Two distinct insulin-related molecules in the guinea pig: immunological and biochemical characterization of insulin-like immunoactivity from extrapancreatic tissues of the guinea pig.
In this study we extracted guinea pig brain and testis with; the extract was adsorbed to and eluted from cartridges (the Sep-Pak C18 procedure). We found this procedure superior for recovering crystalline insulin added to buffers or tissues, and for recovering endogenous insulin from plasma, but inferior for recovery of insulin from tissues. However, we did find 'rat/pork' type-insulin in guinea pig brain and testis (5-50 pg/g wet weight tissue). Our results with the Sep-Pak C18 procedures were reproduced by four other laboratories (who found 4-60 pg/g wet weight of tissue) and similar findings were also obtained by an independent investigator. Thus, we conclude that extrapancreatic tissues of guinea pigs have a second type of insulin-related material that is more typical of other mammalian insulins, but that the amount recovered is dependent upon the extraction procedure utilized.


Providing supportive care to cancer patients: a study on inter-organizational relationships.
Supportive cancer care (SCC) has historically been provided by organizations that work independently and possess limited inter-organizational coordination. Despite the recognition that SCC services must be better coordinated, little research has been done to examine inter-organizational relationships that would enable this goal. The purpose of this study was to describe relationships among programs that support those affected by cancer. Through this description the study objective was to identify the optimal approach to coordinating SCC in the community. Senior administrators in programs that provided care to persons and their families living with or affected by cancer participated in a personal interview. South-central Ontario, Canada. administrators from 43 (97%) eligible programs consented to participate in the study. Network analysis revealed a diffuse system where centralization was greater in operational than administrative activities. A greater number of provider cliques were present at the operational level than the administrative level. Respondents identified several priorities to improve the coordination of cancer care in the community including: improving standards of care; establishing a regional coordinating body; increasing resources; and improving communication between programs. Our results point to the importance of developing a better understanding on the types of relationships that exist among service programs if effective integrated models of care are to be developed.


Comparison of conventional stool concentration and preserved-smear methods with Merifluor Cryptosporidium/Giardia Direct Immunofluorescence Assay and ProSpecT Giardia EZ Microplate Assay for detection of Giardia lamblia.
Giardiasis is the most common parasitic infection in the United States. Variation in the numbers of cysts and/or trophozoites that are present along with the need for a skilled microscopist offer challenges in diagnosis. We compared the sediment wet preparation and permanent stained smear results (concentration in formalin-ethyl acetate and preparation of a smear prepared from a polyvinyl alcohol-preserved specimen) from 512 consecutive specimens with the results obtained by using the Merifluor Cryptosporidium/Giardia Direct Immunofluorescence Assay (DFA; Meridian Diagnostics, Inc., Cincinnati, Ohio) and the ProSpecT Giardia EZ Microplate Assay (EIA; Alexon, Inc., Sunnyvale, Calif.). The Merifluor DFA detected 33 of 33 positive specimens, and the ProSpecT EIA detected 32 of 33 positive specimens. The diagnostic sensitivities of the Merifluor DFA and the ProSpecT EIA were 100 and 97%, respectively. The specificities of the assays were 99.8%. The Merifluor DFA and the ProSpecT EIA appear to be equally sensitive, and both are more sensitive than conventional microscopy.


Injury patterns for occupants of small trucks.
A limited number of studies have been completed on the factors contributing to accident-related injuries sustained by occupants of pickup trucks. The increasing number and changing pattern of use of light trucks necessitates the need to critically review this vehicle type with respect to contributing accident factors and associated injuries. This paper investigates the injury mechanisms of occupants of pickup trucks and the surfaces that the occupants contact in roadway accidents. Selection of cases from the Canadian vehicle database was based on the location of the vehicles' most severe impact deformation. The overall occupant injury severity was examined with respect to impact location, impact speed, and magnitude of occupant compartment intrusion. The results of the investigation illustrate the benefits of seat belt restraint use. In addition, identification of frequent injury contact surfaces establishes a reference for improved vehicle design initiatives and standards.


