Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2.
The gene CHEK2 encodes a checkpoint kinase playing a key role in the DNA damage pathway. Though CHEK2 has been identified as an intermediate breast cancer susceptibility gene, only a small proportion of high-risk families have been explained by genetic variants located in its coding region. Alteration in gene expression regulation provides a potential mechanism for generating disease susceptibility. The detection of differential allelic expression (DAE) represents a sensitive assay to direct the search for a functional sequence variant within the transcriptional regulatory elements of a candidate gene. We aimed to assess whether CHEK2 was subject to DAE in lymphoblastoid cell lines (LCLs) from high-risk breast cancer patients for whom no mutation in BRCA1 or BRCA2 had been identified. We implemented an assay based on high-resolution melting (HRM) curve analysis and developed an analysis tool for DAE assessment. We observed allelic expression imbalance in 4 of the 41 LCLs examined. All four were carriers of the truncating mutation 1100delC. We confirmed previous findings that this mutation induces non-sense mediated mRNA decay. In our series, we ruled out the possibility of a functional sequence variant located in the promoter region or in a regulatory element of CHEK2 that would lead to DAE in the transcriptional regulatory milieu of freely proliferating LCLs. Our results support that HRM is a sensitive and accurate method for DAE assessment. This approach would be of great interest for high-throughput mutation screening projects aiming to identify genes carrying functional regulatory polymorphisms.


Antitumor Activity of Portulaca Oleracea L. Polysaccharide on HeLa Cells Through Inducing TLR4/NF-κB Signaling.
Abstarct We have previously shown that Portulaca oleracea L. polysaccharide (POL-P3b) possesses the ability to inhibit cervical cancer cell growth in vitro and in vivo. In this study, we explored how toll-like receptor 4 (TLR4) signaling correlated with the antitumor mechanism of POL-P3b. Western blotting was utilized to detect the expression of TLR4 and the downstream signaling pathway. The level of inflammatory mediator was quantified using enzyme-linked immunosorbent assay (ELISA) kits. The effects of POL-P3b on the proliferation and apoptosis in HeLa cells were determined by WST-8 assay and Hoechst 33342/propidium iodide (PI) assay. Our results demonstrated that lipopolysaccharide (LPS) binding to TLR4 on tumor cells could enhance HeLa cell proliferation and increase the expression of TLR4 and the downstream molecules. Treating HeLa cells with POL-P3b could decrease the proliferation of HeLa cells, and upregulate Bax level and downregulate Bcl-2 level in a concentration-dependent manner. In addition, POL-P3b inhibited the protein expression levels of TLR4, MyD88, TRAF6, Activator Protein-1 (AP-1) and nuclear factor-κB (NF-κB) subunit P65 in HeLa cells. Furthermore, POL-P3b also reduced the production of cytokine/chemokine. Taken together, the present work suggested the antitumor mechanism of POL-P3b by downregulating TLR4 downstream signaling pathway and inducing cell apoptosis. Our results may provide direct evidence to suggest that POL-P3b should be considered as a potent nutrient supplement for oncotherapy.


Ultrasonography of the Achilles tendon in hypercholesterolemia.
The ultrasonographic (US) appearance of Achilles tendon was examined in 34 patients with hypercholesterolemia (10 with familial (FH), and 24 with secondary hypercholesterolemia (SH)), and in 22 patients with normal tendons. US findings were compared with clinical, laboratory, and radiologic results. In patients with FH, typical xanthomas in the form of hyperechoic tendinal tumors were found in only 15% while various forms of inhomogeneity of tendon structure without xanthoma formation were found in 75%. The high rate of tendon inhomogeneity may be due to the concomitant occurrence of both xanthomas and degeneration of tendon fibers. Physical examination revealed abnormalities in 60% of these patients. At CT of the tendons, abnormalities were found in 65% of the FH patients and in 40%, abnormalities were shown by plain radiography. In SH, the results did not differ significantly from normal controls. In our opinion US should be used to prove or rule out Achilles tendon abnormalities in patients with FH for prophylaxis and treatment of tendinitis and tendon rupture.


The association of FAD with the cytochrome b-245 of human neutrophils.
A plasma membrane fraction prepared from human neutrophils had a fluorescence resembling that of a fluorescent flavoprotein, with emission maximum near 520nm and excitation maxima near 380 and 460nm. The fluorescence emission and excitation properties of Triton N-101-solubilized membrane fraction resembled those of FAD. FAD was present in the membranes at a concentration of 417pmol/mg of protein and cytochrome b(-245) at a concentration of 407pmol/mg of protein. In a 110-fold purified preparation of cytochrome b(-245) the ratio of FAD:cytochrome b was 1:1. Analytical gradient centrifugation of neutrophil homogenates shows a coincidence of two cytochrome b peaks and two peaks of fluorescence, corresponding with plasma membrane and specific granule fractions; most of the FAD was non-fluorescent and located in fractions lighter than the plasma membrane. Plasma membrane fractions prepared from neutrophils of patients suffering from the X-linked form of chronic granulomatous disease lacked cytochrome b and contained 194pmol of FAD/mg of protein; plasma membrane fractions prepared from neutrophils of patients with the autosomal recessive form of chronic granulomatous disease contained both cytochrome b(-245) and FAD in the normal range of concentrations in a ratio of 1:1. Phagocytic vesicles were prepared from normal neutrophils and found to contain FAD and cytochrome b in a ratio 2.22:1, suggesting that activation of neutrophils many involve the incorporation of an additional flavin into the membrane. Under anaerobic conditions in the presence of EDTA to act as an electron donor to a flavin, the cytochrome b(-245) of neutrophil membranes was partly (12%) photoreducible, an effect increased to 100% by the addition of FMN. The extent of reduction of cytochrome b in an anaerobic neutrophil homogenate containing NADH increased from 30% to 70% on illumination. We suggest that these results indicate a close association between FAD and cytochrome b(-245) and support a scheme for electron transport thus: [Formula: see text]


Immune Hemolysis: Diagnosis and Treatment Recommendations.
Autoimmune hemolytic anemia (AIHA) is a heterogeneous disease usually classified as warm, cold [cold agglutinin disease (CAD)] or mixed, according to the thermal range of the autoantibody. Diagnosis is based on the direct antiglobulin test (DAT), typically positive with anti-IgG antisera in warm AIHA and anti-C3d in CAD. Diagnostic pitfalls are due to IgA autoantibodies, warm IgM, low-affinity IgG, or IgG below the threshold of sensitivity, and about 5% of AIHA remains DAT-negative. The treatment of AIHA is still not evidence-based. Corticosteroids are the first-line therapy for warm AIHA. For refractory/relapsed cases, the choice is between splenectomy (effective in ~70% cases but with a presumed cure rate of 20%) and rituximab (effective in ~70%-80% of cases), which is becoming the preferred second-line treatment, and thereafter any of the immunosuppressive drugs (azathioprine, cyclophosphamide, cyclosporine, mycophenolate mofetil). Rituximab is now recommended as first-line treatment for CAD. Additional therapies are intravenous immunoglobulins, danazol, and plasma exchange, with alemtuzumab and high-dose cyclophosphamide as the last options.


Data independent acquisition-digital archiving mass spectrometry: application to single kernel mycotoxin analysis of Fusarium graminearum infected maize.
New and conjugated mycotoxins of concern to regulators are frequently being identified, necessitating the costly need for new method development and sample reanalysis. In response, we developed an LC-data independent acquisition (LC-DIA) method on a Q-Exactive Orbitrap mass spectrometer tailored for mycotoxins analysis. This method combines absolute quantification of targeted fungal metabolites with non-targeted digital archiving (DA) of data on all ionizable compounds for retrospective analysis. The quantitative power of this approach was assessed by spiking 23 mycotoxins at a range of concentrations into clean maize extracts. The linearity and limits of detection achieved were comparable to conventional LC-MS/MS and significantly better than 'all-ion-fragmentation' scanning mode. This method was applied to single kernel analysis of Fusarium infected maize, where we quantified nine Fusarium metabolites and three metabolites from unexpected contaminations by Alternaria and Penicillium species. Retrospective analysis of this data set allowed us to detect the recently reported 15-acetyldeoxynivalenol-3-O-β-D-glucoside without requiring re-analysis of the samples. To our knowledge, this is the first reported occurrence of this conjugated mycotoxin in naturally contaminated maize, and led us to further study maize artificially inoculated with the 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol chemotypes of Fusarium graminearum. Analysis of these samples showed that the maize genotype tested glycosylates 15-acetyldeoxynivalenol but not 3-acetyldeoxynivalenol likely because the glycosylation site was blocked. In addition to confirming that these two F. graminearum chemotypes behave differently when infecting the host plant, it demonstrates the utility of using a single screening method to quantify known mycotoxins and archive a completely non-targeted dataset for future analysis.


Effects of blood donation on cerebral blood flow velocity.
Eighteen healthy male blood donors, nine with hematocrit (Hct) of 0.40 to 0.45 (normal Hct) and nine with Hct of 0.49 to 0.52 (upper-limit Hct), were monitored by continuous-wave internal carotid Doppler sonography and hematologic tests for 28 days after blood donation, to ascertain whether and to what extent a single standard donation may modify the velocity of cerebral blood flow. The two groups had similar mean predonation values of internal carotid flow velocity (ICFV): blood donation was followed in both groups by a slight, transient decrease of ICFV at the end of phlebotomy, due to donation-induced hypovolemia, and then by an increase of ICFV lasting 7 to 10 days. Analysis of individual profiles revealed that only four of nine upper-limit and six of nine normal Hct donors displayed a positive trend (increase) in the ICFV within the first week after donation, and that it was due mainly to a rise in systolic flow velocity. Mean Hct and arterial oxygen content showed a negative trend (decrease) within the first week that was opposite to the ICFV trend. Other laboratory variables, including serum proteins and plasma fibrinogen concentration, and the iron status indicators did not change, except for serum ferritin, which also decreased within the first week after phlebotomy. It can be concluded that blood donation may result in a short-term increase of blood flow velocity that is independent of Hct predonation levels in approximately one-half of the donors.


