in area 7 after a recovery period of 12 months from the deprivation. The results of the recordings at the end of the deprivation period confirmed the already known deprivation effect: there was a reduction in visually responsive neurons, an increase in the representation of somatosensory and somatomotor functions as well as an increase in the amount of cell groups that were 'only spontaneously active'. The recordings performed after the recovery period showed that the representation of visual functions had remained low. However, the amount of 'only spontaneously active' neurons had decreased and the amount of cell groups responding to the monkey's own explorative movements of the hand had further increased. The results indicate that visual deprivation during the early critical period of life results in a profound and persistent reduction of visual functions in area 7. However, activity-dependent competition between inputs from different modalities continue, resulting in the domination of somatosensory and somatomotor functions over visual functions in area 7. The results also suggest that neurons which during the visual deprivation are left without active input from the visual system, gradually become integrated into other functionally active neuronal networks increasing the representation of somatic functions in this cortical area.


KRAS and BRAF mutation analysis in colorectal adenocarcinoma specimens with a low percentage of tumor cells.
The rapid development of molecular biology techniques allows for the introduction of real-time polymerase chain reaction (PCR) methods with a limit of mutation detection at 1% in a background of wild-type DNA. Analysis of KRAS mutations in codons 12, 13, and 61, together with analysis of BRAF mutations in codon 600, are predictive biomarkers for anti-epidermal growth factor receptor (EGFR) treatment in colorectal cancer. Our aim was to compare PCR methods for KRAS mutations and BRAF mutation analysis using DNA isolated from tissue samples previously evaluated for presence of tumor cells using a quantitative scale and the percentage of tumor cells (PTC) scale. We addressed the question of whether a low number of tumor cells can be qualified for somatic mutation testing. Our study showed that PTC as low as 10% was good enough to detect KRAS G12D, G13D, and Q61L mutations in formalin-fixed paraffin-embedded (FFPE) material. Furthermore, our results indicate that up to 20% of colorectal cancer may carry mutations in the KRAS codon 61 and BRAF codon 600, which suggests the value of these mutation analyses because patients carrying them are unlikely to respond to cetuximab or panitumumab. A low level of KRAS somatic mutation detection has not been studied in depth in the context of clinical outcomes in patients; therefore, we compared new PCR methods, (KRBR-RT 50 Entrogen; ViennaLab StripAssay) and re-evaluated KRAS and BRAF status in patients with relapse after targeted therapy. The importance of molecular results was confirmed by clinical observation of a patient with relapse who had qualified for targeted therapy with KRAS WT status (but was diagnosed by less sensitive single-stranded conformation polymorphisms method). Interestingly, during anti-EGFR treatment, it came to the selection of cells with KRAS G12C mutation which were present from the beginning in the tumor but at a low level (detected by PCR methods) only and led consequently to the metastasis. Taking into consideration the limit of detection, labor time, and assay cost, the real-time PCR method seems to be very promising especially for FFPE material with the PTC below 15%.


Early versus delayed initiation of continuous negative pressure in infants with hyaline membrane disease.
Twenty-three infants with HMD of similar severity, who were less than 24 hours of age and who were breathing spontaneously, were divided by random numbers into early and delayed CNP groups. The infants who were treated with CNP before their PaO2 was less than 50 mm Hg while breathing 70% oxygen experienced a significantly greater increase in PaO2 in response to the initiation of CNP, required less time with O2 therapy, required no mechanical ventilation, and had fewer complications. Based on these results, it is suggested that CNP be initiated in infants with HMD, who are less than 24 hours of age and are breathing spontaneously, before the PaO2 becomes less than 50 mm Hg on 70% O2.


Sensitivities of radioimmunoassay and enzyme-linked immunosorbent assay for detection of hepatitis B surface antigen.
Radioimmunoassay (RIA) using Austria II (Abbott) was found to be more sensitive for the detection of HBsAg than the Hepanostika Micro-Elisa system (Organon Teknika), by a factor of 5- to 20-fold for subtype 'ad' (with one exception), and 2- to 5-fold for subtype 'ay'. The sensitivities of RIA and the enzyme-linked immunosorbent assay (Elisa) have also been evaluated in the screening of 178 selected serum samples contributed by 7 different laboratories. Of these, 149 were found to be positive for HBsAg by RIA, 142 were positive by Elisa and 36 were positive by CEP.


DNA methylation and histone modifications in breast cancer.
Breast cancer is caused by progressive genetic alterations and epigenetic changes that trigger tumor development. Among the epigenetic alterations descried in breast cancer, DNA promoter methylation has been extensively studied and observed in genes involved in several critical signaling pathways that initiate and promote breast tumorigenesis. In addition to DNA hypermethylation, global methylation levels have been observed to decrease with breast cancer progression. In close connection with DNA methylation changes, histone modifications, especially the acetylation and methylation of histone lysines, play an essential role in the nucleosomes remodeling and gene expression regulation in breast cancer.


Study of efficacy and safety of noncultured, extracted follicular outer root sheath cell suspension transplantation in the management of stable vitiligo.
Noncultured, extracted follicular outer root sheath suspension (NC-EHF-ORS-CS) is a recently introduced technique for the treatment of stable vitiligo. To study the clinical efficacy of this technique and to determine the viability and cell composition of the suspension. Twenty-five patients with stable vitiligo were included in this prospective study. Fifty follicles were extracted from occipital scalp and were incubated with trypsin-ethylenediaminetetraacetic acid to separate outer root sheath cells. The cell suspension was filtered and centrifuged to obtain a cell pellet, which was resuspended and applied to the dermabraded recipient area. Cell viability of the suspension was assessed using trypan blue staining, and markers of keratinocyte stem cells (CD200) and melanocytes (S100) were evaluated using flow cytometry and immunocytochemistry, respectively. At 6 months, the mean (±SD) repigmentation was 52 ± 25.1%, and >75% repigmentation was seen in 8/25 (32%) patients. Mean percentage cell viability of the suspension was 80 ± 17.2% with a mean concentration of CD200 + and S100 + cells being 7.91 ± 8.68% and 9.93 ± 1.22% (n = 3), respectively. Recipient site infection was seen in 4 of 25 (16%) patients and a color mismatch in 11 of 25 (44%) patients. NC-EHF-ORS-CS is a useful minimally invasive therapy for vitiligo.


Experimental and theoretical study of the vibrational spectra of 12-thiacrown-4 and 18-thiacrown-6, evaluation of the performance of the different anharmonic force fields compared to the scaled quantum mechanical force fields.
We report, to the best of our knowledge, for the first time the vibrational, IR and Raman, spectra of 12-thiacrown-4 (12t4) and 18-thiacrown-6 (18t6). To predict in what conformation 12t4 and 18t6 exist, for the vibrational analysis of both molecules and to assess the performance of the different computational methods for the accurate prediction of the vibrational frequencies of relatively large molecules, the computations were done using the harmonic and anharmonic force fields using the 6-31G* and 6-311G** basis sets. The computations were performed at the HF, B3LYP, CAM-B3LYP, BLYP, BP86, G96LYP, PBE1PBE, TPSSH and MP2 levels. Comparison was made between the calculated and experimental vibrational frequencies as indicated by the root-mean-square (rms) deviations, using either the unscaled and scaled harmonic vibrational frequencies and, unscaled, anharmonic vibrational frequencies. For the harmonic vibrational frequencies two scaling schemes were used. One uses one-scale-factor (1SF) scaling and the other uses 8SF scaling. In terms of the vibrational analysis of 12t4 and 18t6, the report confirms the solid state X-ray structure of D4 of 12t4 and C2 of 18t6. It is concluded that a lower rms deviation is obtained using 1SF scaled harmonic vibrational frequencies at even the HF/6-31G* level than using anharmonic vibrational frequencies at the MP2/6-311G** level. The CAM-B3LYP method showed some improvement over the traditional B3LYP method.


Studies in fetal wound healing: II. A fetal environment accelerates fibroblast migration in vitro.
We have used an in vitro model of wound healing using scratches made in a confluent monolayer of fibroblasts. The effects of fetal calf and postnatal calf serum on the migration of fibroblasts were compared. Differences between fetal and calf serum-incubated fibroblasts grown on coverslips were observed within 15 minutes of exposure. Cells in fetal serum began to change both shape and orientation and to move into the trough created by the scratch. The fibroblasts incubated in fetal calf serum completely filled in the trough within 16 hours while those incubated in calf serum did not do so even after 24 hours. We estimate that, at any point, there was a 50% lag time in the migration of the fibroblasts in the presence of postnatal calf serum. This difference in migration and filling was not a function of mitogenesis; the mitogenicity of the two sera were comparable. The results suggest that fibroblast migration in vitro is accelerated by the fetal serum. A similar mechanism may occur in vivo and may underlie the ability of the fetal wound to heal more rapidly.


Late results of triple valve replacement: a 14-year review.
Forty-eight patients have undergone simultaneous triple valve replacement at our institution over the past 14 years. This series was reviewed as a baseline for comparison with more recent tricuspid valve reparative procedures and to assess the natural history of advanced rheumatic heart disease with surgically corrected valvular lesions. Survival was 81% at one month and 32% at 14 years, not significantly different from results with double valve replacement during the same period, and not related to whether the tricuspid involvement was organic or functional. Forty-three percent of the late deaths were clearly valve related, a percentage which would undoubtedly be lowered with currently available prostheses. Functional class was determined in 13 of the 17 current survivors (mean follow-up, 10.3 years), and was improved over preoperative functional class in all but 3 patients. Three patients died of progressive congestive heart failure despite normally functioning prostheses; the reasons for these deaths are open to speculation.


Predictive value of qsofa in early mortality risk stratification in acute pulmonary embolism.
Pulmonary Embolism Severity Index (PESI) and simplified PESI (sPESI) methods are used in the evaluation of patients with suspected acute pulmonary thromboembolism (PTE). This study aimed to provide a rapid mortality risk stratification in patients with acute pulmonary thromboembolism (PTE) immediately after admission without relying on laboratory data by using quick sequential organ failure assessment (qSOFA), a three-parameter scoring system with proven efficiency used for swift prediction of organ dysfunction, and compare it with Pulmonary Embolism Severity Index (PESI) and simplified PESI (sPESI). This study included outpatients and inpatients diagnosed with acute PTE in our clinic and whose PESI, sPESI and qSOFA scores were calculated for early mortality risk classification. A total of 123 patients who were objectively diagnosed with PTE and followed up were prospectively observed. When their qSOFA scores were compared with the early mortality risk stratification in acute PTE, patients with a high qSOFA score were determined to be in the high-risk group in the early mortality risk stratification (p < 0.001). Overall, 69.2% of 26 patients with a high qSOFA risk (≥2) were found to be in the high-risk group in the early mortality risk binary stratification in acute PTE (p < 0.0001). The qSOFA score provides guidance to identify patients with acute PTE with potentially life-threatening hemodynamic decompensation or collapse in need of reperfusion therapy.