'Space to talk': a Portuguese focus group study of parents' experiences, needs and preferences in parenting support during prenatal and well-child care.
To explore the experiences, needs and preferences of a group of parents regarding the parenting support received during prenatal and well-child care in the Portuguese National Health Service. We undertook descriptive-interpretive qualitative research running multiple focus groups in Porto, Northern Portugal. Purposive sampling was used between April and November 2018. Focus groups were conducted with 11 parents of a 0-3 years old with well-child visits done in primary care units. Thematic analysis was performed in a broadly inductive coding strategy and findings are reported in accordance with Consolidated Criteria for Reporting Qualitative Research guidelines. Three main themes were identified to describe parents' experience when participating in their children's healthcare: (1) logistics/delivery matter, including accessibility, organisation and provision of healthcare activities, unit setting and available equipment; (2) prenatal and well-child care: a relational place to communicate, with parents valuing a tripartite space for the baby, the family and the parent himself, where an available and caring health provider plays a major role and (3) parenting is challenging and looks for support, based on key points for providers to watch for and ask about, carefully explained and consensual among health providers. This study provides insight into parents' needs and healthcare practices that affect the parenting experience. To meet parents' preferences, sensitive health providers should guarantee a relational place to communicate and person-centredness, accounting for the whole family system to support healthy parenting collaboratively. Future studies are warranted to further strengthen the knowledge in the field of a population-based approach for parenting support.


Voice Quality as Therapeutic Outcome in Laryngopharyngeal Reflux Disease: A Prospective Cohort Study.
To study the usefulness of voice quality as therapeutic outcome in laryngopharyngeal reflux disease. A total of 80 patients with reflux finding score (RFS) > 7 and reflux symptom index (RSI) > 13 were treated with pantoprazole, diet, and lifestyle recommendations for 3 months. The therapeutic effectiveness was assessed with RSI; RFS; Voice Handicap Index; blinded Grade, Roughness, Breathiness, Asthenia, Strain, and Instability (GRBASI); aerodynamic and a panel of acoustic measurements before and after treatment. A correlation analysis between symptoms, videolaryngostroboscopic signs, and acoustic measurements was conducted. Compared to baseline, means of RSI, RFS, Voice Handicap Index, perceptual dysphonia, and roughness significantly decreased. Significant improvements of phonatory quotient, percent jitter, percent shimmer, Relative Average Perturbation, Pitch Perturbation Quotient, Phonatory F0 Range, Amplitude Perturbation Quotient, smooth Amplitude Perturbation Quotient, and Peak-to-Peak Amplitude Variation were found at the end of treatment. Studies of correlation did not identify relevant correlation between videolaryngostroboscopic signs, especially vocal folds edema, and objective voice quality evaluations. Voice quality assessments can help to better understand voice disorders and can be used as indicators of the treatment effectiveness in patients with laryngopharyngeal reflux-related symptoms.


The mechanism of immune dysregulation caused by porcine reproductive and respiratory syndrome virus (PRRSV).
PRRSV is capable of evading the effective immune response, thus persisting in piglets and throughout the swine herd. We show here that PRRSV invades the thymus and causes depletion of T-cell precursors and alteration of the TCR repertoire. Developing thymocytes are affected during negative selection when they transit from the triple-negative to triple-positive stages at the corticomedullary junction just before entering the medulla. The restriction of repertoire diversification occurs in both helper and cytotoxic αβ-T cells. As a result, critical viral epitopes are tolerated, and infection becomes chronic. However, not all viral epitopes are tolerated. Infected piglets develop antibodies capable of recognizing PRRSV, but these are not virus neutralizing. Further analysis showed that the lack of an effective immune response against the critical viral structures results in the absence of a germinal center response, overactivation of T and B cells in the periphery, robust production of useless antibodies of all isotypes, and the inability to eliminate the virus. Overall, the results show how a respiratory virus that primarily infects and destroys myelomonocytic cells has evolved strategies to disrupt the immune system. These mechanisms may be a prototype for how other viruses can similarly modulate the host immune system.