Relationships between musical structure and psychophysiological measures of emotion.
Psychophysiological studies with music have not examined what exactly in the music might be responsible for the observed physiological phenomena. The authors explored the relationships between 11 structural features of 16 musical excerpts and both self-reports of felt pleasantness and arousal and different physiological measures (respiration, skin conductance, heart rate). Overall, the relationships between musical features and experienced emotions corresponded well with those known between musical structure and perceived emotions. This suggests that the internal structure of the music played a primary role in the induction of the emotions in comparison to extramusical factors. Mode, harmonic complexity, and rhythmic articulation best differentiated between negative and positive valence, whereas tempo, accentuation, and rhythmic articulation best discriminated high arousal from low arousal. Tempo, accentuation, and rhythmic articulation were the features that most strongly correlated with physiological measures. Music that induced faster breathing and higher minute ventilation, skin conductance, and heart rate was fast, accentuated, and staccato. This finding corroborates the contention that rhythmic aspects are the major determinants of physiological responses to music.


Recognition of the epidermal growth factor receptor by reovirus.
The demonstration that alpha-sialic acid is the minimal determinant recognized by human reovirus is compatible with the finding that this virus binds to multiple sialoglycoproteins on the host cell surface. However, the identities of these proteins have remained unknown. By applying detergent-solubilized plasma membranes from the human epidermoid carcinoma A431 cell line to immobilized reovirions, we have identified the 150- to 170-kDa epidermal growth factor (EGF) receptor as one of the cell surface proteins recognized by reovirus. Direct interaction between the N-terminal extracellular domain of the EGF receptor and reovirus was confirmed by the demonstration that of a number of proteins secreted by A431 cells, the 105-kDa N-terminal cell surface domain of the EGF receptor was the major protein recognized by the virus. However, as expected, reovirus and EGF did not compete for the same binding site on the EGF receptor of intact A431 cells.


Association of Monoamine Oxidase A (MAOA) Gene uVNTR and rs6323 Polymorphisms with Attention Deficit and Hyperactivity Disorder in Korean Children.
Objective: Attention deficit hyperactivity disorder (ADHD) is a common neurodevelopmental disorder. The genetic cause of ADHD is still unclear, but the dopaminergic, serotonergic, and noradrenergic pathways have shown a strong association. In particular, monoamine oxidase A (MAOA) plays an important role in the catabolism of these neurotransmitters, suggesting that the MAOA gene is associated with ADHD. Therefore, we evaluated the relationship between the MAOA gene polymorphisms (uVNTR and rs6323) and ADHD. Materials and methods: We collected a total of 472 Korean children (150 ADHD cases and 322 controls) using the Korean version of the Dupaul Attention Deficit Hyperactivity Disorder Rating Scales (K-ARS). Genotyping was performed by PCR and PCR-RFLP. The Behavior Assessment System for Children Second Edition (BASC-2) was used to evaluate the problem behaviors within ADHD children. Results: We observed significant associations between the rs6323 and ADHD in girls (p < 0.05) and the TT genotype was observed as a protective factor against ADHD in the recessive model (OR 0.31, 95% CI 0.100⁻0.950, p = 0.022). The 3.5R-G haplotype showed a significant association in ADHD boys (p = 0.043). The analysis of subtype also revealed that the 4.5R allele of uVNTR was a risk factor for the development of ADHD in the combined symptom among girls (OR 1.87, 95% CI 1.014⁻3.453, p = 0.031). In the BASC-2 analysis, the MAOA uVNTR polymorphism was associated with activities of daily living in ADHD boys (p = 0.017). Conclusion: These results suggest the importance of the MAOA gene polymorphisms in the development of ADHD in Korean children. A larger sample set and functional studies are required to further elucidate of our findings.


Human hematopoietic microenvironments.
Dormancy of hematopoietic stem cells and formation of progenitors are directed by signals that come from the bone marrow microenvironment. Considerable knowledge has been gained on the murine hematopoietic stem cell microenvironment, while less so on the murine progenitor microenvironment and even less so on these microenvironments in humans. Characterization of these microenvironments is decisive for understanding hematopoiesis and finding new treatment modalities against bone marrow malignancies in the clinic. However, it is equally challenging, because hematopoietic stem cells are difficult to detect in the complex bone marrow landscape. In the present study we are characterizing the human hematopoietic stem cell and progenitor microenvironment. We obtained three adjacent bone marrow sections from ten healthy volunteers. One was used to identify a population of CD34+/CD38- "hematopoietic stem cells and multipotent progenitors" and a population of CD34+/CD38+ "progenitors" based on immunofluorescence pattern/intensity and cellular morphology. The other two were immunostained respectively for CD34/CD56 and for CD34/SMA. Using the combined information we performed a non-computer-assisted quantification of nine bone marrow components (adipocytes, megakaryocytes, bone surfaces, four different vessel types (arteries, capillaries, sinusoids and collecting sinuses), other "hematopoietic stem cells and multipotent progenitors" and other "progenitors") within 30 μm of "hematopoietic stem cells and multipotent progenitors", "progenitors", and "random cell profiles". We show that the microenvironment of the "hematopoietic stem cells and multipotent progenitors" is significantly enriched in sinusoids and megakaryocytes, while the microenvironment of the "progenitors" is significantly enriched in capillaries, other "progenitors", bone surfaces and arteries.


Enhanced production of ligninolytic enzymes and decolorization of molasses distillery wastewater by fungi under solid state fermentation.
Selected isolates of fungi were grown on wheat straw and corncob in the presence of different moistening agents such as water, molasses, potato dextrose broth and distillery effluent. All the fungal isolates responded differently with respect to growth and ligninolytic enzyme production. Fungal growth on different substrates was checked by calculating ergosterol content, which varied widely within a single species when grown on different substrates. The maximum laccase production was obtained for Aspergillus flavus TERI DB9 grown on wheat straw with molasses. For manganese peroxidase, highest production was in Aspergillus niger TERI DB20 grown on corncob with effluent. Among the two isolates positive for lignin peroxidase, the highest production was in Fusarium verticillioides ITCC 6140. This immobilized fungal biomass was then used for decolorization of effluent from a cane molasses based distillery. Maximum decolorization (86.33%) was achieved in Pleurotus ostreatus (Florida) Eger EM 1303 immobilized on corncob with molasses in a period of 28 days.


[Use of an enzyme immunoassay test system with cystic Echinococcus antigen to diagnose echinococcosis alveolaris (multilocularis) (alveococcosis)].
Twenty-three serum samples from 12 patients with Echinococcus alveolaris collected 1 to 12 years after surgery and 1--13 courses of specific therapy were used to evaluate the diagnostic effectiveness of an enzyme immunoassay (EIA) test system with cystic Echinococcus antigen. The findings enable the EIA test system with cystic Echinococcus antigen to be recommended for the diagnosis of Echinococcosis alveolaris and for the monitoring of its treatment results.


[Using urinary diagnostic indices to evaluate acute renal failure in epidemic hemorrhagic fever].
The urinary diagnostic indices were used to evaluate acute renal failure in 118 cases with epidemic hemorrhagic fever (EHF). When the renal failure was mainly caused by acute tubular necrosis, it would occur earlier and persist longer. The authors suggested that the indices are helpful in documenting the severity and identifying the characteristics of acute renal failure in EHF. The mechanisms for developing acute renal failure in EHF was discussed.


Case report: the lupus anticoagulant-hypoprothrombinemia syndrome.
The lupus anticoagulant is a well-described in vitro phenomenon that may be associated with arterial and venous thrombotic episodes. The lupus anticoagulant is never accompanied by a hemorrhagic diathesis unless it is associated with a second coagulation abnormality such as thrombocytopenia or hypoprothrombinemia. The lupus anticoagulant-hypoprothrombinemia syndrome is now a well-defined entity that may cause a severe, life-threatening hemorrhagic diathesis. The hypoprothrombinemia in this syndrome is the result of rapid clearance of prothrombin-antiprothrombin antibody complexes by the reticulo-endothelial system. The cause of antiprothrombin antibody formation is unknown. The authors describe a recent experience with a patient with this syndrome who initially had recurrent, life-threatening gastrointestinal bleeding. They were able to demonstrate hypoprothrombinemia and the presence of prothrombin-antiprothrombin antibody immune complexes. The patient was treated with prednisone, with correction of the bleeding disorder; however, the patient had resultant death from thrombosis. A literature review of the past 30 years as it relates to the discovery and treatment of this phenomenon is included.


The Precursor of the Cretaceous-Tertiary Boundary Clays at Stevns Klint, Denmark, and DSDP Hole 465A.
Results of detailed mineralogical, chemical, and oxygen isotope analyses of the clay minerals and zeolites from two Cretaceous-Tertiary (K/T) boundary regions, Stevns Klint, Denmark, and Deep Sea Drilling Project (DSDP) Hole 465A in the north central Pacific Ocean, are presented. In the central part of the Stevns Klint K/T boundary layer, the only clay mineral detected by x-ray diffraction is a pure smectite with > 95 percent expandable layers. No detrital clay minerals or quartz were observed in the clay size fraction in these beds, whereas the clay minerals above and below the boundary layer are illite and mixed-layer smectite-illite of detrital origin as well as quartz. The mineralogical purity of the clay fraction, the presence of smectite only at the boundary, and the delta(18)O value of the smectite (27.2 +/- 0.2 per mil) suggest that it formed in situ by alteration of glass. Formation from impact rather than from volcanic glass is supported by its major element chemistry. The high content of iridium and other siderophile elements is not due to the cessation of calcium carbonate deposition and resulting slow sedimentation rates. At DSDP Hole 465A, the principal clay mineral in the boundary zone (80 to 143 centimeters) is a mixed-layer smectite-illite with >/=90 percent expandable layers, accompanied by some detrital quartz and small amounts of a euhedral authigenic zeolite (clinoptilolite). The mixed-layer smectite-illite from the interval 118 to 120 centimeters in the zone of high iridium abundance has a very low rare earth element content; the negative cerium anomaly indicates formation in the marine environment. This conclusion is corroborated by the delta(18)O value of this clay mineral (27.1 +/- 0.2 per mil). Thus, this mixed-layer smectite-illite formed possibly from the same glass as the K/T boundary smectite at Stevns Klint, Denmark.


[The therapeutic posture in desmoid tumor].
The desmoid tumor is a histologically benign neoplasm originating in muscular-aponeurotic structures and possesses a high tendency of local recurrence. The aggressive nature of the tumor obliges radical surgery to be performed principally in extra-abdominal tumors. Twelve patients were treated from 1983 to 1993. The treatments performed prior to and after histologic diagnosis, the time of evolution when radical treatment was carried out upon diagnosis and the evolution after treatment were evaluated. Of the patients with desmoid tumor 8 were males with a mean age of 29.9 +/- 12.5 years. In eleven cases the presentation was as a tumor and was of pain in only one case. All the patients were surgically treated with a total of 27 interventions being carried out. Three patients also received pharmacologic treatment with tamoxifen, in 2 cases associated to sulindac. In two radiotherapy was used but not as a first line treatment. Following a mean follow up of 38 +/- 17 months no mortality has been registered but 5 patients are not disease free. The therapeutic attitude to the desmoid tumor should be radical surgery in both the primary tumor and the recurrences with medical treatment being performed in specific situations.