Integrated therapy decreases the mortality of patients with polymyositis and dermatomyositis: A Taiwan-wide population-based retrospective study.
The issue of whether integrated treatment with conventional medicine (CM) and herbal medicine (HM) can reduce mortality in patients with polymyositis/dermatomyositis (PM/DM) had not been addressed. In this study, we investigated the effect of integrated therapy on mortality in a retrospective PM/DM cohort in the Taiwan National Health Insurance Research Database (NHIRD). Patients with PM/DM were retrospectively enrolled from the PM/DM Registry of Catastrophic Illnesses cohort in the Taiwan NHIRD between 1997 and 2011. The patients were divided into an integrated medicine (IM) group that received CM and HM and a non-IM group that received CM alone. The Cox proportional hazards regression model and Kaplan-Meier method were used to evaluate the hazard ratio (HR) for mortality. Three hundred and eighty-five of 2595 patients with newly diagnosed PM/DM had received IM and 99 had received non-IM. The adjusted HR for mortality was lower in the IM group than in the non-IM group (0.42, 95% confidence interval 0.26-0.68, p < 0.001). The adjusted HR for mortality was also lower in the IM group that had received CM plus HM than in the group that received CM alone (0.48, 95% confidence interval 0.28-0.84, p < 0.05). The core pattern of HM prescriptions integrated with methylprednisolone, methotrexate, azathioprine, or cyclophosphamide to decrease mortality included "San-Qi" (Panax notoginseng), "Bai-Ji" (Bletilla striata), "Chen-Pi" (Citrus reticulata), "Hou-Po" (Magnolia officinalis), and "Dan-Shan" (Salvia miltiorrhiza). Integrated therapy has reduced mortality in patients with PM/DM in Taiwan. Further investigation of the clinical effects and pharmaceutical mechanism involved is needed.


Exclusive transition state stabilization in the supramolecular catalysis of Diels-Alder reaction by a uranyl salophen complex.
Whereas the parent uranyl salophen is catalytically inactive, its phenyl derivative effectively catalyses with turnover the reaction of benzoquinone with 1,3-cyclohexadiene, while showing no appreciable affinity towards reactants and product.


[Acute orthostatic intracranial hypotension].
The case of a young female with an acute idiopathic low cerebrospinal fluid pressure is presented. Clinical-imaging manifestations were typical and two consecutive blood patches resulted in a complete and durable resolution of the symptomatology. We present a review of the various investigation and therapeutic techniques.


The beta subunit of eukaryotic translation initiation factor 2 binds mRNA through the lysine repeats and a region comprising the C2-C2 motif.
Eukaryotic translation initiation factor 2 (eIF2) has been implicated in the selection of the AUG codon as the start site for eukaryotic translation initiation, since mutations in its three subunits in yeast that allow the recognition of a UUG codon by the anticodon of the initiator Met-tRNAMet have been identified. All such mutations in the beta subunit of eIF2 (eIF2beta) mapped to a region containing a putative zinc finger structure of the C2-C2 type, indicating that these sequences could be involved in RNA recognition. Another feature of eIF2beta that could mediate an interaction with RNA is located in the amino-terminal sequences and is composed of three repeats of seven lysine residues which are highly conserved in other species. We show here the ability of eIF2beta, purified from Escherichia coli as a fusion to glutathione S-transferase, to bind mRNA in vitro. Through a deletion analysis, mRNA binding was found to be dependent on the lysine repeats and a region encompassing the C2-C2 motif. Strong mRNA binding in vitro could be maintained by the presence of only one lysine or one arginine run but not one alanine run. We further show that only one run of lysine residues is sufficient for the in vivo function of eIF2beta, probably through charge interaction, since its replacement by arginines did not impair cell viability, whereas substitution for alanines resulted in inviable cells. mRNA binding, but not GTP-dependent initiator Met-tRNAMet binding, by the eIF2 complex was determined to be dependent on the presence of the lysine runs of the beta subunit.


Hepatitis B and C in the adult population of Bissau, Guinea-Bissau: a cross-sectional survey.
Hepatitis B virus (HBV) and hepatitis C virus (HCV) are prevalent in West Africa. To address the WHO 2030 goals of a 90% reduction in incidence and a 65% reduction in mortality for both infections, we assessed the prevalence of HBV and HCV from surveys in the general population. Participants in this cross-sectional survey were included from randomly selected houses in a demographic surveillance site in Bissau, Guinea-Bissau. Participants were interviewed and had a blood sample drawn for viral analyses (HBsAg, anti-HBs, anti-HBc, anti-HCV and HCV RNA). Risk factors of HBV and HCV infection were determined by binomial regression adjusted for sex and age. A total of 2715 participants were included in this study. The overall HBsAg prevalence was 18.7% (95% CI: 17.3-20.2%). HBsAg was associated with male sex (adjusted risk ratio (aRR): 1.64), and prevalence decreased with age >34 years. HBV exposure was found in 91.9% of participants. Although 72.6% of individuals without sexual debut had been exposed to HBV, ever engaging in a sexual relationship was associated with higher risk of HBV exposure (aRR 1.18). The anti-HCV prevalence was 0.5% (95% CI: 0.3-0.9%), and 78.6% of those had detectable HCV RNA. Risk factors for anti-HCV sero-positivity were age above 55 (aRR 10.60), a history of blood transfusion (aRR 5.07) and being in a polygamous marriage (aRR 3.52). In Guinea-Bissau initiatives to implement treatment and widespread testing are needed to reach the WHO 2030 goals.


[Naturo-economic aspects of ecologic agriculture strategy].
The most important problems related in agricultural production both in quality and value terms are their natural and market conditions, changes in food processing industry and in foreign trade. Opportunities for food production development and outlook for changing demand for food have been considered.


Multifocality of transitional cell carcinoma results from genetic instability of entire transitional epithelium.
Multifocality of transitional cell carcinoma (TCC) has been attributed to seeding of exfoliated tumor cells or to a general sensitivity of the entire urothelium to carcinogenic stimuli. By contrast, TCC has been shown to evolve as a consequence of genetic defects and chromosomal instability. We analyzed chromosomal patterns, total DNA content, and p53 and Ki67 expression in malignant and normal transitional cells to evaluate their relationship to the development of multifocal TCC. Included in the study were 47 patients, 16 women and 31 men, with a mean age of 70.04 years (range 37 to 83). Of 47 patients, 45 had TCC of the urinary bladder and 7 of those had synchronous ureteral involvement. Two patients had ureteral TCC and a history of TCC of the bladder. Using fluorescence in situ hybridization, numerical aberrations of chromosomes 7, 9, and 17 were detected in imprint specimens of histologically verified tumor and "normal" urothelium and were compared with static ploidy and p53 and Ki67 expression. Chromosome 7 was altered in 93.6%, chromosome 9 in 63.8% (including monosomy), and chromosome 17 in 87.2% of the 47 analyzed tumor and normal imprints. Differences between tumor and normal epithelium were observed in aberrational frequencies (number of cells showing chromosomal aberrations calculated on 200 cells counted, given in percentages). DNA content was aneuploid in all tumor specimens, but diploid in 20 (42.5%) of 47 normal specimens, according to lower aberration frequencies in these patients. p53 detection was positive in 82.9% of the tumor specimens and 76.6% of the normal specimens. Ki67 was positive in 87.2% of the tumor imprints and in 72.3% of the normal specimens. These data suggest a general genetic instability as a reason for multifocality in the entire transitional epithelium.


A SARS-CoV-2 spike ferritin nanoparticle vaccine protects hamsters against Alpha and Beta virus variant challenge.
The emergence of SARS-CoV-2 variants of concern (VOC) requires adequate coverage of vaccine protection. We evaluated whether a SARS-CoV-2 spike ferritin nanoparticle vaccine (SpFN), adjuvanted with the Army Liposomal Formulation QS21 (ALFQ), conferred protection against the Alpha (B.1.1.7), and Beta (B.1.351) VOCs in Syrian golden hamsters. SpFN-ALFQ was administered as either single or double-vaccination (0 and 4 week) regimens, using a high (10 μg) or low (0.2 μg) dose. Animals were intranasally challenged at week 11. Binding antibody responses were comparable between high- and low-dose groups. Neutralizing antibody titers were equivalent against WA1, B.1.1.7, and B.1.351 variants following two high dose vaccinations. Dose-dependent SpFN-ALFQ vaccination protected against SARS-CoV-2-induced disease and viral replication following intranasal B.1.1.7 or B.1.351 challenge, as evidenced by reduced weight loss, lung pathology, and lung and nasal turbinate viral burden. These data support the development of SpFN-ALFQ as a broadly protective, next-generation SARS-CoV-2 vaccine.


Bias and sample size in intensive care unit trials: Protocol for a meta-epidemiological study.
Systematic errors (bias) and random errors result in inflated and imprecise intervention effect estimates in randomised clinical trials (RCT) and meta-analyses. We aim to assess time trends in the Cochrane risk of bias domains and sample size in RCTs of intensive care unit (ICU) interventions. We will conduct a meta-epidemiologic study of RCTs included in Cochrane systematic reviews assessing any intervention used in adult patients admitted to the ICU. We will search the Cochrane Database of Systematic Reviews and include reviews published from March 2011 corresponding to the latest update in the Cochrane risk of bias tool. We will extract data on risk of bias judgments in the seven Cochrane bias domains and trial sample sizes and evaluate time trends using run charts. The primary outcome will be time trends in the annual proportion of trials with overall low risk of bias (low risk of bias in all bias domains). The secondary outcomes include time trends in the annual median trial sample size, and the annual proportion of trials with low-, unclear- and high risk of bias in each of the seven Cochrane bias domains. The outlined meta-epidemiologic study will assess time trends in risk of bias and sample sizes in RCTs assessing ICU interventions. This will inform researchers, healthcare personnel and policymakers on the general reliability of findings from RCTs of ICU interventions over time, and inform future RCT design and reporting.