Solvent effects on fluorescence properties of protochlorophyll and its derivatives with various porphyrin side chains.
Fluorescence spectra and fluorescence lifetimes of protochlorophyll (Pchl) were measured in organic solvents having different physical and chemical properties and were analyzed taking into account the nonspecific (dependent on bulk solvent parameters), and specific (e.g. H bonds, Mg coordination) solvent-solute interactions. The energy of the fluorescence emission band decreased, while the Stokes shift increased for increasing solvent orientation polarizability, which is a function of both the dielectric constant (epsilon) and the refractive index (n). The extent of the dependence of the Stokes shift on solvent orientation polarizability was higher in protic (i.e. those able to form hydrogen-binding) than in aprotic solvents. High value of the Stokes shift was also observed in pyridine and methanol, i.e. in solvents hexacoordinating the central Mg atom. The fluorescence decay of Pchl was monoexponential in all of the investigated solvents. The fluorescence lifetime decreased for increasing solvent orientation polarizability from 5.5 +/- 0.1 ns in 1,4-dioxane to 3.3 +/- 0.1 ns in methanol. Longer lifetime values were observed in the case of aprotic solvents than in protic solvents. The hexacoordination of Mg had no effect on the fluorescence lifetime. The present data are discussed with respect to results found for protochlorophyllide (Pchlide) (Myśliwa-Kurdziel et al. in Photochem Photobiol 79:62-67, 2004), and they indicate that the presence of phytol chain in the porphyrin ring influences the spectral properties of the whole chromophore. This is the first complex analysis comparing the fluorescence emission and fluorescence lifetimes of purified Pchl and Pchlide.


Use of larval weight and length as indices of gut content weight in some lepidopterous larvae.
Significant correlation was obtained between body weight or length and the weight of contents of gut in chosen lepidopterous larvae ranging from 50 to 3,300 mg or 13 to 75 mm. The obtained relations can be used in the determination of net weight of a larva (i.e. the weight of the larva excluding its gut contents) and hence may find wide application in ecological studies.


The fluorescence amplification strategy based on 3D DNA walker and CRISPR/Cas12a for the rapid detection of BRAF V600E.
Circulating tumor DNA (ctDNA) is a fragment of single- or double-stranded DNA originating from tumor or circulating tumor cells and provides accurate information regarding the molecular characteristics of tumors. Therefore, sensitive detection of ctDNA is of great significance to mutation analysis and clinical diagnosis. Among various ctDNAs, the BRAF V600E is related to aggressive behavior, disease recurrence, and disease-specific mortality in papillary thyroid carcinoma. Herein, we selected the BRAF V600E gene sequence as an in vitro biomarker, and established a fluorescence detection strategy combined 3D DNA walker with CRISPR/Cas12a. In the presence of the target ctDNA, 3D DNA walker could identify and bind it, and thus released a large amount of output DNAs through cyclic cleavage with the assistance of specific endonuclease (Nb.BbvCI). The output DNAs were specifically bound to crRNA and activated the non-specific trans-cleavage activity of Cas12a. Finally, the fluorescence signal was significantly enhanced. Notably, this method can detect the BRAF V600E in a range of 1 fM ~ 20 nM with a detection limit of 0.37 fM without DNA polymerase. Due to the powerful amplification capability of 3D DNA walker and high specificity and programmability of CRISPR/Cas12a, the entire process took only a maximum of 70 min. Furthermore, it can be potentially used for the detection of ctDNA in human serum. In summary, this method not only provides a platform for the rapid detection of ctDNA, but also shows good potential for early clinical diagnosis and biomedical research.


Effect of Schistosoma mansoni infection, starvation and molluscicides on acid phosphate, transaminases and total protein in tissues and hemolymph of Biomphalaria alexandrina.
The activities of GOT and GPT in the hemolymph of B. alexandrina were significantly decreased by S. mansoni infection. However, the total protein concentration and AcP activity were increased. Although the snail starvation decreased AcP activity in the ovotestis, it increased GOT activity in the other organs of the snails. On the other hand, the snail feeding after starvation increased significantly AcP activity in ovotestis. Natural and synthetic molluscicides inhibited the activities of GOT and GPT, however, they increased the total protein concentrations and AcP activities in the examined organs.


RSM optimization studies for cadmium ions adsorption onto pristine and acid-modified kaolinite clay.
Clay has been reported as an active absorbent for the removal of toxic heavy metals from aqueous medium. In this study, pristine and acid modified kaolinite clays (PKC and AMKC) were prepared, characterized using various analyses, and tested for Cd2+ ion adsorption from textile industry wastewater. After acid modification, the specific surface area of clay increased from 84.2 to 389.4 m2/g. Adsorption isotherm, kinetics and thermodynamics behaviour process were examined. The pH at (pHpzc) of 8.5 indicate that AMKC surface is positively charged for pH below the pHpzc attracting anions. Response surface methodology was used to investigate the effect of adsorption process factors on Cd2+ ion removal uptake. At the optimum process conditions of 45.3 °C temperature, 0.63 g/L adsorbent dosage, and 120.9 min contact time, the percentages of Cd2+ adsorbed by PKC and AMKC were 77.82% and 99.19%, respectively. Various models were employed to analyzed the kinetic and equilibrium data. The Pseudo-first order, Pseudo-second order and Intra-particle diffusion were used to evaluate the kinetic data, while the Langmuir, Freundlich and Temkin isotherm models were applied to analyzed the equilibrium data. The sorption kinetics was found to be best described by Pseudo-second order considering the high correlation coefficient (R2), smaller Chi-square (ᵪ2) and sum of square error (SSE). The Freundlich model was the most accurate in describing the equilibrium data followed by Langmuir and Temkin respectively. The thermodynamic reveal that the reaction is spontaneous and endothermic in nature, and increase in randomness between the adsorbent and adsorbate. The obtained activation energy (Ea) value suggest that the adsorption mechanism of Cd(II) is a physisorption dominated.