Activation of NF-kappaB by extracellular matrix is involved in spreading and glucose-stimulated insulin secretion of pancreatic beta cells.
Laminin-5-rich extracellular matrix derived from 804G cells (804G-ECM) engages beta1 integrins to induce spreading, improve glucose-stimulated insulin secretion (GSIS), and increase survival of pancreatic beta cells. The present study examines whether 804G-ECM activates the transcriptional activity of NF-kappaB and the involvement of NF-kappaB in those effects of 804G-ECM on pancreatic beta cells. 804G-ECM induces nuclear translocation and the DNA binding activity of the p65 subunit of NF-kappaB. 804G-ECM-induced nuclear translocation of NF-kappaB was weak as compared with that induced by interleukin-1beta. Transient 804G-ECM-induced DNA binding activity of NF-kappaB (peak at 2 h) and overexpression of NF-kappaB target genes IkappaB alpha and NF-kappaB1(p105) (peak at 4 h) were observed. When NF-kappaB was inhibited by an inhibitor of IkappaB alpha phosphorylation (Bay 11-7082) or by a recombinant adenovirus expressing the nonphosphorylatable form of IkappaB alpha, 804G-ECM-induced cell spreading and actin cytoskeleton organization were reduced. GSIS from cells on 804G-ECM was inhibited 5-fold, whereas cell survival was not affected. In summary, the results indicate that 804G-ECM induces a transient and moderate NF-kappaB activity. This study shows for the first time that ECM-induced NF-kappaB activity is necessary in maintaining GSIS, although it does not affect survival of pancreatic beta cells. The effects of ECM-induced NF-kappaB activity contrast with the deleterious effects of cytokine-induced NF-kappaB activity. It is proposed that transient and moderate NF-kappaB activity is essential for proper function of the pancreatic beta cell.


Production and characterization of bio-oils from fast pyrolysis of tobacco processing wastes in an ablative reactor under vacuum.
Application of advanced pyrolysis processes to agricultural waste for liquid production is gaining great attention, especially when it is applied to an economic crop like tobacco. In this work, tobacco residues were pyrolyzed in an ablative reactor under vacuum. The maximum bio-oil yield of 55% w/w was obtained at 600°C with a particle size of 10 mm at a blade rotation speed of 10 rpm. The physical properties of the products showed that the oil produced was of high quality with high carbon, hydrogen, and calorific value. Two-dimensional gas chromatography/time-of-flight mass spectrometric analysis results indicated that the oils were complex mixtures of alkanes, benzene derivative groups, and nitrogen-containing compounds. In addition, 13C NMR results confirmed that long aliphatic chain alkanes were evident. The alkanes were likely converted from furans that were decomposed from hemicelluloses. Ablative pyrolysis under vacuum proved to be a promising option for generating useful amount of bio-oils from tobacco residues.


Microarray image segmentation using additional dye--an experimental study.
The DNA microarray technique allows monitoring the expression levels of thousands of genes simultaneously. A single DNA microarray experiment involves a number of error-prone manual and automated processes, which influence the results and have an impact on the subsequent stages of analysis. Typical problems of arrays are pinning errors while probe printing and the corruption of spots by noise patches. These errors should be detected at the time of image analysis in order to prevent the erroneous intensities from ending up in the analysis and inference stages. In this paper we introduce the concept (referred to as SybrSpot) of utilizing information provided by an additional dye, SYBR green RNA II, for segmentation of gene expression microarrays. Owing to the effective binding of the SYBR green RNA II to the array probes, an image with high signal-to-noise ratio is obtained. This image is used to learn about the spot quality and to flag spots which are not reliably hybridized and corrupted by noise. Further, we compare SybrSpot with GenePix and demonstrate that SybrSpot performs better than GenePix when flagging spots with no probes or weak probes. The code is available upon request to authors.


Realities of automating OPA HPLC amino acid analyses.
The levels of free amino acids in culture medium can easily be monitored by reversed-phase HPLC chromatography on a C18 column following pre-column derivatization with o-pthaladehyde. The method does not require the prior removal of proteins. A guard column does that. When this method was automated, problems started to occur after about eight injections. Peak resolution rapidly degraded. Unidentified matter was retained by the column and produced both interfering peaks and high back pressures. Better resolution of the closely eluting amino acids occurred when both the pH of the loading buffer was increased and the gradient time was lengthened. Special washes and a carefully controlled time of equilibration with the loading buffer helped to minimize back pressure and interfering substances. Other often skipped routine maintenance and solvent handling procedures were also necessary for optimal performance. These modifications made routine runs of 40 plus samples possible as well as dramatically increased the lifetime of the column.


On-line detection of radioactive and non-radioactive heavy metals in tobacco smoke using portable laser-induced breakdown spectroscopy.
Tobacco smoke is a complex mixture consisting of hundreds of toxic and carcinogenic chemicals, which may cause many diseases once inhaled. Among these toxic substances, radioactive and non-radioactive heavy metals present great health concerns to both active and passive smokers. However, conventional methodologies for characterizing heavy metals in smoke require intensive and time-consuming preparation procedures, and thus prevent their on-line applications. We report here a portable laser-induced breakdown spectroscopy (PLIBS) device for on-line detection of radioactive and non-radioactive heavy metals in tobacco smoke with ultra-high sensitivity. Two local cigarette brands were investigated, and a number of heavy metals were detected. In particular, several radioactive metals show very strong atomic emissions, and a limit of detection down to parts per trillion was obtained by single-shot PLIBS. The linear dependencies of their spectral intensities on the smoke concentrations were revealed, suggesting that the PLIBS is a promising tool for high-throughput and quantitative analysis of heavy metals in tobacco smoke. Tobacco emission dynamics were also investigated, and the results suggest the potential applications of the PLIBS in smoke risk assessment.


Persistent solar influence on North Atlantic climate during the Holocene.
Surface winds and surface ocean hydrography in the subpolar North Atlantic appear to have been influenced by variations in solar output through the entire Holocene. The evidence comes from a close correlation between inferred changes in production rates of the cosmogenic nuclides carbon-14 and beryllium-10 and centennial to millennial time scale changes in proxies of drift ice measured in deep-sea sediment cores. A solar forcing mechanism therefore may underlie at least the Holocene segment of the North Atlantic's "1500-year" cycle. The surface hydrographic changes may have affected production of North Atlantic Deep Water, potentially providing an additional mechanism for amplifying the solar signals and transmitting them globally.


Air pollution and symptoms of depression in elderly adults.
Although the effect of air pollution on various diseases has been extensively investigated, few studies have examined its effect on depression. We investigated the effect of air pollution on symptoms of depression in an elderly population. We enrolled 537 participants in the study who regularly visited a community center for the elderly located in Seoul, Korea. The Korean version of the Geriatric Depression Scale-Short Form (SGDS-K) was used to evaluate depressive symptomatology during a 3-year follow-up study. We associated ambient air pollutants with SGDS-K results using generalized estimating equations (GEE). We also conducted a factor analysis with items on the SGDS-K to determine which symptoms were associated with air pollution. SGDS-K scores were positively associated with interquartile range (IQR) increases in the 3-day moving average concentration of particulate matter with an aerodynamic diameter ≤ 10 μm (PM10) [17.0% increase in SGDS-K score, 95% confidence interval (CI): 4.9%, 30.5%], the 0-7 day moving average of nitrogen dioxide [NO2; 32.8% (95% CI: 12.6%, 56.6%)], and the 3-day moving average of ozone [O3; 43.7% (95% CI: 11.5%, 85.2%)]. For these three pollutants, factor analysis showed that air pollution was more strongly associated with emotional symptoms such as feeling happy and satisfied than with somatic or affective symptoms. Our study suggests that increases in PM10, NO2, and O3 may increase depressive symptoms among the elderly. Of the symptoms evaluated, ambient air pollution was most strongly associated with emotional symptoms.


English-Language Proficiency, Academic Networks, and Academic Performance of Mexican American Baccalaureate Nursing Students.
We examine how academic networks mediate between English-language proficiency and academic performance. The relationship between English-language proficiency and academic performance remains inconclusive; it is possible that academic networks play a role in this relationship. Filling this knowledge gap is central to building best practices in teaching, and to evaluating the impact of networks on success. Data were analyzed from 164 Mexican American nursing students. We used English Language Acculturation Scale (ELAS) items as predictors, interaction with academic networks as the mediating variable, and course grade as the outcome; regression analyses were performed. Interaction with academic networks correlated with grades; ELAS was not significant. Instead, academic networks mediated between entrance GPA and grades, an unexpected finding. Academic networks are critical in academic performance. However, only those students who have a history of high performance are likely to have or to activate academic networks.


CD99 triggering in Ewing sarcoma delivers a lethal signal through p53 pathway reactivation and cooperates with doxorubicin.
The paucity of new drugs for the treatment of Ewing sarcoma (EWS) limits the cure of these patients. CD99 has a strong membranous expression in EWS cells and, being also necessary for tumor survival, is a suitable target to aim at. In this article, we described a novel human monospecific bivalent single-chain fragment variable diabody (dAbd C7) directed against CD99 of potential clinical application. In vitro and in vivo evaluation of cell death and of the molecular mechanisms triggered by anti-CD99 agents were performed alone or in combination with doxorubicin to demonstrate efficacy and selectivity of the new dAbd C7. The dAbd C7 induced rapid and massive EWS cell death through Mdm2 degradation and p53 reactivation. Mdm2 overexpression as well as silencing of p53 in p53wt EWS cells decreased CD99-induced EWS cell death, whereas treatment with nutlin-3 enhanced it. Furthermore, cell death was associated with induction of p21, bax, and mitochondrial depolarization together with substantial inhibition of tumor cell proliferation. Combined treatment of anti-CD99 dAbd C7 with doxorubicin was additive both in vitro and in vivo against EWS xenografts. Normal mesenchymal stem cells showed no p53 activation and were resistant to cell death, unless transformed by EWS-FLI, the oncogenic driver of EWS. These results indicate that dAbd C7 is a suitable candidate tool to target CD99 in patients with EWS able to spare normal stem cells from death as it needs an aberrant genetic context for the efficient delivery of CD99-triggered cell death.