NAD- and NADP-dependent 7alpha-hydroxysteroid dehydrogenases from bacteroides fragilis.
Twenty strains of Bacteroides fragilis were screened for hydroxysteroid oxidoreductase activity in cell-free preparations. Eighteen strains were shown to contain NAD-dependent 7alpha-hydroxysteroid dehydrogenase. Sixteen of the strains containing the NAD-dependent enzyme also contained NADP-depedent 7alpha-hydroxysteroid dehydrogenase, but invariably in lesser amounts. A strain particulary rich in both 7alpha-hydroxysteroid dehydrogenase activities was selected for further study. Measurement of activity as a function of pH revealed a fairly sharp optimal activity range of 9.5--10.0 for the NAD-dependent enzyme and a broad flat optimal range of 7.0--9.0 for the NADP-dependent enzyme. Michaelis constants for trihydroxy-bile acids for the NAD-dependent enzyme were in the range of 0.32--0.34 mM, whereas dihydroxy-bile acids gave a Km of 0.1 mM. Thin-layer chromatography studies on the oxidation product of 3alpha, 7alpha-dihydroxy-5beta-cholanoic acid (chenodeoxycholic acid) by the dehydrogenase revealed a band corresponding to that of synthetic 3alpha-hydroxy, 7-keto-5beta-cholanoic acid. Similarly the oxidation product of chenodeoxycholic acid by both 7alpha-hydroxysteroid dehydrogenase and commercially available 3alpha-hy-droxysteroid dehydrogenase revealed a band corresponding to that of synthetic 3,7-diketo-5beta-cholanoic acid. Neither of these two oxidation products could be distinguished from those by the Escherichia coli dehydrogenase oxidation previously reported. Disc-gel electrophoresis of a cell-free lyophilized preparation indicated one active band for NAD-dependent activity of mobility similar to that for the NADP-dependent E. coli enzyme. The NADP-dependent dehydrogenase was unstable and rapidly lost activity after polyacylamide disc-gel electrophoresis, ultracentrifugation, freezing on refrigeration at 4 degrees C. No 3 alpha- or 12alpha-oriented oxidoreductase activity was demonstrated in any of the strains examined.


The final player in the coenzyme A biosynthetic pathway.
The determination of the crystal structure of human phosphopantothenoylcysteine synthetase completes our knowledge of the enzyme structures involved in all steps of coenzyme A biosynthesis. This structure provides insight into the differences between bacterial and mammalian forms of the enzyme and may guide the structure-based development of novel antibacterial compounds.


Application of asymmetric flow field-flow fractionation (AsFlFFF) coupled to inductively coupled plasma mass spectrometry (ICPMS) to the quantitative characterization of natural colloids and synthetic nanoparticles.
A straightforward quantification method is presented for the application of asymmetric flow field-flow fractionation (AsFlFFF) combined with inductively coupled plasma mass spectrometry (ICPMS) to the characterization of colloid-borne metal ions and nanoparticles. Reproducibility of the size calibration and recovery of elements are examined. Channel flow fluctuations are observed notably after initiation of the fractionation procedure. Their impact on quantification is considered by using (103)Rh as internal reference. Intensity ratios measured for various elements and Rh are calculated for each data point. These ratios turned out to be independent of the metal concentration and total sample solution flow introduced into the nebulizer within a range of 0.4-1.2 mL min(-1). The method is applied to study the interaction of Eu, U(VI) and Th with a mixture of humic acid and clay colloids and to the characterization of synthetic nanoparticles, namely CdSe/ZnS-MAA (mercaptoacetic acid) core/shell-coated quantum dots (QDs). Information is given not only on inorganic element composition but also on the effective hydrodynamic size under relevant conditions. Detection limits (DLs) are estimated for Ca, Al, Fe, the lanthanide Ce and the natural actinides Th and U in colloid-containing groundwater. For standard crossflow nebulizer, estimated values are 7 x 10(3), 20, 3 x 10(2), 0.1, 0.1 and 7 x 10(-2) microg L(-1), respectively. DLs for Zn and Cd in QD characterization are 28 and 11 microg L(-1), respectively.


Adverse drug reactions of leukotriene receptor antagonists in children with asthma: a systematic review.
Asthma is the most common chronic condition of childhood. Leukotriene receptor antagonists (LTRAs) are included in international guidelines for children and young people (CYP), but there have been highly publicised concerns about potential adverse effects. The aim was to identify and understand the reported frequency of adverse drug reactions (ADRs) attributed to LTRAs in CYP with asthma. Embase, MEDLINE, PubMed and CINAHL were searched up to October 2020. Reference lists of eligible papers were manually screened. Eligible studies identified adverse events attributed to an LTRA in individuals aged between 0 and 18 years diagnosed with asthma. Four different tools were used to assess risk of bias or quality of data to accommodate the papers assessed. The search identified 427 papers after deduplication; 15 were included (7 case reports, 7 case-controlled or cohort studies and 1 randomised control trial (RCT)). 7012 patients were recorded, of which 6853 received an LTRA. 13 papers examined the ADRs attributed to montelukast, one to pranlukast and one to unspecified LTRAs. After language standardisation, 48 ADRs were found, 20 of which were psychiatric disorders. Across all studies, the most commonly reported ADRs were 'anxiety', 'sleep disorders' and 'mood disorders'. The frequency of ADRs could be calculated in seven of the eight studies. Applying standardised frequency terms to the prospective studies and RCT, there were 14 'common' and 'uncommon' ADRs. 'Common' ADRs included 'agitation/hyperactivity/irritability/nervousness', 'aggression' and 'headache'. The case reports showed a similar pattern, describing 46 different ADRs experienced by a total of eight patients. LTRAs have a wide range of suspected ADRs in CYP, predominantly gastrointestinal and neuropsychiatric disorders. Careful monitoring of CYP with asthma is required, both to assess and manage ADRs and to step treatment down when clinically stable. CRD42020209627.


Bioconcentration of polycyclic musks in fathead minnows caged in a wastewater effluent plume.
The synthetic polycyclic musks HHCB (Galaxolide®) and AHTN (Tonalide®) were monitored in fathead minnows (FHMs) caged for a month at various locations in the North Saskatchewan River (NSR), upstream and downstream of the Gold Bar wastewater treatment plant that serves the city of Edmonton, AB, Canada. In addition, the distribution of these musk compounds in the river was predicted using the fugacity-based Quantitative Water Air Sediment Interface (QWASI) model. In FHMs caged 0.15 km downstream of the wastewater outfall, mean concentrations of HHCB and AHTN were 7.4 and 0.4 μg g-1 wet weight, respectively. These are among the highest reported concentrations of these musk compounds in fish exposed to treated wastewater. The musk concentrations in FHMs were significantly lower further downstream of the outfall. High bioconcentration factors (BCFs) in FHMs that exceeded 104 higher than estimated concentrations in water indicated that there were low rates of biotransformation of the musks in the fish. In the FHMs caged at the site closest to the wastewater outfall, HHCB concentrations in FHMs were comparable to the body burdens that have been reported to moderate expression of vitellogenin in female rainbow trout, indicating that fish in the NSR downstream of the wastewater outfall may be at risk of anti-estrogenic effects. The QWASI model applied to six individual river sections of the NSR predicted that the largest fluxes of HHCB and AHTN would be for downstream transport in water, which explains why FHMs accumulated elevated concentrations of the musks at the furthest downstream site, 9.9 km from the wastewater discharge.


Influence of bone osteocalcin levels on bone loss induced by ovariectomy in rats.
To investigate the role of osteocalcin (OC) in bones, bone parameters in warfarin (WF)-treated rats after ovariectomy (OVX) were compared with those in intact rats. Rats were divided into an intact group and WF-treated group. Warfarin was orally given to rats for 16 weeks, and then OVX was performed and rats in the WF-treated groups continued receiving WF. Twelve weeks after OVX, bone properties were observed. The diaphysial bone OC level in the WF group was 10%-14% of the normal level at the preoperative point and 12 weeks after surgery. On comparison of the intact and WF groups before surgery, no significant differences were noted in bone mass parameters or mechanical properties, but 12 weeks after surgery, the diaphysial bone mineral content (BMC), bone area, and cortical thickness (Cth) were significantly higher in the WF-sham group than in the intact-sham group. Ovariectomy significantly decreased the diaphysial BMC, bone mineral density (BMD), Cth, and maximum load, and increased the endosteal perimeter in the WF group. In the intact group, no such OVX-induced changes were noted, and the metaphysial bone area and the endosteal and periosteal perimeters were increased by OVX. The CO(3)/PO(4) ratio in the femur measured by Fourier-transform infrared imaging using reflection preparations was higher in the WF-sham group than the intact-sham group, and higher in the intact-OVX group than the intact-sham group, but no significant difference was noted between the WF-sham and WF-OVX groups. It has been reported that CO(3)(-) is contained in new bone and decreases with mineral maturation. These data suggest that long-term reduction in bone OC levels may induce the formation of immature bone, which is easily resorbed with changes in bone metabolism such as OVX, and that OC may be one of the factors affecting bone turnover.


Diabetes-free survival among living kidney donors and non-donors with obesity: A longitudinal cohort study.
Approval of living kidney donors (LKD) with end-stage kidney disease (ESKD) risk factors, such as obesity, has increased. While lifetime ESKD development data are lacking, the study of intermediate outcomes such as diabetes is critical for LKD safety. Donation-attributable diabetes risk among persons with obesity remains unknown. The purpose of this study was to evaluate 10-year diabetes-free survival among LKDs and non-donors with obesity. This longitudinal cohort study identified adult, LKDs (1976-2020) from 42 US transplant centers and non-donors from the Coronary Artery Risk Development in Young Adults (1985-1986) and the Atherosclerosis Risk in Communities (1987-1989) studies with body mass index ≥30 kg/m2. LKDs were matched to non-donors on baseline characteristics (age, sex, race, body mass index, systolic and diastolic blood pressure) plus diabetes-specific risk factors (family history of diabetes, impaired fasting glucose, smoking history). Accelerated failure time models were utilized to evaluate 10-year diabetes-free survival. Among 3464 participants, 1119 (32%) were LKDs and 2345 (68%) were non-donors. After matching on baseline characteristics plus diabetes-specific risk factors, 4% (7/165) LKDs and 9% (15/165) non-donors developed diabetes (median follow-up time 8.5 (IQR: 5.6-10.0) and 9.1 (IQR: 5.9-10.0) years, respectively). While not significant, LKDs were estimated to live diabetes-free 2 times longer than non-donors (estimate 1.91; 95% CI: 0.79-4.64, p = 0.15). LKDs with obesity trended toward living longer diabetes-free than non-donors with obesity, suggesting within the decade following donation there was no increased diabetes risk among LKDs. Further work is needed to evaluate donation-attributable diabetes risk long-term.