Detection of endogenous epidermal growth factor-like activity in the developing chick embryo.
Epidermal growth factor-like activity has been detected by radioreceptor assay and radioimmunoassay in the developing chick embryo. Very little activity could be detected prior to Day 8 of embryonic life (hatching is at Day 21). A peak of EGF activity was detectable by both assays over Days 10 to 12. The EGF activity then fell to virtually undetectable levels during Days 14 to 17. A later rise in RRA detectable EGF like activity was then observed over Days 18-20. The EGF activity from a Day 11 embryo chromatographed on high-performance liquid chromatography as a single peak, with very high recovery of activity, at a later elution position than mouse EGF or human EGF.


Quantification of sugars and sugar phosphates in Arabidopsis thaliana tissues using porous graphitic carbon liquid chromatography-electrospray ionization mass spectrometry.
This work reports the development and optimisation of a negative ion mode on-line LC-ESI-MS/MS method for the sensitive targeted analysis of the key glycolytic intermediates, sugars and sugar phosphates from plants, using a porous graphitic carbon (PGC) stationary phase and an MS compatible mobile phase. Using this newly developed method, separation and detection of a solution of standard compounds is achieved in less than 20min. Target metabolite compounds were identified in plant extracts from their characteristic retention times, and product ion spectra. This on-line PGC-ESI-MS/MS method shows good linearity over the concentration range 0-100microM, selectivity, short analysis time, and limits of detection of 0.1microM for disaccharides trehalose (Tre), sucrose (Suc), and maltose, and 1.5microM for hexose phosphates fructose-6-phosphate (Fru6P), glucose-1-phosphate (Glc1P), and glucose-6-phosphate (Glc6P), and phosphoenolpyruvate (PEP). This paper describes details of our method and its application to the simultaneous quantitative analysis of soluble sugars and sugar phosphates from Arabidopsis thaliana tissues. We have demonstrated the utility of our method for the analysis of biological samples by applying it to the simultaneous quantitation of changes in soluble sugars and sugar phosphates in A. thaliana Columbia-0 (Col-0) and its starchless phosphoglucomutase (pgm) mutant over a 12-h light/12-h dark growth cycle.


Clinical and microbiological outcomes in haematogenous spondylodiscitis treated conservatively.
Spondylodiscitis refers to infections of the intervertebral disc and the adjacent vertebral body. Although it is still considered a rare condition, its rate is projected to increase. Mortality rate is considered to be low, but an estimated one third of the survivors experience residual disabilities. Literature shows that uncomplicated spondylodiscitis can be adequately treated by early antibiotic therapy and immobilization. The aim of the study is to evaluate the outcome of conservative treatment in patients with haematogenous spondylodiscitis. All patients with haematogenous spondylodiscitis observed in two orthopaedic centres were retrospectively considered. The medical records, radiologic imaging, bacteriology results, treatment, and complications of all patients were reviewed. Thirty patients (median age 64 years, range 15-77, females 56.7%) were considered in the study, eight (26.7%) showed residual back pain at median follow-up of 117 weeks (range 104-189). A significant difference in SF-36 physical (P < 0.001), SF-36 mental function (P < 0.002), and Oswestry Disability Index (ODI) (P < 0.001) scores was observed among patients with residual local pain compared to the ones who had not. Methicillin-resistant Staphylococcus aureus (MRSA) infection and symptoms duration before the diagnosis were associated with an increased risk of persistent back pain and permanent disability. The most important negative determinants of SF-36 mental function were the age of patients (ρ = 0.36, P < 0.05), the duration of symptoms before the diagnosis (ρ = 0.44, P < 0.05) and MRSA infection (P = 0.006). Spondylodiscitis sustained by MRSA and the duration of symptoms before the diagnosis influenced negatively the physical status (P = 0.002) and ODI (ρ = 0.36, P < 0.05), respectively. Conservative approaches are safe and effective for patients without complications. A delayed diagnosis and MRSA infections are related to poor clinical outcome among patients treated by conservative treatment; this must be carried out scrupulously with close patient monitoring.