Exposure to domestic violence in women living in Istanbul and Aegean regions: a Turkish sample.
Domestic violence (DV) is a serious public health problem in the world. DV against women is also a global problem without cultural, geographic, religious, social, economic or national boundaries. This descriptive cross-sectional study was carried out to determine the situations of DV in women living in Istanbul and the Aegean Region in Turkey. The study population included outpatient clinics of state hospitals both regions. A stratified sampling by age was performed and 1100 women were included into the sample. Data were collected at face-to-face interviews with Domestic Violence Against Women Determination Scale. The mean age of the women living in Istanbul was 41.81 ± 9.75 years and Aegean Region was 33.72 ± 11.38 years. The prevalence of emotional and financial violence were higher in Istanbul and the Aegean Region. The women living in Istanbul got higher scores for Domestic Violence Against Women Determination Scale. The prevalence of the women reporting to suffer from violence from their spouses was 15.4% in Istanbul and 14% in the Aegean Region. While the prevalence of the women suffering from violence was higher in Istanbul, the women in Aegean Region suffered from more severe violence. The violence prevalence was lower among the wives and the husbands with high education levels, employed women and high-income families.


Dislocation-enhanced experimental-scale vacancy loop formation in hcp Zirconium in one single collision cascade.
Large defects are the main factor leading to the degradation of material properties under irradiation environments. It is commonly assumed that the large defects are mainly formed through cluster growth under continuous irradiations. Besides this mechanism, recent experiments and simulations show that sometimes an individual ion can also directly create a large defect. Here we report a novel mechanism for the formation of the large defects, as discovered by our Molecular Dynamics (MD) simulations of the collision cascades in hcp Zirconium (Zr): a pre-existing edge dislocation (ED) can significantly promote the nucleation of the vacancy clusters, and even facilitate the direct formation of an experimental-scale large vacancy loop (about 3 nm) in only one single displacement cascade. This dislocation-related mechanism may be the key for understanding the experimental results in the low-dose irradiated Zr where the high-density large dislocation loops are observed but difficult to be explained by the two mechanisms mentioned above. Considering that intrinsic dislocations exist in nearly all crystalline materials, our results provide a significant concept: pre-existing dislocations have a strong influence on the primary damage production, and taking them into account is indispensable for assessing and improving the material's irradiation-resistance.


Abnormalities in cortical pattern of coherence in migraine detected using ultra high-density EEG.
Individuals with migraine generally experience photophobia and/or phonophobia during and between migraine attacks. Many different mechanisms have been postulated to explain these migraine phenomena including abnormal patterns of connectivity across the cortex. The results, however, remain contradictory and there is no clear consensus on the nature of the cortical abnormalities in migraine. Here, we uncover alterations in cortical patterns of coherence (connectivity) in interictal migraineurs during the presentation of visual and auditory stimuli and during rest. We used a high-density EEG system, with 128 customized electrode locations, to compare inter- and intra-hemispheric coherence in the interictal period from 17 individuals with migraine (12 female) and 18 age- and gender-matched healthy control subjects. During presentations of visual (vertical grating pattern) and auditory (modulated tone) stimulation which varied in temporal frequency (4 and 6 Hz), and during rest, participants performed a colour detection task at fixation. Analyses included characterizing the inter- and intra-hemisphere coherence between the scalp EEG channels over 2-s time intervals and over different frequency bands at different spatial distances and spatial clusters. Pearson's correlation coefficients were estimated at zero-lag. Repeated measures analyses-of-variance revealed that, relative to controls, migraineurs exhibited significantly (i) faster colour detection performance, (ii) lower spatial coherence of alpha-band activity, for both inter- and intra-hemisphere connections, and (iii) the reduced coherence occurred predominantly in frontal clusters during both sensory conditions, regardless of the stimulation frequency, as well as during the resting-state. The abnormal patterns of EEG coherence in interictal migraineurs during visual and auditory stimuli, as well as at rest (eyes open), may be associated with the cortical hyper-responsivity that is characteristic of abnormal sensory processing in migraineurs.


Long-Term Outcomes and Responses to Retreatment in Patients With Melanoma Treated With PD-1 Blockade.
To analyze long-term outcomes after treatment discontinuation of anti-programmed death-1 (anti-PD-1) therapy in a cohort of patients with melanoma with the longest follow-up yet available to our knowledge, including a majority of patients treated outside of a clinical trial. We also assessed efficacy of retreatment with anti-PD-1 therapy with or without ipilimumab in relapsing patients. We retrospectively analyzed all patients with nonuveal, unresectable stage III/IV melanoma treated with single-agent anti-PD-1 therapy at Memorial Sloan Kettering from 2009-2018 who had discontinued treatment and had at least 3 months of follow-up after discontinuation (n = 396). Overall survival for patients with complete response (CR) was calculated from time of CR. Time to treatment failure for patients with CR was time from CR to the next melanoma treatment or death. CRs were seen in 102 of 396 patients (25.8%). The median number of months of treatment after CR was zero (range, stopped before CR to 26 months after CR). With a median follow-up of 21.1 months from time of CR in patients who did not relapse, the probability of being alive and not needing additional melanoma therapy at 3 years was 72.1%. There was no significant association between treatment duration and relapse risk. In multivariable analysis, CR was associated with M1b disease and cutaneous versus mucosal or acral primaries. Among the 78 patients (of 396) retreated after disease progression, response was seen in 5 of 34 retreated patients with single-agent anti-PD-1 therapy and 11 of 44 patients escalated to anti-PD-1 plus ipilimumab. In our cohort, most patients discontinued treatment at the time of CR. Most CRs were durable but the probability of treatment failure was 27% at 3 years. Responses to retreatment were infrequent. The optimal duration of treatment after CR is not yet established.


Changes in the pattern of admissions for attempted suicide in Newcastle upon Tyne during the 1960s.
Comparison of 726 admissions to Newcastle upon Tyne General Hospital for attempted suicide in the period 1 October 1966 to 30 September 1969 with a previously reported series of 276 in 1962-4 showed an overall increase in annual admissions of 77%. For both sexes under age 30 years and females over 60 years the increase was over double. Psychiatric morbidity had increased, and there was also a significant rise in the proportion of unemployed men.The relative frequency of ingestion of non-barbiturate psychotropic or hypnotic drugs and barbiturates had changed, the former now being more common than the latter as the suicidal agent. Finally, there had been a significant increase in the proportion of patients receiving subsequent psychiatric treatment.


[Parapapillary diameter of retinal vessels. I. Estimating the size of the optic papilla (a papillometric study of over 264 normal eyes)].
The diameter of the temporal superior and inferior retinal artery and vein were measured at the optic disk border and 2 mm from the disk center in 264 nonselected normal eyes on the basis of photographs, with correction of magnification. Additionally, the individual ocular magnification factor was determined by Littmann's method. The mean caliber of the artery was 0.113 +/- 0.019 mm at the disk border and 0.118 +/- 0.023 2 mm from the disk center. The corresponding values for the vein were 0.149 +/- 0.026 mm and 0.159 +/- 0.026 mm. The correlations between vessel caliber and disk size were not significant, but the relationship between disk size and the quotient of vessel diameter divided by disk size was highly significant (p less than 0.0001). Vessel diameter can therefore be taken as an independent unit for determining optic disk size in relative and approximately also in absolute units. Thus, mean horizontal disk diameter (1.77 +/- 0.33 mm) corresponded to 16.3 +/- 3.5 artery and 12.3 +/- 2.6 vein diameters (measured at the optic disk border). The mean individual ocular magnification factor was 0.302 +/- 0.017, with a Gauss standard distribution. If the optical characteristics of a photographed eye are not known and the ocular magnification factor is assumed to be 0.302, then 68.3% of all eyes will be covered with an inaccuracy of 5.6% and 95.5% of all eyes with an inaccuracy of 11.2%.


Parental risk attitudes and children's academic test scores: evidence from the US panel study of income dynamics.
Using a sequence of questions from the 1996 US Panel Study of Income Dynamics (PSID), we explore the implications of interpersonal differences in parent's attitudes towards risk for the academic test scores of their children focusing on information drawn from the 1997 Child Development Supplement of the PSID. In addition, we explore whether parental risk preference influences whether the child subsequently attends college. Our findings suggest that a parent's degree of risk aversion is inversely related to the academic test scores of their children as well as being inversely related to the probability of attending college post high school.


Progress With Nonhuman Animal Models of Addiction.
Nonhuman animals have been major contributors to the science of the genetics of addiction. Given the explosion of interest in genetics, it is fair to ask, are we making reasonable progress toward our goals with animal models? I will argue that our goals are changing and that overall progress has been steady and seems likely to continue apace. Genetics tools have developed almost incredibly rapidly, enabling both more reductionist and more synthetic or integrative approaches. I believe that these approaches to making progress have been unbalanced in biomedical science, favoring reductionism, particularly in animal genetics. I argue that substantial, novel progress is also likely to come in the other direction, toward synthesis and abstraction. Another area in which future progress with genetic animal models seems poised to contribute more is the reconciliation of human and animal phenotypes, or consilience. The inherent power of the genetic animal models could be more profitably exploited. In the end, animal research has continued to provide novel insights about how genes influence individual differences in addiction risk and consequences. The rules of the genetics game are changing so fast that it is hard to remember how comparatively little we knew even a generation ago. Rather than worry about whether we have been wasting time and resources asking the questions we have been, we should look to the future and see if we can come up with some new ones. The valuable findings from the past will endure, and the sidetracks will be forgotten.