Glycosylphosphatidylinositol (GPI) hydrolysis by transforming growth factor-beta1 (TGF-beta1) as a potential early step in the inhibition of epithelial cell proliferation.
Glycosylphosphatidylinositol (GPI) was previously identified in rabbit articular chondrocytes as being a precursor of inositolphosphate glycan (IPG), released upon (Transforming Growth Factor-beta) (TGF-beta) exposure, and capable of mimicking the proliferative effects of the growth factor. Here, using mink lung epithelial cells (CCL 64), which are known to be growth-inhibited by TGF-beta, we studied the potential role of GPI-derived molecules in the antiproliferative effect of TGF-beta1. We first identified an endogenous pool of GPI material and three different anionic forms of IPG in epithelial cells pre-labeled with [3H]glucosamine. Shortly (8 min) after TGF-beta1 addition, the cells responded by a rapid and transient hydrolysis of GPI, accompanied by the release of the most anionic form of IPG. This TGF-beta-released IPG, after partial purification, was shown to decrease the proliferation of CCL 64 cells. Moreover, anti-IPG antibodies reduced the effects of TGF-beta and blocked the effects of partially purified IPG. These data strongly suggest that GPI hydrolysis may be an early step of the TGF-beta signalling pathway involved in growth inhibition of epithelial cells.


The effect of oxygen toxicity and cigarette smoking on the binding of heterologous antibodies to alveolar basement membrane in the rat.
It has been suspected that pulmonary lesions of the Goodpasture type may be the result of circulating antibodies to alveolar basement membrane and that environmental factors such as cigarette smoke may influence antibody binding. Carefully designed experiments in this study have shown that exposure to cigarette smoke for 3 weeks or 100% oxygen for 65 h did not influence the binding of heterologous antibodies to alveolar basement membrane in Wistar rats, nor did these regimes increase any pathological changes associated with the antibody binding as detected by light microscopy. The results which are at variance with other reported findings are discussed with reference to reported animal models of antibody mediated pneumonitis and Goodpasture's syndrome in man.


The Emerging Role of the Single-Cell and Spatial Tumor Microenvironment in High-Grade Serous Ovarian Cancer.
The development of single-cell and spatial technologies has enabled a more detailed understanding of the tumor microenvironment and its role in therapy response and clinical outcome of high-grade serous ovarian cancer (HGSC). Interestingly, emerging evidence suggests that HGSCs with different genetic drivers harbor distinct tumor-immune microenvironments. Further, spatial cell-cell interactions have been shown to shape the CD8+ T-cell phenotypes and responses to immune checkpoint blockade therapies. The heterogeneous stroma consisting of cancer-associated fibroblast (CAF) subtypes, endothelia, and site-specific stromal types such as mesothelium modulates treatment responses via increasing stiffness and by producing ligands that promote drug resistance, angiogenesis, or immune escape. Chemotherapy itself shifts CAFs toward an inflammatory phenotype that associates with poor survival and immune-suppressive signaling. New emerging immunotherapies include combinational approaches and agents targeting, for example, the tumor-intrinsic endoplasmic reticulum pathway. A more detailed understanding of the spatial interplay of tumor, immune, and stromal cells in the tumor microenvironment is needed to develop more efficient immunotherapeutic strategies for HGSC.


Inequality amplifies the negative association between life expectancy and air pollution: A cross-national longitudinal study.
Ambient air pollution, in the form of fine particulate matter (PM2.5), poses serious population health risks. We estimate cross-national longitudinal models to test whether the negative relationship between life expectancy and PM2.5 concentration is larger in nations with higher levels of income inequality. The dependent variable is average life expectancy at birth, and the focal predictor variables include PM2.5 concentration, income inequality, and the two-way interaction between them. We also estimate the average marginal effects of PM2.5 concentration from low to high values of income inequality, and the predicted values of life expectancy from low to high values of PM2.5 concentration and income inequality. Results indicate that the negative relationship between life expectancy and PM2.5 concentration is larger in nations with higher levels of income inequality, and the reductions in predicted life expectancy are substantial when both PM2.5 concentration and income inequality are high. We suggest that the theoretical principles of Power, Proximity, and Physiology help explain our findings. This study underscores the importance in considering the multiplicative impacts of environmental conditions and socioeconomic factors in the modeling of population health.


Development of a heat-treated factor VIII/von Willebrand factor concentrate prepared from heparinized plasma.
A high yield, intermediate purity factor VIII concentrate derived from heparinized plasma has been developed which can be heat-treated at 60 degrees C, 68 degrees C or 80 degrees C/72 h to permit inactivation of viral contaminants which may be present. After cold reprecipitation of the heparinized cryoprecipitate (CRC), the resolubilized CRC precipitate was adjusted to 25-30 mg/ml protein and pH 6.35 +/- 0.1 and incubated for 1 h at 8 degrees C. After centrifugation to remove the precipitated fibrinogen and fibronectin, a factor VIII-rich supernatant can be recovered which contains greater than 500 units of VIII:C per liter of starting plasma (Method I product) at a purity of 1.5 U/mg protein. Adjusted to 50 mM glycine and pH 6.8, the product can be lyophilized and heat-treated at 60 degrees C/72 h without a significant loss of VIII:C activity. However, at 68 degrees C or 80 degrees C/72 h, temperatures now reported to be more effective in viral inactivation, the recoveries were reduced to 68 and 33% respectively. Significantly improved recoveries after heat-treatment (HT) at 68 degrees C or 80 degrees C/72 h were achieved if the 8 degrees C supernatant product was prepared by a modified procedure (Method II). This further reduces the fibrinogen content of the product while maintaining VIII:C yields greater than 500 U/l at a purity of 1.9 U/mg. When adjusted to 50 mM glycine and 1-2% (w/v) sucrose (pH 6.8), lyophilized and heat treated at 60 degrees C, 68 degrees C or 80 degrees C/72 h, the VIII:C recoveries of Method II product were 88-100%, 79-84% and 80-83% of pre-HT levels respectively. The yield of VIII:C was greater than 400 U/l at a purity of 1.6-1.4 U/mg at 1-2% (w/v) sucrose even after the severe heat-treatment at 80 degrees C. In addition, the von Willebrand factor multimers are similar in size and triplet pattern to those observed in routine cryoprecipitate preparations.


Cognitive training incorporating temporal information processing improves linguistic and non-linguistic functions in people with aphasia.
People with aphasia (PWA) often present deficits in non-linguistic cognitive functions, such as executive functions, working memory, and temporal information processing (TIP), which intensify the associated speech difficulties and hinder the rehabilitation process. Therefore, training targeting non-linguistic cognitive function deficiencies may be useful in the treatment of aphasia. The present study compared the effects of the novel Dr. Neuronowski® training method (experimental training), which particularly emphasizes TIP, with the linguistic training commonly applied in clinical practice (control training). Thirty four PWA underwent linguistic and non-linguistic assessments before and after the training as well as a follow-up assessment. Patients were randomly assigned to either experimental (n = 18) or control groups (n = 16). The experimental training improved both non-linguistic functions (TIP and verbal short-term and working memory) and linguistic functions: phoneme discrimination, sentence comprehension, grammar comprehension, verbal fluency, and naming. In contrast, the control training improved only grammar comprehension and naming. The follow-up assessment confirmed the stability of the effects of both trainings over time. Thus, in PWA, Dr. Neuronowski® training appears to have broader benefits for linguistic and non-linguistic functions than does linguistic training. This provides evidence that Dr. Neuronowski® may be considered a novel tool with potential clinical applications.


Study of pharmaceuticals in surface and wastewater from Cuernavaca, Morelos, Mexico: Occurrence and environmental risk assessment.
The present work describes the first known study to date on the occurrence of pharmaceuticals in surface water and wastewater of Cuernavaca, the capital of the state of Morelos (México). Selected pharmaceuticals (a total of 35) were extracted from the collected water samples with a generic solid phase extraction (SPE) protocol and determined in the sample extracts by means of high-performance liquid chromatography coupled to electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS). A screening level risk assessment combining the measured environmental concentrations (MECs) with dose-response data based on predicted no-effect concentrations (PNECs) was also applied to estimate Hazard Quotients (HQs) for the pharmaceuticals detected in the investigated area. A total of twelve pharmaceuticals were found in the water samples analyzed, with detection frequencies above 78% and in most cases of 100%. Overall, the most abundant pharmaceuticals in surface water were the analgesic and anti-inflammatory drugs naproxen (732-4880ng/L), acetaminophen (354-4460ng/L), and diclofenac (258-1398ng/L), and the lipid regulator bezafibrate (286-2100ng/L). On the contrary, other compounds like the β-blocker atenolol and the psychiatric drug carbamazepine were found at only a few ng or tens of ng per liter in the Apatlaco River. Despite the fact that some of the most abundant compounds showed good removal (>97%) during wastewater treatment, concentrations downstream the WWTP were only slightly lower than upstream. This indicates the existence of additional inputs of untreated wastewater into the river. Based on the obtained HQ-values, the concentrations of ibuprofen, sulfamethoxazole, diclofenac and naproxen present in the river could pose a high toxicity risk for the aquatic ecosystem. These findings highlight these pharmaceuticals as relevant organic contaminants in the area of study and the need to further monitor them in order to adopt appropriate measures to safeguard the ecosystem, and eventually human health.


Retroperitoneal seminoma with simultaneous occurrence in the prostate.
We report a case of retroperitoneal seminoma with simultaneous occurrence in the prostate. The testes were normal on physical and ultrasound examination. In this case a multicentric occurrence of seminoma was considered rather than metastasis from the retroperitoneum to the prostate. Systemic chemotherapy with vincristine, peplomycin and cisplatin was effective for the prostatic as well as the retroperitoneal lesion.


Feasibility of Large-Scale Identification of Sessile Serrated Polyp Patients Using Electronic Records: A Utah Study.
The serrated pathway accounts for 15-25% of sporadic colorectal cancer (CRC). In our study, we sought to accurately characterize sessile serrated polyps (SSP) in a population by electronically interrogating colonoscopy patients' endoscopy and pathology reports using a rules-based text search of pre-defined SSP-related terms. To this aim, we compared a sample of putative SSP and hyperplastic polyps (HP) using our algorithm to a determination of SSP or HP by pathologist and molecular examination to determine the feasibility of large-scale identification of SSP in electronic medical records. In 23,990 endoscopy reports from colonoscopies with pathology performed at a University of Utah Healthcare facility in 2000-2012, we identified serrated lesions and categorized each as putative SSP or HP using a text search algorithm. We obtained 93 tissue samples for histologic and molecular analysis. Serrated polyps were categorized as putative SSP (N = 920) and putative HP (N = 7159) by text search algorithm. Histologic examination of 93 samples identified 37 SSP, 11 probable SSP, and 45 HP. Of 26 putative SSP, 25 were SSP/probable SSP (96%) by histology. Of 67 putative HP, 44 were HP (66%) by histology. Reducing size criterion from ≥1 to ≥5 mm in the search algorithm caused improved sensitivity (77.1%) without decline in specificity (97.8%). A simple rules-based search to identify SSP provides "proof of principle" that SSP can be identified in a large electronic record set. Pilot data indicate defining large, right-sided polyps as ≥5 mm provides adequate sensitivity to detect SSP from electronic records while maintaining high specificity.