The winner takes it all: Competition drives clonal selection in gliomagenesis.
The exact mechanisms that govern clonal dynamics and selection during early tumorigenesis remain largely elusive. Ceresa et al. provide experimental and mathematical evidence that MYC-dependent competition between individual clones is one driving force of brain tumor evolution, adding a winner/loser aspect to the picture that complements existing concepts.


Development and characterization of a xylose-inducible gene expression system for Clostridium perfringens.
A xylose-inducible gene expression vector for Clostridium perfringens was developed. Plasmid pXCH contains a chromosomal region from Clostridium difficile (xylR-P(xy)(lB)): xylR, encoding the xylose repressor, xylO, the xyl operator sequence, and P(xylB), the divergent promoter upstream of xylBA encoding xylulo kinase and xylose isomerase. pXCH allows tightly regulated expression of the chloramphenicol acetyltransferase reporter and the α-toxin genes in response to the inducer concentration. Thus, pXCH could constitute a new valuable genetic tool for study of C. perfringens.


Distinct diarrhea profiles during outpatient chemotherapy.
Chemotherapy-induced diarrhea (CID) is a common symptom that occurs in 50 to 80% of patients. Given that the majority of the data on the occurrence and severity of CID is based on physician-rated toxicity criteria, this study's purposes were to identify subgroups of patients with distinct CID profiles and determine how these subgroups differ in terms of demographic and clinical characteristics; severity, frequency, and distress of CID; the co-occurrence of common GI symptoms; and QOL. Patients (n = 1133) completed the Memorial Symptom Assessment Scale six times over two cycles of chemotherapy. Latent profile analysis was used to identify subgroups of patients with distinct diarrhea profiles. Differences among these subgroups were evaluated using parametric and nonparametric statistics. Four distinct diarrhea profiles were identified: none (58.3%), decreasing (22.0%), increasing (5.2%), and high (14.5%). Compared with the none class, patients in the high class had a lower functional status, a worse comorbidity profile, were more likely to have gastrointestinal cancer, and were more likely to receive chemotherapy on a 14-day cycle. No differences were found among the classes in the percentages of patients who received chemotherapy with a targeted therapy. Given that CID occurred in over 40% of the patients, clinicians should assess for this symptom and other common GI symptoms and initiate appropriate pharmacologic and dietary interventions.


Selective autocatalytic reduction of NO from sintering flue gas by the hot sintered ore in the presence of NH3.
In this paper, the selective autocatalytic reduction of NO by NH3 combined with multi-metal oxides in the hot sintered ore was studied, and the catalytic activity of the hot sintered ore was investigated as a function of temperature, NH3/NO ratio, O2 content, H2O and SO2. The experimental results indicated that the hot sintered ore, when combined with NH3, had a maximum denitration efficiency of 37.67% at 450 °C, 3000 h(-1) gas hourly space velocity (GHSV) and a NH3/NO ratio of 0.4/1. Additionally, it was found that O2 played an important role in removing NOx. However, high O2 content had a negative effect on NO reduction. H2O was found to promote the denitration efficiency in the absence of SO2, while SO2 inhibited the catalytic activity of the sintered ore. In the presence of H2O and SO2, the catalytic activity of the sintered ore was dramatically suppressed.


Efficient infection of monkey cells with SV40 DNA. II. Use of low-molecular-weight DEAE-dextran for large-scale experiments.
With standard protocols for DNA infection, only a small percentage (about 4%) of monkey ells exposed to purified DNA of simian virus 40 (SV40) displays cytopathic effect or expresses viral T antigen. We have recently reported (Sompayrac and Danna, 1981) that by extending the time of exposure of BSC-1 cells to DNA in the presence of low concentrations of diethylaminoethyl (DEAE)-dextran, we can infect up to 50% of the cells. Because our protocol was devised for DEAE-dextran of mol. wt. 2 X 10(6), which is no longer commercially available, we have tested a low-molecular-weight polymer (mol. wt. 0.5 X 10(6) that can be purchased. Use of this reagent in our protocol resulted in marginally higher levels of infection than we found with the high-molecular-weight polymer. However, slightly more DNA (about 1.5 times as much) was needed to achieve saturation. With both reagents, the percentage of cells infected was proportional to time of exposure. Therefore, low-molecular-weight DEAE-dextran should be useful for large-scale DNA infections.