Large loop excision of the transformation zone.
During the early 1990s, the technique of LLETZ has acquired widespread approval throughout much of the colposcopic community, and its advantages over destructive methods of treatment have been described by several authors. 1. It allows for histologic audit of the colposcopic diagnosis. 2. It allows histopathologic examination to rule out microinvasion. 3. It allows excision of the dysplastic lesion and the transformation zone, which may be confirmed histologically. 4. It may be performed at the first (assessment) colposcopic examination. 5. It may be adapted to treat all cases of CIN, irrespective of the size and site of the transformation zone. 6. It is an easily learned technique. 7. It uses inexpensive, readily available equipment and has low operating costs. 8. It is usually an office or outpatient procedure performed using local anesthesia. The last five of these eight advantages are also potential disadvantages of the technique that may combine to increase the morbidity of the procedure. Women may be treated more easily and at a lower threshold of abnormality in the office with local anesthesia and with transformation zones of almost any dimension, situated on the ectocervix in the endocervical canal or both. If more women are treated (at a lower threshold of suspected abnormality), then procedure-related morbidity will increase. Because the technique allows sufficient flexibility to accommodate transformation zones of every site and dimension, it is inevitable that women who would otherwise have had a cone biopsy will now have a LLETZ procedure. The morbidity of a cone biopsy (LLETZ, laser, or cold knife) is related to the volume, and also probably the amount of endocervical tissue excised. It is important that the morbidity associated with removal of a long endocervical transformation zone be recognized as a consequence of the size and site of the transformation zone, rather than of the choice of excisional technique. EASE OF USE The method is technically straightforward and undemanding to an experienced and relatively dexterous colposcopist. COST When compared with the laser technique, the method is less expensive. However, the other destructive modalities are equally inexpensive (cold coagulation, cryocautery, and radical diathermy). Many of the electrosurgical units that are used for LLETZ also may be used for a variety of procedures in gynecology. EFFECTIVENESS It is evident that a method of excising the transformation zone will have the same likelihood of successfully treating women with CIN as do the destructive techniques, and this has been supported by the published series of patients treated by LLETZ. It is also true that LLETZ is unlikely to significantly improve on the success/failure rates of treatment achieved by the protagonists of each destructive method of treatment. This is because the success/failure rates of destructive methods are high when performed by experts. However, women with CIN cannot always be treated by experts of individual destructive techniques. Perhaps a more clinically important question is whether LLETZ is associated with a superior success/failure rate compared with destructive methods in which each are performed by the nonspecialized practicing gynecologist.


HLA Class Ia and Ib Polyreactive Anti-HLA-E IgG2a Monoclonal Antibodies (TFL-006 and TFL-007) Suppress Anti-HLA IgG Production by CD19+ B Cells and Proliferation of CD4+ T Cells While Upregulating Tregs.
The anti-HLA-E IgG2a mAbs, TFL-006 and TFL-007, reacted with all HLA-I antigens, similar to the therapeutic preparations of IVIg. Indeed, IVIg lost its HLA reactivity, when its HLA-E reactivity was adsorbed out. US-FDA approved IVIg to reduce antibodies in autoimmune diseases. But the mechanism underlying IVIg-mediated antibody reduction could not be ascertained due to the presence of other polyclonal antibodies. In spite of it, the cost prohibitive high or low IVIg is administered to patients waiting for donor organ and for allograft recipients for lowering antiallograft antibodies. A mAb that could mimic IVIg in lowering Abs, with defined mechanism of action, would be highly beneficial for patients. Demonstrably, the anti-HLA-E mAbs mimicked several functions of IVIg relevant to suppressing the antiallograft Abs. The mAbs suppressed activated T cells and anti-HLA antibody production by activated B cells, which were dose-wise superior to IVIg. The anti-HLA-E mAb expanded CD4+, CD25+, and Foxp3+ Tregs, which are known to suppress T and B cells involved in antibody production. These defined functions of the anti-HLA-E IgG2a mAbs at a level superior to IVIg encourage developing their humanized version to lower antibodies in allograft recipients, to promote graft survival, and to control autoimmune diseases.


Movement by helicopter of patients with decompression sickness.
Rapid movement of a patient with decompression sickness sometimes poses problems when the site of the hyperbaric treatment facility is located a considerable distance away. Six cases of aviator decompression sickness were diagnosed in altitude chamber participants during an 18-month period. Five cases were uncomplicated decompression sickness and the sixth case was of central nervous system decompression sickness. All cases were transferred by low-level helicopter flight. No complications were noted when the helicopter stayed within 200 ft (61 m) AGL of the take-off point. Symptoms of decompression sickness did worsen however, when this altitude was exceeded. This study shows that movement of patients with decompression sickness by low-level helicopter flight is both safe and effective, especially when pressurized aircraft is neither available nor practical.


Bacterial Competition Systems Share a Domain Required for Inner Membrane Transport of the Bacteriocin Pyocin G from Pseudomonas aeruginosa.
Bacteria exploit a variety of attack strategies to gain dominance within ecological niches. Prominent among these are contact-dependent inhibition (CDI), type VI secretion (T6SS), and bacteriocins. The cytotoxic endpoint of these systems is often the delivery of a nuclease to the cytosol. How such nucleases translocate across the cytoplasmic membrane of Gram-negative bacteria is unknown. Here, we identify a small, conserved, 15-kDa domain, which we refer to as the inner membrane translocation (IMT) domain, that is common to T6SS and bacteriocins and linked to nuclease effector domains. Through fluorescence microscopy assays using intact and spheroplasted cells, we demonstrate that the IMT domain of the Pseudomonas aeruginosa-specific bacteriocin pyocin G (PyoG) is required for import of the toxin nuclease domain to the cytoplasm. We also show that translocation of PyoG into the cytosol is dependent on inner membrane proteins FtsH, a AAA+ATPase/protease, and TonB1, the latter more typically associated with transport of bacteriocins across the outer membrane. Our study reveals that the IMT domain directs the cytotoxic nuclease of PyoG to cross the cytoplasmic membrane and, more broadly, has been adapted for the transport of other toxic nucleases delivered into Gram-negative bacteria by both contact-dependent and contact-independent means. IMPORTANCE Nuclease bacteriocins are potential antimicrobials for the treatment of antibiotic-resistant bacterial infections. While the mechanism of outer membrane translocation is beginning to be understood, the mechanism of inner membrane transport is not known. This study uses PyoG as a model nuclease bacteriocin and defines a conserved domain that is essential for inner membrane translocation and is widespread in other bacterial competition systems. Additionally, the presented data link two membrane proteins, FtsH and TonB1, with inner membrane translocation of PyoG. These findings point to the general importance of this domain to the cellular uptake mechanisms of nucleases delivered by otherwise diverse and distinct bacterial competition systems. The work is also of importance for the design of new protein antibiotics.


Changes in peripheral blood compounds following psychopharmacological treatment in drug-naïve first-episode patients with either schizophrenia or major depressive disorder: a meta-analysis.
This meta-analysis on peripheral blood compounds in drug-naïve first-episode patients with either schizophrenia or major depressive disorder (MDD) examined which compounds change following psychopharmacological treatment. The Embase, PubMed and PsycINFO databases were systematically searched for longitudinal studies reporting measurements of blood compounds in drug-naïve first-episode schizophrenia or MDD. For this random-effects meta-analysis, we retrieved a total of 31 studies comprising 1818 schizophrenia patients, and 14 studies comprising 469 MDD patients. Brain-derived neurotrophic factor (BDNF) increased following treatment in schizophrenia (Hedges' g (g): 0.55; 95% confidence interval (CI) 0.39-0.70; p < 0.001) and MDD (g: 0.51; CI 0.06-0.96; p = 0.027). Interleukin (IL)-6 levels decreased in schizophrenia (g: -0.48; CI -0.85 to -0.11; p = 0.011), and for MDD a trend of decreased IL-6 levels was observed (g: -0.39; CI -0.87 to 0.09; p = 0.115). Tumor necrosis factor alpha (TNFα) also decreased in schizophrenia (g: -0.34; CI -0.68 to -0.01; p = 0.047) and in MDD (g: -1.02; CI -1.79 to -0.25; p = 0.009). Fasting glucose levels increased only in schizophrenia (g: 0.26; CI 0.07-0.44; p = 0.007), but not in MDD. No changes were found for C-reactive protein, IL-1β, IL-2 and IL-4. Psychopharmacological treatment has modulating effects on BDNF and TNFα in drug-naïve first-episode patients with either schizophrenia or MDD. These findings support efforts for further research into transdiagnostic preventive strategies and augmentation therapy for those with immune dysfunctions.


Understanding the Relationship between Chronic Systemic Disease and Lichen Sclerosus Urethral Strictures.
Lichen sclerosus is a chronic, inflammatory skin condition of the genitalia of unknown origin that accounts for nearly 10% of urethral stricture disease. In this study we determine systemic comorbidities associated with lichen sclerosus in men. We analyzed data from 1,151 men who were enrolled in a multi-institutional prospective urethroplasty outcomes database. Individuals were grouped by stricture etiology, and baseline demographics, medical histories and patient reported outcome measures were retrospectively compared across groups. Of the 1,151 men in the database 81 (7.0%) were noted to have lichen sclerosus related urethral stricture disease. Average patient age was 46.06 ± 16.52 years, with those with lichen sclerosus being significantly older than those without lichen sclerosus (51.26 ± 13.84 vs 45.68 ± 16.64, p = 0.0011). Men with lichen sclerosus were more likely to have hypertension, hyperlipidemia and diabetes, and to use tobacco products. Controlling for age, men with lichen sclerosus related urethral stricture disease had a higher body mass index (aOR 1.089, 95% CI 1.050-1.130), and were more likely to have hypertension (aOR 2.028, 1.21-3.41) and be active tobacco users (aOR 2.0, 1.36-3.40). Mean preoperative patient reported outcome measures scores for urinary and sexual function were similar. Controlling for stricture length and location, the adjusted odds of surgical failure were higher for lichen sclerosus related urethral stricture disease (aOR 1.9, 95% CI 0.9-4.2). Lichen sclerosus related urethral stricture disease is associated with chronic systemic diseases. This association may implicate a systemic inflammatory and/or autoimmune pathophysiology. A 2-hit mechanism implicating local and systemic factors for lichen sclerosus related urethral stricture disease development and progression is hypothesized.


Use of a nutrition support protocol to increase enteral nutrition delivery in critically ill patients.
Early enteral nutrition is recommended for patients in intensive care units, but nutrition provision is often hindered by a variety of unit-specific problems. To evaluate the impact of a nutrition support protocol on nutrition prescription and delivery in the intensive care unit. Nutrition-related data from 73 patients receiving mechanical ventilation who were treated in an adult medical intensive care unit before introduction of an enteral nutrition support protocol were retrospectively compared with data for 87 patients admitted after implementation of the protocol. After implementation of the protocol, enteral nutrition was started significantly earlier (P = .007) and enteral feeding goals were reached significantly faster (6 vs 10 days, P < .001) than before. Prescription of enteral nutrition on the first day of invasive mechanical ventilation increased from 38% before to 54% after (P = .03) implementation of the protocol. Prescribed and delivered nutrition doses on the first 2 days of mechanical ventilation also increased significantly (P < .001) after the protocol was implemented. Nasojejunal feeding tubes were used in 52% of patients before and 56% of patients after protocol implementation P = .63). Jejunal tubes were placed earlier after the protocol was implemented than before (median 5 vs 6.5 days), and when a jejunal tube was in place, feeding goals were reached faster (median 2 vs 3 days, P = .002). Implementing an enteral nutrition support protocol shortened the time to reach feeding goals. Jejunal feeding tubes were necessary in more than half of the patients, and with a jejunal feeding tube in place, feeding goals were reached rapidly.