Fasting-sensitive SUMO-switch on Prox1 controls hepatic cholesterol metabolism.
Accumulation of excess nutrients hampers proper liver function and is linked to nonalcoholic fatty liver disease (NAFLD) in obesity. However, the signals responsible for an impaired adaptation of hepatocytes to obesogenic dietary cues remain still largely unknown. Post-translational modification by the small ubiquitin-like modifier (SUMO) allows for a dynamic regulation of numerous processes including transcriptional reprogramming. We demonstrate that specific SUMOylation of transcription factor Prox1 represents a nutrient-sensitive determinant of hepatic fasting metabolism. Prox1 is highly SUMOylated on lysine 556 in the liver of ad libitum and refed mice, while this modification is abolished upon fasting. In the context of diet-induced obesity, Prox1 SUMOylation becomes less sensitive to fasting cues. The hepatocyte-selective knock-in of a SUMOylation-deficient Prox1 mutant into mice fed a high-fat/high-fructose diet leads to a reduction of systemic cholesterol levels, associated with the induction of liver bile acid detoxifying pathways during fasting. The generation of tools to maintain the nutrient-sensitive SUMO-switch on Prox1 may thus contribute to the development of "fasting-based" approaches for the preservation of metabolic health.


Evidence for N-methyl-D-aspartate receptor mediation of cocaine induced corticosterone release and cocaine conditioned stimulant effects.
The role of the N-methyl-D-aspartate (NMDA) receptors in cocaine conditioning and sensitization of locomotor activity was studied in four groups of Sprague-Dawley rats. A sub-motoric dose of the NMDA antagonist MK-801 (0.1 mg/kg, i.p.) was employed using a novel dual-compartment Pavlovian drug conditioning paradigm. The animals were placed sequentially in two different test environments in which locomotor activity was monitored. In the first compartment, the animals always received a non-drug test for 20 min. Upon completion of this test, the animals received either saline, cocaine (10 mg/kg i.p.), MK-801 or MK-801 plus cocaine depending on group assignment and were then placed immediately into the second compartment and again tested for 20 min. A total of six non-drug and six drug tests were conducted every other day over a 12-day period. Across all drug/saline treatment and post-treatment tests for conditioning, there were no statistical differences in locomotor activity among the saline and drug treatment groups in the non-drug test environment. In the drug/saline associated environment, however, cocaine had a reliable stimulant effect on locomotion when administered alone or in combination with MK-801. Following a 1-day and again after 21-days of withdrawal, all animals were administered a non-drug test for conditioning in which no injections were administered. On both tests, all groups had equivalent activity levels in the non-drug environment. In the drug/saline environment, only the cocaine group of the three drug treatment groups exhibited conditioned hyperlocomotion. Importantly, MK-801 blocked conditioned hyperlocomotion in the combined cocaine+MK-801 group. MK-801 did not alter serum or brain cocaine concentration or the cocaine effects on dopamine metabolism in limbic brain tissue. The co-administration of MK-801 with cocaine, however, blocked the corticosterone release effect of cocaine. Thus, the NMDA receptor site appears critical for cocaine induced conditioning and for corticosterone release.


[Contractile capacity of fibroblasts from different sources in the model of living skin equivalent].
Human fibroblasts contract collagen gel in vitro and produce a connective tissue-like structure termed the living skin equivalent. In this study, the contractile capacity of postnatal dermal fibroblasts, bone marrow mesenchymal cells and mesenchymal cells derived from the fat tissue has been compared to that of fetal dermal fibroblasts in the model of living skin equivalent. The results show that fetal fibroblasts contract the collagen gel approximately six times stronger than do all other fibroblast cell types, with the numbers of all these cells being equal. A deeper insight into the behavior of fibroblasts differing in their origin will help to develop new approaches to the treatment and regulation of wound healing and fibrosis formation.


Temperature-controlled under-water egg dormancy and postflood hatching in Isotoma viridis (Collembola) as forms of adaptation to annual long-term flooding.
Incubation experiments with eggs of a population of Isotoma viridis, which is exposed to annual long-term flooding from about April to July, as well as field observations show that temperature controls both, underwater egg dormancy and immediate postflood hatching. The population is located at the Eder Freshwater Reservoir in Germany.If constant experimental temperatures are above 14°C, almost all eggs are nondormant. Dormancy is established at temperatures below 15°C, but embryonic development is completed. Experiments indicate that of the environmental factors that change drastically at the end of submergence (light, turgor pressure, oxygen, a.o.), only temperature acts as a hatching trigger. Hatching of the previously dormant eggs occurs at a constant threshold temperature of 16°C, mainly within 2 to 20 days after temperature elevation, but most of these eggs need even higher temperatures to hatch. Remaining eggs were partly stimulated to hatch by recooling them at 7°C for some days and then rewarming them again.The threshold temperatures observed are unusually high for Collembola and seem to be the result of selection by the special floodplain conditions. During normal years, the surface temperatures of submerged soil usually do not exceed threshold limits before summer drainage. This allows both, protection from under-water hatching and an optimal timing of hatching at the very beginning of the main terrestrial period. The experiments show that above the threshold temperature (in warm summers), individuals can hatch under water and survive submerged for 10-15 days. They can survive even longer in the water habitat, if emergent structures enable them to climb onto the water surface. Furthermore, a considerable polymorphism observed in some hatching properties improves the chance to survive under the unpredictable floodplain conditions.


Routine reporting of grey-white matter differentiation in early brain computed tomography in comatose patients after cardiac arrest: A substudy of the COACT trial.
A multimodal approach is advised for neurological prognostication in comatose patients after out-of-hospital cardiac arrest (OHCA). Grey-white matter differentiation (grey-white ratio, GWR) obtained from a brain CT scan performed < 24 hours after return of circulation can be part of this approach. The aims of this study were to investigate the frequency and method of reporting the GWR in brain CT scan reports and their association with outcome. This is a post-hoc descriptive analysis of the COACT trial. The primary endpoint was the reporting of GWR by the radiologist. Secondary endpoints were APACHE IV score, Cerebral Performance Categories at discharge and 90-day follow-up, Glasgow Coma Scale at discharge, GWR-stratified 1-year survival, and RAND-36 stratified by normal versus abnormal GWR. Associations were analysed using multivariable analysis. A total of 427 OHCA patients were included in this study, 234 (55%) of whom underwent a brain CT scan within 24 hours after ROSC. Median time between arrest and initial CT scan was 12 hours. In 195 patients (83%), the GWR was described in the reports, but always expressed qualitatively. The GWR was deemed abnormal in 57 (29%) CT scans. No differences were found in secondary endpoints between the two groups. GWR was frequently described in CT scan reports. Early abnormal GWR, as assessed qualitatively by a radiologist within 24 hours after ROSC, was a poor predictor of neurological prognosis.


Lipofuscin in keratinocytes: Production, properties, and consequences of the photosensitization with visible light.
A dysfunction in the mitochondrial-lysosomal axis of cellular homeostasis is proposed to cause cells to age quicker and to accumulate lipofuscin. Typical protocols to mediate lipofuscinogenesis are based on the induction of the senescent phenotype either by allowing many consecutive cycles of cell division or by treating cells with physical/chemical agents such as ultraviolet (UV) light or hydrogen peroxide. Due to a direct connection with the physiopathology of age-related macular degeneration, lipofuscin that accumulates in retinal pigment epithelium (RPE) cells have been extensively studied, and the photochemical properties of RPE lipofuscin are considered as standard for this pigment. Yet, many other tissues such as the brain and the skin may prompt lipofuscinogenesis, and the properties of lipofuscin granules accumulated in these tissues are not necessarily the same as those of RPE lipofuscin. Here, we present a light-induced protocol that accelerates cell aging as judged by the maximization of lipofuscinogenesis. Photosensitization of cells previously incubated with nanomolar concentrations of 1,9-dimethyl methylene blue (DMMB), severely and specifically damages mitochondria and lysosomes, leading to a lipofuscin-related senescent phenotype. By applying this protocol in human immortalized non-malignant keratinocytes (HaCaT) cells, we observed a 2.5-fold higher level of lipofuscin accumulation compared to the level of lipofuscin accumulation in cells treated with a typical UV protocol. Lipofuscin accumulated in keratinocytes exhibited the typical red light emission, with excitation maximum in the blue wavelength region (~450 nm). Fluorescence lifetime image microscopy data showed that the keratinocyte lipofuscin has an emission lifetime of ~1.7 ns. Lipofuscin-loaded cells (but not control cells) generated a substantial amount of singlet oxygen (1O2) when irradiated with blue light (420 nm), but there was no 1O2 generation when excitation was performed with a green light (532 nm). These characteristics were compared with those of RPE cells, considering that keratinocyte lipofuscin lacks the bisretinoids derivatives present in RPE lipofuscin. Additionally, we showed that lipofuscin-loaded keratinocytes irradiated with visible light presented critical DNA damages, such as double-strand breaks and Fpg-sensitive sites. We propose that the DMMB protocol is an efficient way to disturb the mitochondrial-lysosomal axis of cellular homeostasis, and consequently, it can be used to accelerate aging and to induce lipofuscinogenesis. We also discuss the consequences of the lipofuscin-induced genotoxicity of visible light in keratinocytes.