Immunoassay and antibody microarray analysis of the HUPO Plasma Proteome Project reference specimens: systematic variation between sample types and calibration of mass spectrometry data.
Four different immunoassay and antibody microarray methods performed at four different sites were used to measure the levels of a broad range of proteins (N = 323 assays; 39, 88, 168, and 28 assays at the respective sites; 237 unique analytes) in the human serum and plasma reference specimens distributed by the Plasma Proteome Project (PPP) of the HUPO. The methods provided a means to (1) assess the level of systematic variation in protein abundances associated with blood preparation methods (serum, citrate-anticoagulated-plasma, EDTA-anticoagulated-plasma, or heparin-anticoagulated-plasma) and (2) evaluate the dependence on concentration of MS-based protein identifications from data sets using the HUPO specimens. Some proteins, particularly cytokines, had highly variable concentrations between the different sample preparations, suggesting specific effects of certain anticoagulants on the stability or availability of these proteins. The linkage of antibody-based measurements from 66 different analytes with the combined MS/MS data from 18 different laboratories showed that protein detection and the quality of MS data increased with analyte concentration. The conclusions from these initial analyses are that the optimal blood preparation method is variable between analytes and that the discovery of blood proteins by MS can be extended to concentrations below the ng/mL range under certain circumstances. Continued developments in antibody-based methods will further advance the scientific goals of the PPP.


Molecular basis of double-stranded RNA-protein interactions: structure of a dsRNA-binding domain complexed with dsRNA.
Protein interactions with double-stranded RNA (dsRNA) are critical for many cell processes; however, in contrast to protein-dsDNA interactions, surprisingly little is known about the molecular basis of protein-dsRNA interactions. A large and diverse class of proteins that bind dsRNA do so by utilizing an approximately 70 amino acid motif referred to as the dsRNA-binding domain (dsRBD). We have determined a 1.9 A resolution crystal structure of the second dsRBD of Xenopus laevis RNA-binding protein A complexed with dsRNA. The structure shows that the protein spans 16 bp of dsRNA, interacting with two successive minor grooves and across the intervening major groove on one face of a primarily A-form RNA helix. The nature of these interactions explains dsRBD specificity for dsRNA (over ssRNA or dsDNA) and the apparent lack of sequence specificity. Interestingly, the dsRBD fold resembles a portion of the conserved core structure of a family of polynucleotidyl transferases that includes RuvC, MuA transposase, retroviral integrase and RNase H. Structural comparisons of the dsRBD-dsRNA complex and models proposed for polynucleotidyl transferase-nucleic acid complexes suggest that similarities in nucleic acid binding also exist between these families of proteins.


Induction of proliferation in resting B-cells by a factor released by activated mouse spleen cells.
Mouse spleen cells activated in a mixed lymphocyte reaction release a soluble factor, which induces a significant proliferative response in fresh mouse spleen cells. This proliferation inducing factor (PIF) was found to be heat stable (90 degrees C for 45 min) and also resistant to trypsin or chymotrypsin treatment. By using a sizing HPLC column, the molecular weight of PIF appears to be 25 kDa. Mouse spleen cells treated with anti-thy-1 + complement lost Con-A induced proliferative responses but responded well to PIF. B cell depleted spleen cells obtained by negative selection panning, did not respond to PIF. These results indicate that B cells proliferated in response to PIF. Polymixin-B, which blocks the B cell proliferative response to LPS, did not inhibit PIF induced proliferation.


Analytical applications of the electrochemiluminescence of tris(2,2'-bipyridyl)ruthenium(II) coupled to capillary/microchip electrophoresis: a review.
A comprehensive review on the development of analytical methods, by coupling electrochemiluminescence (ECL) detection with capillary electrophoresis (CE) and microchip electrophoresis (ME), is presented. After the description of the basic mechanism of ECL, the addition mode of luminescence reagent in CE-ECL system has been discussed. The analytical applications of the CE-ECL technique in terms of different analytes are also given. Due to the importance of ME as a separation method for the present and future, the ME detection methods based on ECL are considered in a relatively detailed way. Finally, possible trends for CE/ME-ECL in the near future are discussed.