Diagnosis and treatment of autoimmune pancreatitis.
Clinical management of autoimmune pancreatitis changed over the last years. The lack of high-quality studies, probably due to the relative rarity of the disease, has not fully clarified many diagnostic and therapeutic aspects. Aim of this review is to overview the literature from a clinical point of view, focusing on diagnostic and therapeutic approach to this complicated disease. They have been focused mainly on the risk of misdiagnosing a cancer. Many significant articles have been published on the treatment strategies of autoimmune pancreatitis, considering both induction and maintenance therapy. Diagnosis of autoimmune pancreatitis remains challenging, particularly in focal pancreatic involvement, despite recent advances on imaging modalities. Treatment is based on induction and maintenance phases. Steroid treatment remains the best option to induce remission. Maintenance therapy may be used to prevent disease relapses, and low-dose steroids, azathioprine or rituximab are the therapeutic options. However, it remains unclear which patient needs to be treated.


Ion-pairing π-electronic systems: ordered arrangement and noncovalent interactions of negatively charged porphyrins.
In this study, charged π-electronic species are observed to develop stacking structures based on electrostatic and dispersion forces. i π- i π Interaction, defined herein, functions for the stacking structures consisting of charged π-electronic species and is in contrast to conventional π-π interaction, which mainly exhibits dispersion force, for electronically neutral π-electronic species. Establishing the concept of i π- i π interaction requires the evaluation of interionic interactions for π-electronic ion pairs. Free base (metal-free) and diamagnetic metal complexes of 5-hydroxy-10,15,20-tris(pentafluorophenyl)porphyrin were synthesized, producing π-electronic anions upon the deprotonation of the hydroxy unit. Coexisting cations in the ion pairs with porphyrin anions were introduced as the counter species of the hydroxy anion as a base for commercially available cations and as ion-exchanged species, via Na+ in the intermediate ion pairs, for synthesized π-electronic cations. Solid-state ion-pairing assemblies were constructed for the porphyrin anions in combination with aliphatic tetrabutylammonium (TBA+) and π-electronic 4,8,12-tripropyl-4,8,12-triazatriangulenium (TATA+) cations. The ordered arrangements of charged species, with the contributions of the charge-by-charge and charge-segregated modes, were observed according to the constituent charged building units. The energy decomposition analysis (EDA) of single-crystal packing structures revealed that electrostatic and dispersion forces are important factors in stabilizing the stacking of π-electronic ions. Furthermore, crystal-state absorption spectra of the ion pairs were correlated with the assembling modes. Transient absorption spectroscopy of the single crystals revealed the occurrence of photoinduced electron transfer from the π-electronic anion in the charge-segregated mode.


Expansion of the phenotype of Kosaki overgrowth syndrome.
Skeletal overgrowth is a characteristic of several genetic disorders that are linked to specific molecular signaling cascades. Recently, we established a novel overgrowth syndrome (Kosaki overgrowth syndrome, OMIM #616592) arising from a de novo mutation in PDGFRB, that is, c.1751C>G p.(Pro584Arg). Subsequently, other investigators provided in vitro molecular evidence that this specific mutation in the juxtamembrane domain of PDGFRB causes an overgrowth phenotype and is the first gain-of-function point mutation of PDGFRB to be reported in humans. Here, we report the identification of a mutation in PDGFRB, c.1696T>C p.(Trp566Arg), in two unrelated patients with skeletal overgrowth, further confirming the existence of PDGFRB-related overgrowth syndrome arising from mutations in the juxtamembrane domain of PDGFRB. A review of all four of these patients with an overgrowth phenotype and PDGFRB mutations revealed postnatal skeletal overgrowth, premature aging, cognitive impairment, neurodegeneration, and a prominent connective tissue component to this complex phenotype. From a functional standpoint, hypermorphic mutations in PDGFRB lead to Kosaki overgrowth syndrome, infantile myofibromatosis (OMIM #228550), and Penttinen syndrome (OMIM #601812), whereas hypomorphic mutations lead to idiopathic basal ganglia calcification (OMIM #615007). In conclusion, a specific class of mutations in PDGFRB causes a clinically recognizable syndromic form of skeletal overgrowth.


A review of the intraosseous course of the nerves of the mandible.
A review of the literature revealed that a common feature of all human mandibles is a large nerve (inferior alveolar nerve, IAN) between the mandibular and mental foramina. This nerve sends branches directly to the teeth or contributes a variable number of branches to a plexus of nerves which does the same. The plexus originates from a separate nerve that enters the mandibular foramen. The nerve plexus has been demonstrated by dissections and not by radiology. The buccal-lingual and superior-inferior positions of the IAN were not consistent among mandibles. The intramandibular IAN frequently ran a concave curve with a posterior segment descending as it progressed anteriorly and an anterior segment that ascended to the mental foramen. A bony canal was not always observed between the mandibular and mental foramina. The canal frequently lacked definite walls, especially near the mental foramen. Bilateral symmetry (location of the canal in each half of the mandible) was common, whereas duplications of the canal were rare. Nutrient canals and other branches of the mandibular nerve have been observed within the mandible. These may have been confused for the IAN or may contribute to the plexus of nerves.


Mithun (Bos frontalis): the neglected cattle species and their significance to ethnic communities in the Eastern Himalaya - A review.
This review consolidates the available information on the socio-economic and ecological significance of Mithun in the lives of ethnic communities in the Eastern Himalaya. Standard guidelines were followed for the review and data collection was carried out at three stages; literature search, literature screening, and literature review and analysis. Records indicate a long association of Mithun with the ethnic groups. Mithun serves as a symbol of pride and local currency for barter trade in the ethnic society. Its utilities range from being used as a bride price to settling legal disputes. Several cultural festivals and local ceremonies are celebrated around this bovine. Due to its semi-wild nature, this animal also has an ecological role to conserve broad leaf sub-tropical forests. However, it remains neglected and has not received policy attention, leading to a stagnated growth. The institutions for Mithun research and development are also weak. Furthermore, the species is under threat from new diseases and habitat alteration triggered by climate change. Founded on the current state of knowledge, there is a need for institutional development, strengthening institutional linkages, and promoting regional cooperation among Mithun rearing countries for further research and development of this unique cattle.


Prevalence of pre- and postpartum depression in Jamaican women.
Maternal depression during pregnancy has been studied less than depression in postpartum period. The aims of this study were to find out the prevalence of prepartum and postpartum depression and the risk factors associated in a cohort of Afro-Jamaican pregnant women in Jamaica. The Zung self-rating depression scale instrument was administered to 73 healthy pregnant women at 28 weeks gestation and at 6 weeks postpartum for quantitative measurement of depression. Blood samples were collected at 8, 28, 35 weeks gestation and at day 1 and 6 weeks postpartum to study the thyroid status. Study demonstrated depression prevalence rates of 56% and 34% during prepartum and postpartum period, respectively. 94% women suffering depression in both periods were single. There were significant variations in both FT3 and TT4 concentrations which increased from week 8 to week 28 prepartum (p < 0.05) and then declined at the 35th week (p < 0.05 compared with week 28) and 1 day post delivery study (p < 0.05 compared with week 35). The mean values for TSH increased significantly from week 8 through week 35. The mean values at 1 day postpartum and 6 week postpartum were not significantly different from the 35 week values. For FT3, TT4 and TSH there were no significant between group differences in concentrations. The major determinants of postpartum depression were moderate and severe prepartum depression and change in TT4 hormone concentrations. High prevalence of depression was found during pre- and postpartum periods. Single mothers, prepartum depression and changes in TT4 were factors found to be significantly associated with postpartum depression.


Whole mitochondrial genome sequence of a rat pancreatic adenocarcinoma CRL-2389 LTPA cell line.
Pancreatic adenocarcinoma is the fifth leading cause of cancer death in the world. We sequenced a complete mitochondrial genome sequence of a rat pancreatic tumor CRL-2389 LTPA cell line for the first time. The total length of the mitogenome was 16,314 bp and coding 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes. This mitochondrial genome sequence will provide new genetic resource into pancreatic tumor disease.


Using data from multiple studies to develop a child growth correlation matrix.
In many countries, the monitoring of child growth does not occur in a regular manner, and instead, we may have to rely on sporadic observations that are subject to substantial measurement error. In these countries, it can be difficult to identify patterns of poor growth, and faltering children may miss out on essential health interventions. The contribution of this paper is to provide a framework for pooling together multiple datasets, thus allowing us to overcome the issue of sparse data and provide improved estimates of growth. We use data from multiple longitudinal growth studies to construct a common correlation matrix that can be used in estimation and prediction of child growth. We propose a novel 2-stage approach: In stage 1, we construct a raw matrix via a set of univariate meta-analyses, and in stage 2, we smooth this raw matrix to obtain a more realistic correlation matrix. The methodology is illustrated using data from 16 child growth studies from the Bill and Melinda Gates Foundation's Healthy Birth Growth and Development knowledge integration project and identifies strong correlation for both height and weight between the ages of 4 and 12 years. We use a case study to provide an example of how this matrix can be used to help compute growth measures.


["Splint dishabituation" in treatment of distal injuries of the digital extensor tendons].
Results of treatment of 61 patients with "mallet finger" injuries have been analyzed. After discontinuation of permanent immobilization the patients of the main group (n = 20) underwent "splint dishabituation" consequentially and bit by bit during 8 weeks, according to an original protocol developed with special reference to the stages of biological process of union of the damaged tendon. Following permanent immobilization patients of the group of comparison (n = 35) were recommended to use a removable splint at night during two weeks. The proposed protocol of "splint dishabituation" improved the results of treatment of injuries of the "mallet finger" type.