Amiodarone therapy: role of early and late electrophysiologic studies.
Forty-two patients with a history of symptomatic ventricular tachycardia or cardiac arrest underwent electrophysiologic testing at control and early in the course of amiodarone therapy (mean 12 +/- 7 days). Late electrophysiologic studies (mean 17 +/- 4 weeks) were repeated in 23 patients on a maintenance dose of 400 mg/day. At control study, all patients had inducible ventricular tachyarrhythmias (sustained ventricular tachycardia in 35, nonsustained ventricular tachycardia in 4, ventricular fibrillation in 3), while after amiodarone loading (1,200 mg daily) 4 (10.5%) of the 42 patients developed noninducible ventricular arrhythmias. At late study, an additional 6 (26%) of the 23 patients with inducible arrhythmias at early study developed noninducible arrhythmias. The cycle length of induced ventricular tachycardia increased from 275 +/- 61 ms at control study to 340 +/- 58 ms at early study (p = 0.001). A further increase in ventricular tachycardia cycle length was noted in patients who underwent both early and late study (341 +/- 38 versus 375 +/- 63 ms, p less than 0.05). The percent of induced tachycardias that were clinically tolerated increased as patients were treated longer with amiodarone (control = 22%, early = 34%, late = 53%, p less than 0.001). Of the 23 patients who had both early and late electrophysiologic studies and were followed up for a mean of 21.7 months (range 4 to 47), there were no recurrences among the 6 patients with noninducible arrhythmias, but there were five recurrences among the 17 patients with persistently inducible arrhythmias. None of the four patients with noninducible arrhythmias at early study had a recurrence. On the basis of these findings, it is concluded that: 1) The timing of programmed electrical stimulation will affect the results of the study in patients treated with oral amiodarone.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparative analysis of detoxification-related gene superfamilies across five hemipteran species.
Hemiptera is one of the most speciose orders of insects, and the most speciose considering Hemimetabola. Through their evolutive history, hemipterans with different feeding habits have adapted to deal with different chemical challenges. Three major gene families are involved in xenobiotic detoxification in insects: the cytochromes P450 (CYPs), carboxyl/cholinesterases (CCEs), and glutathione transferases (GSTs). Here we perform a comparative analysis on the complement of these gene superfamilies across five hemipteran species; four heteropterans (the pentatomid plant feeders Nezara viridula and Halyomorpha halys; the hematophagous Cimex lectularius, Cimicidae, and Rhodnius prolixus, Reduviidae), and one Auchenorrhyncha plant feeder (Nilaparvata lugens). Our results point to an expansion of several enzyme families associated with xenobiotic detoxification in heteropterans with respect to other species and the existence of a dynamic evolution pattern including CYP3 clan, hormone and pheromone processing class in the CCE superfamily, and sigma class in GST superfamily. Other detoxification-related families are reduced in the hemipteran species analyzed here: reduction or even absence of epsilon class and reduced delta class in GST superfamily; absence of mitochondrial CYP12 family; absence of CYP9 family in CYP3 clan; and reduction or even absence of some dietary/detoxification groups of CCEs. Interestingly, the most polyphagous species analyzed here (H. halys) is also the one that presents the largest repertoire of detoxification enzymes. Gene cluster analysis suggests that this could be due to gene duplication events. The evolutionary analysis performed here reveals characteristics that are both common and particular for heteropterans. The composition and organization of detoxification-related gene families could shed light on evolutionary forces that shaped their divergence. These families are important for both the detoxification of diet products and for conferring tolerance or resistance to synthetic insecticides. Furthermore, we present the first comprehensive analysis of detoxification gene superfamilies in N. viridula, an understudied species in spite of its economic relevance as a crop pest. The information obtained is of interest for basic insect science as well as for the control of harmful species and the management of insecticide resistance.


Locally advanced breast cancer treated with neutron beams: long-term follow-up in 28 patients.
Between 1972 and 1978, 28 patients with locally advanced breast cancer were treated, 15 with neutron beams only and 13 with mixed neutron and photon beams. Half the patients had inflammatory cancer. For neutrons only, doses ranged between 13.35-25.34 nGy. In mixed-beam regimens, the prescribed total dose ranged between 62 and 76 Gy photon equivalent. Nine patients (32%) had a complete response without local recurrence for the duration of their survival ranging from 1 to 14+ years; 18 patients had a partial response (64%); and one patient had no change. Late toxicity was high: of 24 patients who received tangential breast irradiation, 5 (21%) had ulceration of the breast or chest wall, or both. In four patients, mastectomy and skin grafts were necessary for repair. In only one patient did the skin necrosis heal without corrective surgery. Twelve patients received axillary neutron irradiation, resulting in severe edema in four patients, and brachial plexopathy in six patients. Radiation-induced complications progressed steadily for the duration of the patients' survival after the neutron irradiation. The high complication rate encountered is attributed to high doses resulting from an under estimation of the relative biological effect of the neutron beam for late effects, and to the poor physical and geometrical characteristics of the neutron beam.


What are the causes of non-tolerance to new spectacles and how can they be avoided?
To investigate non-tolerance cases from several UK practices to determine their likely causes and how they might have been avoided. Patient complaint and refraction data were collected from non-tolerance recheck examinations. For one practice, clinical data were also collected retrospectively to investigate the quality of the eye examinations. Data for 279 rechecks were gathered from 10 practices and a recheck frequency of 2.3% was found. The mean patient age was 60 (SD 16) years, with cylinder changes responsible for 38% of prescription-related causes of rechecks, overplusing or underminusing 26%, and underplusing or overminusing just 11%. An assessment of 242 recheck corrections found that 40% were unsatisfactory (e.g., failed to address initial or recheck symptoms, N = 45) and retrospective analysis of 217 case records showed many limitations (e.g., 61% or 28% recorded no uncorrected or habitual visual acuity (VA) at either initial examination or recheck). Given that overplus-underminus was a much bigger proportion of prescription-related cases than overminus-underplus (26% vs. 11%), the refraction mantra of "maximum plus for maximum VA" should be balanced by increased teaching of the problems of overplusing and underminusing, and the use of prescribing guidelines. In addition, continuing professional development regarding the basics of the recheck examination, refraction, visual acuity and prism determination is needed. Changes of oblique cylinders should be carefully considered in older patients as this is a common cause of non-tolerance. In addition, if the "if it ain't broke, don't fix it" and related maxims had been applied to all patients who were asymptomatic at the original examination, one third of all non-tolerance cases could have been avoided. Finally, it would seem appropriate for practices to develop a system to deal better with non-tolerance cases. Perhaps an experienced clinician should examine all patients with non-tolerance and provide feedback to the original clinician.


Time-Driven Activity-Based Costing Provides a Lower and More Accurate Assessment of Costs in the Field of Orthopaedic Surgery Compared With Traditional Accounting Methods.
To analyze the implementation and benefits of time-driven activity-based costing (TDABC) in the field of orthopaedic surgery. We performed a search of PubMed, Google Scholar, and Embase in March 2020, using the following terms: "Time-Driven Activity-Based Costing," "TDABC," "Orthopaedic Surgery," and "Cost." Then we selected the studies that used the TDABC methodology to generate costs for a particular aspect of orthopaedic surgery. The included studies were divided into the following 5 main categories for ease of analysis: joint arthroplasty, trauma, hand, electronic medical record (EMR) implementation, and pediatric. We analyzed the overall ability of TDABC in the field of orthopaedic surgery, compared to the standard costing methods. We included a total of 19 studies that implemented the TDABC methodology to generate a cost, which was compared to traditional accounting methods. The orthopaedic subspecialty with the most amount of TDABC implementation has been the field of joint arthroplasty. In these studies, the authors have noted that TDABC has provided a more granular breakdown of costs and has calculated a lower cost compared with traditional accounting methods. TDABC is a powerful cost analysis method that has demonstrated benefit over the activity-based costing (ABC) approach in determining a lower and more accurate cost of orthopaedic procedures. Furthermore, the TDABC method generates an average cost reduction of $10,000 and $12,000 for total hip arthroplasty and total knee arthroplasty, respectively. TDABC can allow health care administration to better determine and understand the cost drivers of particular orthopaedic procedures at their institutions. With improved estimates on the true cost of an activity, hospital administrators and department chairs can adjust to ensure cost-effective, patient-centered health care.


Insulin-stimulated protein kinase activity in rat skeletal muscle that phosphorylates ribosomal protein S6.
Treatment of rats with a single high dose of insulin leads to rapid stimulation of cytosolic protein kinase activity in skeletal muscle that phosphorylates ribosomal protein S6. This stimulation is maximal within 15 minutes after insulin treatment, and the activity remains elevated for at least 90 minutes. The insulin-stimulated protein kinase activity elutes as two peaks from DEAE-Sepharose. Peak I elutes at 0.04-0.06 M KCl and is stimulated by insulin approximately 1.4-fold above the control. Peak II elutes at 0.09-0.11 M KCl and is stimulated 2.8-fold above the control. The peak II activity, which is most strongly stimulated by insulin, is resolved from cyclic AMP-dependent protein kinase on DEAE-Sepharose and appears to be distinct from protein kinase C. These results represent a novel finding of the stimulation of S6 kinase activity by insulin in skeletal muscle tissue in vivo.


Thermostable direct hemolysin diminishes tyrosine phosphorylation of epidermal growth factor receptor through protein kinase C dependent mechanism.
Adequate evidence mounts to the fact that several bacteria and their toxins have protective or curative roles in colorectal cancers. Thermostable direct hemolysin (TDH), produced by Vibrio parahaemolyticus, down regulates cell proliferation in colon carcinoma cell lines. TDH induces Ca2+ influx from an extracellular environment accompanied by protein kinase C phosphorylation. Activated protein kinase C inhibits the tyrosine kinase activity of epidermal growth factor receptor (EGFR), the rational target of anti-colorectal cancer therapy. Immunoblotting analyses were performed to ascertain protein kinase C activation, EGFR status, EGFR phosphorylation and mitogen activated protein kinase (MAPK) activity. Flow cytometry analysis and ELISA reconfirmed tyrosine phosphorylation of EGFR and ERK activations, respectively. PKC-α siRNA knockdown was done to corroborate the involvement of PKC-α in the undertaken study. Our study showed the translocation of PKC-α from cytosol to the membrane fraction in colon carcinoma cell lines on incubation with TDH. The EGFR tyrosine kinase activity exhibited a down regulation on TDH treatment which involved PKC-α, as confirmed by siRNA knockdown. Also ERK phosphorylation occurred on PKC-α activation. TDH activated PKC-α down regulates EGFR tyrosine kinase activity by MEK dependent mechanism involving MAPK. In this study we have seen that TDH has an implication in EGFR based therapeutic approach in colorectal cancer via PKC mediated mechanism.