Piperlongumine inhibits the proliferation and survival of B-cell acute lymphoblastic leukemia cell lines irrespective of glucocorticoid resistance.
Piperlongumine (PL), a pepper plant alkaloid from Piper longum, has anti-inflammatory and anti-cancer properties. PL selectively kills both solid and hematologic cancer cells, but not normal counterparts. Here we evaluated the effect of PL on the proliferation and survival of B-cell acute lymphoblastic leukemia (B-ALL), including glucocorticoid (GC)-resistant B-ALL. Regardless of GC-resistance, PL inhibited the proliferation of all B-ALL cell lines, but not normal B cells, in a dose- and time-dependent manner and induced apoptosis via elevation of ROS. Interestingly, PL did not sensitize most of B-ALL cell lines to dexamethasone (DEX). Only UoC-B1 exhibited a weak synergistic effect between PL and DEX. All B-ALL cell lines tested exhibited constitutive activation of multiple transcription factors (TFs), including AP-1, MYC, NF-κB, SP1, STAT1, STAT3, STAT6 and YY1. Treatment of the B-ALL cells with PL significantly downregulated these TFs and modulated their target genes. While activation of AURKB, BIRC5, E2F1, and MYB mRNA levels were significantly downregulated by PL, but SOX4 and XBP levels were increased by PL. Intriguingly, PL also increased the expression of p21 in B-ALL cells through a p53-independent mechanism. Given that these TFs and their target genes play critical roles in a variety of hematological malignancies, our findings provide a strong preclinical rationale for considering PL as a new therapeutic agent for the treatment of B-cell malignancies, including B-ALL and GC-resistant B-ALL.


Use of expenditure analysis to enhance returns on investments in HIV services.
Globally, the response to the HIV epidemic is at a crisis point. International investments in the HIV response have been essentially flat for 8 years and domestic budgets in low and middle-income countries - still recovering from the global recession - have not been able to fill the resource gap to drive a full-fledged HIV response. Still, efficiencies and prioritization of evidence-based interventions enable a significant scale-up of treatment, but millions more people remain without treatment. This review looks at recent data and research to evaluate interventions that may help close gaps in service provision that undermine testing and treatment programs. The President's Emergency Plan for AIDS Relief recently began publicly releasing vast programmatic and expenditure data. These data reveal potential efficiency gaps in testing and treatment programs, particularly in the area of linkage and retention. Interventions such as HIV self-testing have been proposed to help, but whether they can deliver better results remains unclear. Same-day initiation on treatment improves initiation, retention, and viral suppression rates. Near real-time analysis of data and active response is critical in improving efficiencies in programs. More investment in implementation research is necessary to improve linkage to care and treatment to reach 90-90-90 goals.


Carcinoma of the gallbladder: an appraisal of surgical resection.
After the development of hepatobiliary imaging modalities and surgical procedures, we reappraised the effect of surgical treatment on the prognosis of gallbladder carcinoma. Records of 32 patients with gallbladder carcinoma who underwent surgical resection between 1982 and 1992 were examined according to the pTNM staging system, and patients' prognoses were analyzed in relation to pathologic staging and surgical procedures. The standard operative procedure was extended cholecystectomy (cholecystectomy plus hepatic resection [wedge or anatomic segmentectomy] plus en bloc lymph node dissection) with or without extrahepatic bile duct resection, and this was performed in 63% of cases. The actual 5-year survival rate of patients whose postoperative period was more than 5 years was 53%. The standard operative procedure gave satisfactory results for nine patients with stage I or II disease because all except two patients who died of other diseases were still alive more than 3 years after operation and there were five (56%) 5-year survivors. At stage III with pN1a lymph node metastasis, 50% of patients survived more than 5 years. However, in patients with pN1b or distant organ metastases, none survived more than 3 years even after more aggressive operation including hepatectomy plus pancreatoduodenectomy. Complete removal of microscopic spread to the liver, bile duct, and lymph nodes is a great advantage to patients with gallbladder carcinoma at stages I and II and to those at stage III even in the presence of lymph node metastasis in the hepatoduodenal ligament.


[Creating the optimal microecology of the intestines in premature newborn infants with the sour-milk mixture "biphilin"].
The paper is concerned with formation of intestinal biocenosis in premature infants and with approaches to the correction of its disorders with the aid of biphilin, a special mixture prepared from sour milk. Ninety newborns showed the derangement of biocenosis of various intensity by the 6th to the 10th day of life. Application of biphilin for 2 to 6 weeks as part of the multimodality treatment of such children made the intestinal microflora return to normal.


Experimental sepsis: characteristics of activated macrophages and apoptotic cells in the rat spleen.
Sepsis, being characterized by massive translocation of bacteria into tissues, induces the suppression of the function of both leukocytes and macrophages. The aim of the study was to count activated macrophages (AMs) and apoptotic (Ao) cells in the rat spleen during the period of experimental sepsis and to clarify the associations of these parameters with each other and with leukocyte migration and bacterial translocation into different organs. The Wistar rats were intraperitoneally inoculated with Escherichia coli (E. coli) and were sacrificed after 2, 6, 24, 48, and 120 h. Bacteria and leukocytes in tissues were specifically stained. AMs were identified by immunohistological staining and Ao cells by the TUNEL assay. The high counts of E. coli at 6 h were strongly associated with a low level of the total counts of leukocytes, accompanied by the high translocation of microbes into tissues. In the spleen, lymphocytes, macrophages, and neutrophils with pyknotic nuclei were identified. The count of AMs was highest at 24 h after the inoculation with E. coli; at the same time the Ao cell count began to rise and achieved the highest level 24 h later. Our investigation indicates that the molecular peculiarities of macrophages and their responses to the inflammation process are tissue-specific. In the spleen the activation process involving hematopoietic cells and macrophages was remarkable at the late stage of sepsis, characterized by a high count of Ao cells.


The risk of COVID-19 death is much greater and age dependent with type I IFN autoantibodies.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection fatality rate (IFR) doubles with every 5 y of age from childhood onward. Circulating autoantibodies neutralizing IFN-α, IFN-ω, and/or IFN-β are found in ∼20% of deceased patients across age groups, and in ∼1% of individuals aged <70 y and in >4% of those >70 y old in the general population. With a sample of 1,261 unvaccinated deceased patients and 34,159 individuals of the general population sampled before the pandemic, we estimated both IFR and relative risk of death (RRD) across age groups for individuals carrying autoantibodies neutralizing type I IFNs, relative to noncarriers. The RRD associated with any combination of autoantibodies was higher in subjects under 70 y old. For autoantibodies neutralizing IFN-α2 or IFN-ω, the RRDs were 17.0 (95% CI: 11.7 to 24.7) and 5.8 (4.5 to 7.4) for individuals <70 y and ≥70 y old, respectively, whereas, for autoantibodies neutralizing both molecules, the RRDs were 188.3 (44.8 to 774.4) and 7.2 (5.0 to 10.3), respectively. In contrast, IFRs increased with age, ranging from 0.17% (0.12 to 0.31) for individuals <40 y old to 26.7% (20.3 to 35.2) for those ≥80 y old for autoantibodies neutralizing IFN-α2 or IFN-ω, and from 0.84% (0.31 to 8.28) to 40.5% (27.82 to 61.20) for autoantibodies neutralizing both. Autoantibodies against type I IFNs increase IFRs, and are associated with high RRDs, especially when neutralizing both IFN-α2 and IFN-ω. Remarkably, IFRs increase with age, whereas RRDs decrease with age. Autoimmunity to type I IFNs is a strong and common predictor of COVID-19 death.


Extended validation of the Relational Depth Frequency Scale: Retest reliability, divergent validity, criterion validity with psychotherapy satisfaction, and measurement invariance in UK- and US-stratified samples.
The Relational Depth Frequency Scale (RDFS) assesses moments of profound connection in psychotherapy, associated with therapeutic benefit. To date, the RDFS has not been tested for its retest reliability, divergent and criterion validity, and measurement invariance, nor has it been tested in stratified samples of psychotherapy patients. Two stratified online samples of United Kingdom (n = 514) and United States (n = 402) psychotherapy patients filled out the RDFS, the Brief Social Desirability Scale (BSDS); and the Satisfaction with Therapy and Therapist Scale-revised (STTS-R). Two subsamples of patients (United Kingdom: n = 50 and United States: n = 203) filled out the RDFS again after 1 month. Reliability for the six-item RDFS were excellent in United Kingdom and United States samples (Cronbach's α = 0.91 and 0.92; retest r = 0.73 and r = 0.76). Divergent (r = 0.10 and r = 0.12) and criterion validity (r = 0.69; and r = 0.70) were good. Full scalar invariance was established across countries, genders, and time. This contributes important evidence to the validity of the RDFS. Future research should assess predictive validity against psychotherapy outcomes and replicate these analyses in diverse samples.


Clinical features of scrupulosity: Associated symptoms and comorbidity.
Scrupulosity is a manifestation of obsessive-compulsive disorder (OCD) characterized by religious or moral core fears. Clinicians often struggle to treat scrupulosity, which may be associated with several features known to predict poor treatment outcome. The purpose of this study was to examine these features in participants with scrupulous OCD, contamination OCD, and healthy controls. A total of 68 participants (57.4% women, agemean = 34.01) completed diagnostic interviews, and measures of symptoms and quality-of-life. Relative to comparison groups, scrupulous participants had higher rates of obsessive-compulsive personality disorder, more severe schizotypal symptoms, and more severe symptoms of depression. In addition, OCD severity was strongly associated with poor insight in the scrupulous group. Both OCD groups reported poorer quality of life than did healthy controls. Clinicians working with scrupulous individuals may enhance the efficacy of treatment in this challenging population by assessing carefully for these features, and incorporating treatment elements that address them.


Learning to perform endoscopic resection of esophageal neoplasia is associated with significant complications even within a structured training program.
Endoscopic resection is the cornerstone of endoscopic treatment of esophageal high grade dysplasia or early cancer. Endoscopic resection is, however, a technically demanding procedure, which requires training and expertise. The aim of the current study was to prospectively evaluate efficacy and safety of the first 120 endoscopic resection procedures of early esophageal neoplasia performed by six endoscopists (20 endoscopic resections each) who were participating in an endoscopic resection training program. The program consisted of four tri-monthly 1-day courses with lectures, live-demonstrations, hands-on training on anesthetized pigs, and one-on-one hands-on training days. Gastroenterologists from centers with multidisciplinary expertise in upper gastrointestinal oncology participated, together with an endoscopy nurse and a pathologist. Outcome measures were complete endoscopic removal of the target area and acute complications. A total of 120 consecutive esophageal endoscopic resection procedures (85 ER-cap, 35 multiband mucosectomy [MBM]) were performed by six endoscopists: 109 in Barrett's esophagus, 11 for squamous neoplasia; 85 piecemeal endoscopic resections (median 3 specimens, interquartile range 2 - 4 specimens). Complete endoscopic removal was achieved in 111 /120 cases (92.5 %). Six perforations occurred (5.0 %): five were effectively treated endoscopically (clips, covered stent), and one patient underwent esophagectomy. There were 11 acute mild bleedings (9.2 %), which were managed endoscopically. Perforations occurred in ER-cap procedures performed by four participants (7.1 % ER-cap vs. 0 % MBM; P = 0.18), and in 1.7 % of the first 10 endoscopic resections and 8.3 % of the second 10 endoscopic resections per endoscopist (P = 0.26). In this intense, structured training program, the first 120 esophageal endoscopic resections performed by six participants were associated with a 5.0 % perforation rate. Although perforations were adequately managed, performing 20 endoscopic resections may not be sufficient to reach the peak of the learning curve in endoscopic resection.