Prebiotics and Probiotics: Effects on Dyslipidemia and NAFLD/NASH and the Associated Mechanisms of Action.
Non-alcoholic fatty liver disease (NAFLD) is now considered the most common chronic liver disease worldwide. NAFLD is related to changes in lipid metabolism and is characterized by the increase or accumulation of fat in hepatocytes that may progress to nonalcoholic steatohepatitis (NASH), which leads to the appearance of inflammatory processes. Treatment consists of changes in diet, physical activity, and weight control; however, these disorders represent a health problem and require the development of novel alternatives to treatment and prevention. NAFLD/NASH are strongly associated with other disorders, such as metabolic syndrome (MetS); in fact, NAFLD is considered the hepatic manifestation of MetS. These disorders are related to other components of MetS, including dyslipidemia, which is characterized by an imbalance in blood cholesterol and triglyceride levels. Prebiotics and probiotics benefit from treating and preventing several ailments, including liver diseases. Specifically, in dyslipidemia, NAFLD, and NASH, probiotics play a fundamental role in conducting the biotransformation of primary bile acids into secondary bile acids, which generally have important activity as immunomodulators and metabolism regulators. The mechanisms of action of pre and probiotics involve the activity of bile acid receptors, such as FXR and TGR-5, and the events resulting from their activation. Therefore, prebiotics and probiotics may be reasonable options to prevent and treat metabolic- related liver diseases.


Modified high-performance liquid chromatographic determination of diamine oxidase activity in plasma.
A previous high-performance liquid chromatographic determination of diamine oxidase activity suitable for tissue homogenates was modified in order to adapt it to plasma samples. Simple additional steps were introduced after the enzyme reaction and before the chromatographic separation, both according to the previous method. In this way the sensitivity and the reproducibility of the overall procedure was suitable for routine plasma diamine oxidase estimations.


Bilingualism protects anterior temporal lobe integrity in aging.
Cerebral gray-matter volume (GMV) decreases in normal aging but the extent of the decrease may be experience-dependent. Bilingualism may be one protective factor and in this article we examine its potential protective effect on GMV in a region that shows strong age-related decreases-the left anterior temporal pole. This region is held to function as a conceptual hub and might be expected to be a target of plastic changes in bilingual speakers because of the requirement for these speakers to store and differentiate lexical concepts in 2 languages to guide speech production and comprehension processes. In a whole brain comparison of bilingual speakers (n = 23) and monolingual speakers (n = 23), regressing out confounding factors, we find more extensive age-related decreases in GMV in the monolingual brain and significantly increased GMV in left temporal pole for bilingual speakers. Consistent with a specific neuroprotective effect of bilingualism, region of interest analyses showed a significant positive correlation between naming performance in the second language and GMV in this region. The effect appears to be bilateral though because there was a nonsignificantly different effect of naming performance on GMV in the right temporal pole. Our data emphasize the vulnerability of the temporal pole to normal aging and the value of bilingualism as both a general and specific protective factor to GMV decreases in healthy aging.


TLD skin dose measurements and acute and late effects after lumpectomy and high-dose-rate brachytherapy only for early breast cancer.
This report examines the relationships between measured skin doses and the acute and late skin and soft tissue changes in a pilot study of lumpectomy and high-dose-rate brachytherapy only for breast cancer. Thirty-seven of 39 women enrolled in this pilot study of high-dose-rate brachytherapy (37.2 Gy in 10 fractions b.i.d.) each had thermoluminescent dosimetry (TLD) at 5 points on the skin of the breast overlying the implant volume. Skin changes at TLD dose points and fibrosis at the lumpectomy site were documented every 6 to 12 months posttreatment using a standardized physician-rated cosmesis questionnaire. The relationships between TLD dose and acute skin reaction, pigmentation, or telangiectasia at 5 years were analyzed using the GEE algorithm and the GENMOD procedure in the SAS statistical package. Fisher's exact test was used to determine whether there were any significant associations between acute skin reaction and late pigmentation or telangiectasia or between the volumes encompassed by various isodoses and fibrosis or fat necrosis. The median TLD dose per fraction (185 dose points) multiplied by 10 was 9.2 Gy. In all 37 patients, acute skin reaction Grade 1 or higher was observed at 5.9% (6 of 102) of dose points receiving 10 Gy or less vs. 44.6% (37 of 83) of dose points receiving more than 10 Gy (p < 0.0001). In 25 patients at 60 months, 1.5% telangiectasia was seen at dose points receiving 10 Gy or less (1 of 69) vs. 18% (10 of 56) telangiectasia at dose points receiving more than 10 Gy (p = 0.004). Grade 1 or more pigmentation developed at 1.5% (1 of 69) of dose points receiving less than 10 Gy vs. 25% (14 of 56) of dose points receiving more than 10 Gy (p < 0.001). A Grade 1 or more acute skin reaction was also significantly associated with development of Grade 1 or more pigmentation or telangiectasia at 60 months. This association was most significant for acute reaction and telangiectasia directly over the lumpectomy site (p < 0.001). Grade 1 or more fibrosis, in 25 patients with a 60-month follow-up, occurred in 47.4% (9 of 19) of patients with a volume of 45 cm3 or less covered by the 100% isodose vs. 83.3% (5 of 6) of patients with a larger volume (p = 0.180). Asymptomatic and biopsy-proven fat necrosis occurred in 5 patients. No significant differences in fat necrosis rates according to volume were detected. For high-dose-rate brachytherapy to the lumpectomy site, TLD skin dose was significantly related to acute skin reaction and to pigmentation and telangiectasia at 60 months. An acute skin reaction was also significantly associated with the development of telangiectasia at 60 months. TLD skin dose measurement may allow modification of the brachytherapy implant geometry (dwell times and position) to minimize late skin toxicity.


Illu-Shoal Choice: An Exploration of Different Means for Enrichment of Captive Zebrafish.
Fish of any variety are nowadays being kept captive for several purposes, from recreational to alimentary to research. It is possible that we humans often underestimate or misunderstand the basic, natural needs of the species we use for our purposes. Sociality is likely to play an extensive and fundamental role in the quality of life of animals such as zebrafish. This study aimed to develop a dummy conspecific that included depth and motion illusions in order to assess whether these stimuli could represent a valid alternative to a conspecific in triggering shoaling behaviour in a well-known model in genetics and neuroscience, the zebrafish (Dario rerio). We thus replaced the natural livery of a zebrafish shape with three visual illusions: the Ouchi-Spillmann illusion, which generates an effect of local tilting motion; and another two which should create pictorial cues of tridimensionality. Via a binary shoal choice test, we assessed the time spent close to each of the three artificial dummies compared to neutral control stimuli such as grey ellipses. We found no preference for the illusory patterns, suggesting that the illusion was not perceived or, alternatively, that the perception of the illusion was not enough to elicit recognition of the dummy as conspecific and subsequent social behaviours.


Photodynamic therapy for human hepatoma-cell-line tumors utilizing biliary excretion properties of indocyanine green.
Photodynamic therapy (PDT) has not been reported for human hepatoma, because cancer cells only weakly take up the photosensitizer. Indocyanine green (ICG) is a photosensitizer normally excreted into the bile, and bile excretion is impaired in human hepatomas. We examined whether human hepatoma cell lines preferentially take up the ICG and then assessed the effectiveness of PDT using ICG and near-infrared (NIR) laser. HuH-7 and HepG2 human hepatoma cell lines were transplanted subcutaneously into mice. Developing HuH-7 and HepG2 tumors were confirmed that preferentially took up the ICG in 24 h after ICG was administered to mice via tail vein. The HuH-7 tumor showed a high tumor-to-background fluorescence intensity ratio, 255:1, whereas fluorescence intensity of HuH-7 is increased twofold compared to HepG2. HuH-7 cell transplanted mice were divided into three groups: ICG administration only (ICG+NIR-, n = 8), ICG and NIR laser exposure (ICG+NIR+, n = 12), and NIR laser exposure only (ICG-NIR+, n = 5). Mean tumor volume in the ICG+NIR- and ICG-NIR+ groups increased steadily. In contrast, mean tumor volume in the ICG+NIR+ group did not change between days 0 and 3. Mean tumor volume did not differ significantly between the ICG-NIR+ and ICG-NIR- groups, but was significantly different between the ICG+NIR+ group and both the ICG-NIR+ and ICG+NIR- groups (p < 0.01). ICG is preferentially taken up by HuH-7 and HepG2 human hepatoma cell line tumors. The tumor-to-background ratio of HuH-7 tumors, in particular, was extremely high. PDT with NIR laser irradiation suppressed HuH-7 human hepatoma cell line tumor growth.


Side dependent effects of the human amnion on angiogenesis.
Amnion (AM), the innermost layer of human placenta, has a variety of functions such as capability to reduce scarring and inflammation, as well as anti-microbial and immunoregulatory properties. However, there are challenging reports about angiogenic and anti-angiogenic effects of the AM. The aim of this study was to evaluate whether the angiogenesis is dependent on epithelial or mesenchymal sides of this membrane. Dorsal skinfold chamber model was performed on male rats. A layer of dorsal skin of rats was removed and the AM was implanted in either epithelial side up or mesenchymal side up position. Intra-vital microscopy was done one week after tissue transplantation. In vitro evaluation of angiogenesis was also performed using rat aortic ring assay on the AM. The number of vessel sprouts and their lengths were increased more significantly in epithelial side up group comparing to the control group. Inhibitory effect of epithelial side of the AM on angiogenesis was clearly seen in mesenchymal side up group. Both number and length of sprouts in mesenchymal up group were decreased in comparison to epithelial side up group. In aortic ring assay, angiogenesis was detected on the AM after removal of the amniotic epithelial cells. This study showed that the AM has both angiogenic and anti-angiogenic properties, which is surface dependent. Therefore, the AM can have a vast application in both ischemic organs through inducing angiogenesis and pathological situations such as cancer in which angiogenesis must be inhibited.


Nonpenetrating femtosecond laser intrastromal astigmatic keratotomy in eyes having cataract surgery.
To describe the effect of femtosecond laser intrastromal astigmatic keratotomy (AK) performed during cataract surgery. Moorfields Eye Hospital, London, United Kingdom. Prospective case series. This study comprised patients having laser cataract surgery with concurrent astigmatism management by intrastromal AK. All eyes had greater than 0.7 corneal diopter (D) cylinder. An intrastromal AK nomogram with 8.0 mm diameter paired symmetric limbal centered arcs was used. Corneal keratometry was measured preoperatively and 1 month postoperatively using a KR8100PA topographer-autorefractor. Astigmatic analyses were performed using the Alpins method considering 3 vectors-target induced astigmatism (TIA), surgically induced astigmatism (SIA) and difference vector (DV)-and calculation of coupling measures. In all, 196 eyes of 133 patients were analyzed. The mean TIA (equivalent to preoperative corneal cylinder) was 1.21 D ± 0.42 (SD) (range 0.75 to 2.64 D) and the mean SIA was 0.74 DC ± 0.40 (range 0.00 to 2.86). The mean difference vector was 0.74 ± 0.38 D (range 0.00 to 2.25 D). The mean correction index was 0.63 ± 0.32 (range 0.00 to 1.93), indicating that the mean astigmatism correction was 63%. Fourteen eyes (7.1%) and 7 eyes (3.6%) had an astigmatism correction of greater than 100% and greater than 120%, respectively. Overall 0%, 48.5%, and 51.5% of eyes had 0.50 D or less, 1.0 D or less, or greater than 1.0 D, respectively, preoperatively compared with 32.1%, 85.7%, and 14.3%, postoperatively. There were no cases of corneal endothelial perforation or inadvertent placement within the visual axis. The intrastromal AKs were easily programmed as an integral part of laser-assisted cataract surgery without additional cost, significantly reduced corneal cylinder, and appeared to be safe through 1 month of follow-up. Dr. Day was supported by the National Institute for Health Research (NIHR) Biomedical Research Centre based at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology. Dr. Stevens has been a consultant to Optimedica Inc., now part of Abbott Medical Optics.


Full evaporation dynamic headspace and gas chromatography-mass spectrometry for uniform enrichment of odor compounds in aqueous samples.
A method for analysis of a wide range of odor compounds in aqueous samples at sub-ng mL⁻¹ to μg mL⁻¹ levels was developed by full evaporation dynamic headspace (FEDHS) and gas chromatography-mass spectrometry (GC-MS). Compared to conventional DHS and headspace solid phase microextraction (HS-SPME), FEDHS provides more uniform enrichment over the entire polarity range for odor compounds in aqueous samples. FEDHS at 80°C using 3 L of purge gas allows complete vaporization of 100 μL of an aqueous sample, and trapping and drying it in an adsorbent packed tube, while providing high recoveries (85-103%) of the 18 model odor compounds (water solubility at 25°C: log0.54-5.65 mg L⁻¹, vapor pressure at 25°C: 0.011-3.2 mm Hg) and leaving most of the low volatile matrix behind. The FEDHS-GC-MS method showed good linearity (r²>0.9909) and high sensitivity (limit of detection: 0.21-5.2 ng mL⁻¹) for the model compounds even with the scan mode in the conventional MS. The feasibility and benefit of the method was demonstrated with analyses of key odor compounds including hydrophilic and less volatile characteristics in beverages (whiskey and green tea). In a single malt whiskey sample, phenolic compounds including vanillin could be determined in the range of 0.92-5.1 μg mL⁻¹ (RSD<7.4%, n=6). For a Japanese green tea sample, 48 compounds including 19 potent odorants were positively identified from only 100 μL of sample. Heat-induced artifact formation for potent odorants was also examined and the proposed method does not affect the additional formation of thermally generated compounds. Eighteen compounds including 12 potent odorants (e.g. coumarin, furaneol, indole, maltol, and pyrazine congeners) were determined in the range of 0.21-110 ng mL⁻¹ (RSD<10%, n=6).


Purification of aquaculture wastewater by macrophytes and biofilm systems: Efficient removal of trace antibiotics and enrichment of antibiotic resistance genes.
The purification performance of aquaculture wastewater and the risk of antibiotic resistance genes (ARGs) dissemination in wetlands dominated by macrophytes remain unclear. Here, the purification effects of different macrophytes and biofilm systems on real aquaculture wastewater were investigated, as well as the distribution and abundance of ARGs. Compared to the submerged macrophytes, artificial macrophytes exhibited higher removal rates of TOC (58.80 ± 5.04 %), TN (74.50 ± 2.50 %), and TP (77.33 ± 11.66 %), and achieved approximately 79.92 % removal of accumulated trace antibiotics in the surrounding water. Additionally, the biofilm microbial communities on the surface of artificial macrophytes exhibited higher microbial diversity with fewer antibiotic-resistant bacteria (ARB) enrichment from the surrounding water. The absolute abundance of ARGs (sul1, sul2, and intI1) in the mature biofilm to be one to two orders of magnitude higher than that in the water. Although biofilms could decrease ARGs in the surrounding water by enriching ARB, the intricate network structure of biofilms further facilitated the proliferation of ARB and the dissemination of ARGs in water. Network analysis suggested that Proteobacteria and Firmicutes phyla were dominant and potential carriers of ARGs, contributing 69.00 % and 16.70 %, respectively. Our findings highlight that macrophytes and biofilm systems have great performance on aquaculture wastewater purification, but with high risk of ARGs.


Preoperative prognostic factors associated with poor early recovery after trapeziectomy with ligament reconstruction and tendon interposition arthroplasty for thumb carpometacarpal osteoarthritis.
Trapeziectomy with ligament reconstruction and tendon interposition (LRTI) is performed for treating thumb carpometacarpal osteoarthritis. However, patients occasionally experience delayed postoperative recovery. Although several prognostic factors associated with long-term outcomes have been identified, the factors associated with delayed recovery after trapeziectomy with LRTI have not been identified. When we define major disability in the early postoperative period at 3 months after LRTI surgery as delayed recovery, some preoperative factors can influence with delayed recovery. Moreover, delayed recovery can influence the long-term therapeutic outcome. Thirty thumbs that underwent trapeziectomy with LRTI (mean follow-up period, 29.3 months) were retrospectively analyzed. Major disability was defined by both DASH and Hand20 scores≥35 after surgery, and patients were divided into groups 1 (major disability; n=9) and 2 (no major disability; n=21) according to the scores at 3 months after surgery. Preoperative data, including age, sex, preoperative symptom duration, dominant hand surgery, concurrent surgery with LRTI, previous orthopedic surgery, employment, stage of osteoarthritis, thickness of the trapezium, metacarpophalangeal hyperextension deformity, DASH score, Hand20 score, visual analogue scale (VAS) scores for pain, grip strength, pinch strength, and range of motion of the thumb were compared between groups to identify the factors indicating a poor prognosis at 3 months after surgery. We also compared the clinical outcomes at the 12-month and final follow-up. Dominant hand surgery, previous orthopedic surgery, and preoperative poor DASH score were significantly more frequent in group 1, while the other factors did not show significant intergroup differences. Moreover, group 1 showed significantly poorer DASH score, grip and pinch strength, and active flexion of the thumb metacarpal joint at both the 12-month and final follow-up. This group also showed significantly poorer Hand20 and VAS scores at the final follow-up. Dominant hand surgery, previous orthopedic surgery, and preoperative poor DASH score were associated with poor recovery at 3 months after trapeziectomy with LRTI. Moreover, major disability at 3 months after surgery influenced poor clinical outcomes at the 12-month and final follow-ups. These data could be useful for counseling patients regarding the expected recovery duration and outcomes after LRTI surgery. IV; retrospective therapeutic study.


Possible consequences of applying guidelines to healthy women with a family history of breast cancer.
Possible effects of consistently applying published guidelines on healthy women with breast cancer in their family history were analysed. We investigated 1060 unrelated breast cancer patients and calculated the numbers of first-degree relatives that would be referred to a familial cancer clinic if the guidelines were consistently applied. A first-degree relative was considered a candidate for referral if she was female, without breast cancer at the moment of the interview, alive and over the age of 24. The criteria for referral were based on one Dutch and two British guidelines. According to the Dutch guideline, for one affected woman with breast cancer, 0.25 (95% CI 0.22-0.28) healthy first-degree female relatives should be offered a consultation at a familial cancer clinic (FCC). Application of the British guidelines would lead to a similar number of referrals. Of all healthy first-degree female relatives, who should be referred to an FCC, 34-37% had an index case among their family who was already known at a genetic department. If current guidelines are consistently applied, a sharp increase in referrals to FCCs may be expected. These guidelines, however, are arbitrary and only limited data are available on the efficacy of this surveillance for high-risk healthy women.


Coordinate induction of multiple mRNAs specific for rat liver phenobarbital-inducible cytochromes P-450.
Utilizing two-dimensional gel electrophoresis, the polypeptide composition of a purified microsomal cytochrome P-450 preparation isolated from phenobarbital-treated Long-Evans rats obtained from Charles River Laboratories has been examined. The purified protein consists of three polypeptides with nearly identical subunit molecular weights (approximately 52,000) but differing in net charge. These three polypeptides can be detected in liver microsomes isolated from phenobarbital-treated rats by immunoblot analysis but are virtually absent in microsomes isolated from untreated rats. All three polypeptides appear to be products of distinct mRNAs since they can be immunoprecipitated from rabbit reticulocyte lysates programmed with poly(A+)-RNA isolated from phenobarbital-treated rats. The amount of functional mRNA specific for the P-450 polypeptides increases dramatically in response to an acute administration of phenobarbital; however, in untreated rats the amount of functional mRNA was below the level of detection by the translational assay. These data are consistent with the very low level of the phenobarbital-inducible cytochromes P-450 in liver microsomes isolated from untreated rats. Finally, the data indicate that all three cytochrome P-450 mRNAs increase rapidly in response to phenobarbital administration and are regulated coordinately.


Evaluation of reading comprehension with neuropsychological and event-related brain potential (ERP) methods.
Evidence is presented for the efficacy of a new method of assessing reading comprehension using a standardized reading test that was formatted for computer presentation with simultaneous event-related brain potential (ERP) recordings. Reading comprehension abilities of 23 healthy undergraduate students were evaluated using ERPs. The results revealed a differential ERP response pattern for correct and incorrect test items. These response patterns were observed at individual participant levels. The findings provided further support for the use of ERPs in the neuropsychological assessment of patients who are difficult or impossible to assess using behavioral responses because of concomitant motoric and/or communicative limitations.


Comparison Between Radiological Versus Endoscopic Assessment of Adenoid Tissue in Patients of Chronic Adenoiditis.
The adenoids are a common cause for morbidity in pre-school and school going children. Adenoid hypertrophy is common cause of nasal obstruction in pediatric population. The assessment of the size of palatine tonsil hypertrophy is quiet easy to perform, however, measuring the size of nasopharyngeal tonsil and establishing its connection with the given symptoms is more difficult. This is basically due to the location of a tonsil in the nasopharyngeal cavity. Since the cavity is inaccessible directly and cannot be examined visually, many direct and indirect methods of adenoid size evaluation have been developed. The objective of this study is to compare the diagnostic efficacy of both X-ray soft tissue nasopharynx lateral view and nasoendoscopy in evaluation of adenoid size in patients having chronic adenoiditis. Our study included 50 children in age group of 3-14 years who had signs and symptoms of adenoiditis clinically. After comparing X-ray and endoscopic findings we found that there was good agreement between X-ray and endoscopic method. Sensitivity of X-ray was 79.41% and specificity of 75% whereas sensitivity of nasoendoscopy was 87.10% and specificity was 63.16%. Nasal endoscopy is good screening method and more accurate method to assess the size of adenoids in patients of chronic adenoiditis.