Incidental learning of sound categories is impaired in developmental dyslexia.
Developmental dyslexia is commonly thought to arise from specific phonological impairments. However, recent evidence is consistent with the possibility that phonological impairments arise as symptoms of an underlying dysfunction of procedural learning. The nature of the link between impaired procedural learning and phonological dysfunction is unresolved. Motivated by the observation that speech processing involves the acquisition of procedural category knowledge, the present study investigates the possibility that procedural learning impairment may affect phonological processing by interfering with the typical course of phonetic category learning. The present study tests this hypothesis while controlling for linguistic experience and possible speech-specific deficits by comparing auditory category learning across artificial, nonlinguistic sounds among dyslexic adults and matched controls in a specialized first-person shooter videogame that has been shown to engage procedural learning. Nonspeech auditory category learning was assessed online via within-game measures and also with a post-training task involving overt categorization of familiar and novel sound exemplars. Each measure reveals that dyslexic participants do not acquire procedural category knowledge as effectively as age- and cognitive-ability matched controls. This difference cannot be explained by differences in perceptual acuity for the sounds. Moreover, poor nonspeech category learning is associated with slower phonological processing. Whereas phonological processing impairments have been emphasized as the cause of dyslexia, the current results suggest that impaired auditory category learning, general in nature and not specific to speech signals, could contribute to phonological deficits in dyslexia with subsequent negative effects on language acquisition and reading. Implications for the neuro-cognitive mechanisms of developmental dyslexia are discussed.


Analysis of antibody responses to predominant linear epitopes of Theiler's murine encephalomyelitis virus.
Using synthetic peptides, we have defined the major linear antibody epitopes of Theiler's murine encephalomyelitis virus (TMEV), i.e., A1A (VP1(12-25)), A1Ba (VP1(146-160)), A1Cb (VP1(262-276)), A2A (VP2(2-16)), A2B (VP2(165-179)), and A3A (VP3(24-37)). A time course study with either pooled or individual sera indicates that susceptible SJL mice intracerebrally infected with TMEV strongly and selectively recognize the A1Cb epitope of VP1, compared with resistant BALB/c or C57BL/6 mice, which broadly recognize most of the epitopes on the different capsid proteins. However, antibodies from SJL mice subcutaneously immunized with TMEV recognize primarily A1Ba, A1Cb, and A2A epitopes. A similar predominant recognition of the A1Cb epitope was found with antibodies from the cerebrospinal fluid of intracerebrally virus-infected SJL mice. Interestingly, a substantial level of antibodies against the A1Cb epitope in virus-infected SJL mice is of the immunoglobulin G2a subclass, in contrast to an undetectable level of this immunoglobulin G subclass in virus-immunized SJL mice. The level of in vitro viral neutralization by antibodies did not correlate with the clinical signs. Antibodies to A1Cb, A2A, and A2B were able to neutralize viral plaque formation in vitro, while antibodies to A3A, A1A, and A1Ba were not.


Social skills problems in neurotic outpatients. Social skills training with and without cognitive modification.
Twenty-two "socially dysfunctional" outpatients had 12 1 1/2-hour sessions of social skills training either alone or combined with cognitive modification (each, n = 11). Two raters who were blind to the treatment regimen significantly differentiated the two treatments, as judged from audiotapes of treatment sessions. Measurement was by an independent assessor in a structured interview, by self-monitoring of several activities and of anxiety felt during them, and by self-rated questionnaires. In each treatment regimen, patients' conditions improved significantly and equally on reported behavioral and cognitive measures both during and after treatment, and at six-month follow-up. Outcome did not differ significantly between the two treatment conditions at any time on any measure. Patients reported increased levels of social activities and less anxiety during these, less isolation, better relations with their colleagues at work, less depression, and loss of many irrational social beliefs. During multiple baseline monitoring for up to nine weeks, no improvement occurred. Performance of each social task or "target" increased significantly after training was begun for it, and subjective anxiety decreased similarly during social performance. Untreated targets improved much less, and more slowly. Social skills training was followed by lasting and worthwhile improvement that was not enhanced by the addition of cognitive modification.


Use of antifungal therapy in hospitalized patients. II. Results after the marketing of fluconazole.
To evaluate the prescribing patterns of antifungal agents in the hospital setting after the introduction of fluconazole, a new broad-spectrum bis-triazole antifungal agent. Also compared are the prescribing patterns of antifungal agents prior to (phase I) and following (phase II) fluconazole marketing. A prospective cohort of hospitalized patients prescribed topical or systemic antifungal agents. Data were collected from December 1990 to April 1991. Fifty-seven hospitals ranging in size from 100 to more than 500 beds. Sixty-three percent are affiliated with medical schools. Participating pharmacists consecutively identified 15 patients receiving systemic antifungal therapy and 5 patients receiving topical antifungal therapy. Observational data on patient antifungal therapy, risk factors for fungal infections, comorbidities, concurrent medications, and culture data were collected. Differences in prescribing patterns before and after the marketing of fluconazole were assessed using t-tests and chi-square tests. Of 818 patients studied, 615 (75.2 percent) received systemic antifungal therapy. Five hundred forty-six patients received a single antifungal agent; 348 (63.7 percent) received fluconazole, 105 (19.2 percent) received ketoconazole, 92 (16.8 percent) received amphotericin B, and 1 (0.2 percent) received flucytosine. Sixty-nine patients received two or more systemic agents either concurrently or consecutively. The use of parenteral amphotericin B, alone or in combination with flucytosine and/or an azole, declined from 56.8 percent in the phase I study to 24.2 percent in the current study. The use of parenteral therapy also declined from 56.8 to 40.2 percent. Ketoconazole was used in more than 90 percent of the oral and esophageal infections in the phase I study, but its use declined to only 33 percent in this study. Fluconazole was used most frequently across all sites of presumed or documented infections, with the exception of fungemia. Of the presumed or proven systemic or blood infections, amphotericin B was used alone or in combination in 48.4 percent of the patients and fluconazole was used exclusively in 39.0 percent of the patients. Fluconazole was used more often than amphotericin B (22 vs. 3 patients, respectively) for prophylaxis of systemic infections. The overall use of antifungal prophylaxis also increased from the phase I (9.5 percent) to phase II (13.7 percent). The introduction of fluconazole had a major impact on the prescribing patterns of antifungal therapy. Although amphotericin B remained the preferred agent for treatment of suspected or proven systemic, central nervous system, or blood infections, use of fluconazole for these indications approached nearly 40 percent. Further studies are needed to address the role of fluconazole in the prophylaxis and treatment of systemic mycoses.


Walk with a pen, or fly with a computer. Can electronic dentistry work for you?
In the last several years, dentistry has crossed over into the new frontier of electronic dentistry. It has embraced such developments as computer programs for producing digital radiographs and photographs, as well as digital programs that enhance these images, store and organize them into a retrievable "chart-like" fashion, and transmit them via the Internet. In Europe, I saw patients with an electronic "health card." This credit card-sized CD can carry all the information on a patient's written charts, results of laboratory tests, radiographic/imaging information and more. It is expected that the mobile phone will be an alternate vehicle for patient records, and that these records will be accessed with a password security system. This will allow patients to carry their records from location to location. Certainly, the dental implications of such seemingly advanced processes are evident. The expression, "The future is now," was never truer.


TGF-beta and CD4+CD25+ regulatory T cells.
TGF-beta and CD4+CD25+ regulatory T cells (Treg) both play an important role in the control of immune responses and the maintenance of immune homeostasis. The mechanism of suppression induced by Treg and the factors which regulate Treg function and number, have only begun to be elucidated. TGF-beta seems to act as an effector cytokine involved in the immunosuppressive function of Treg in vitro and in vivo, although its origin and mechanism of action remains to be defined. In addition, TGF-beta signaling in peripheral Treg seems to be essential for the regulation of peripheral Treg numbers and for their immunosuppressive function in vivo. This review will focus on the role of TGF-beta for the generation and expansion of CD4+CD25+ Treg, as well as for their immunosuppressive function in vitro and in vivo.


Distribution patterns of apolipoproteins A1, A2, and B in the wall of atherosclerotic vessels.
Atherosclerotic vessels were analysed histochemically for distribution, quantity, and composition of apolipoprotein (Apo) types in the vascular wall. The specimens comprised all stages of atherosclerosis, from very discrete intimal changes to complicated lesions. The vessel specimens were marked with antibodies against human Apo A1, A2, and B. Apo A1 can be demonstrated in even the earliest stage of atherosclerosis, and increases with the progression of the disease. In the initial stage, Apo A1 is found first in lumen-adjacent layers of the intima, and is evident in deeper layers of the wall as the disease progresses. Arteries of muscular type show accumulation of Apo in an earlier stage (or in greater quantity at the same stage) than arteries of elastic type. At all stages, the amount of Apo A1 always exceeds that of A2 and B. In the intima, Apo B is higher than Apo A2, the media contains hardly any Apo B, and the adventitia has less B than A2. Within the intimal layer, Apo A1 and A2 are found in an intracellular (mainly in foam cells) or in an extracellular location, according to the stage of atherosclerosis. Apo B is almost exclusively extracellular; only cases of advanced atherosclerosis show some intracellular localization (mostly in foam cells), visualized as electron dense lamellar organelles, probably of lysosomal origin. In the media, Apo A1 and A2 are accumulated in intracellular deposits, whereas the extracellular storage of Apo A1, A2 and B is observed only in cases with the most severe damage. Our investigations suggest that the accumulation of apolipoproteins in the vascular wall is affected not only by insudation from the plasma, but also by neosynthesis and/or metabolism by locally derived cells or cells immigrating in the process of atherosclerosis. The presence of Apo A1 and A2 in the vessel wall is now documented, and their role at this site apparently differs from that in the plasma.