Changes in Phenolic Compounds Profiles in Tea Extracts and the Composition of these Phenolic Compounds in Yogurt.
Green, white, and black tea water extracts are rich in phenolic compounds. The changes in phenolic compound profiles of green, white, and black tea (GT, WT, & BT respectively) water extracts and their respective yogurt were investigated. Three types of yogurt with tea water extracts were prepared, and the phenolic compound profiles were analyzed using the liquid chromatography-mass spectrometry (LC-MS) method. The present data found that flavonol glycosides such as kaempferol-3-rutinoside and quercetin-rhamnosylgalactoside or rutinoside were present in WT extract, whereas catechin derivatives such as gallocatechin (GC) and epigallocatechin (EGC) were present in GT extract. Moreover, theaflavin-3-O-gallate was observed in BT extract. Many of the catechin and its derivatives detected in the tea extracts were not identified in the tea yogurt samples. However, new phenolic compounds were present in GT-yogurt (i.e., kaempferol-3-rutinoside and quinic acid conjugate) but absent in GT extract. GT, WT, & BT extracts could be used to enriched-yogurt with phenolic compounds, which may have antioxidant properties.


Functional characterisation of romeharsha and clint1 reaffirms the link between plasma membrane homeostasis, cell size maintenance and tissue homeostasis in developing zebrafish epidermis.
In vertebrates, early developing epidermis is a bilayered epithelium consisting of an outer periderm and the underlying basal epidermis. It eventually develops into a multi-layered epithelium. The mechanisms that control the architecture and homeostasis of early developing bilayered epidermis have remained poorly understood. Recently, we have shown that the function of Myosin Vb, an actin based molecular motor, is essential in peridermal cells for maintenance of plasma membrane homeostasis. Furthermore, our analyses of the goosepimples/myosin Vb mutant unravelled a direct link between plasma membrane homeostasis, cell size maintenance and tissue homeostasis in the developing epidermis. However, it remained unclear whether this link is specific to myosin Vb mutant or this is a general principle. Here we have identified two more genetic conditions, romeharsha mutant and clint1 knockdown, in which membrane homeostasis is perturbed, as evident by increased endocytosis and accumulation of lysosomes. As a consequence, peridermal cells exhibit smaller size and increased proliferation. We further show that decreasing endocytosis in romeharsha mutant and clint1 morphants rescues or mitigates the effect on cell size, cell proliferation and morphological phenotype. Our data confirms generality of the principle by reaffirming the causal link between plasma membrane homeostasis, cell size maintenance and tissue homeostasis.


Hypoxia-inducible factors in OSCC.
Oral squamous cell carcinoma (OSCC) is a characteristic locally aggressive tumor in which hypoxia levels are very high, causing a low response to chemotherapy and providing basic resistance to anticancer drugs. Tumoral hypoxia directly depends on hypoxia-inducible factors (HIF). The goal of this paper is to describe HIF basic biology and tumor cells (HIF-1α, mainly), analyzing the effects of its expression in OSCC, study its relation with other molecules such as nitric oxide (NO), carbonic anhydrase (CA) or VEGF and assess the possibility of its manipulation as a therapeutic target.


Free glycine accelerates the autoproteolytic activation of human asparaginase.
Human asparaginase 3 (hASNase3), which belongs to the N-terminal nucleophile hydrolase superfamily, is synthesized as a single polypeptide that is devoid of asparaginase activity. Intramolecular autoproteolytic processing releases the amino group of Thr168, a moiety required for catalyzing asparagine hydrolysis. Recombinant hASNase3 purifies as the uncleaved, asparaginase-inactive form and undergoes self-cleavage to the active form at a very slow rate. Here, we show that the free amino acid glycine selectively acts to accelerate hASNase3 cleavage both in vitro and in human cells. Other small amino acids such as alanine, serine, or the substrate asparagine are not capable of promoting autoproteolysis. Crystal structures of hASNase3 in complex with glycine in the uncleaved and cleaved enzyme states reveal the mechanism of glycine-accelerated posttranslational processing and explain why no other amino acid can substitute for glycine.


Comparative physiology of burned and unburned Rhus laurina after chaparral wildfire.
Laurel Sumac (Rhus laurina) is a dominant member of the coastal chaparral community of southern California that survives periodic burning by wildfires by resprouting from a lignotuber (root crown). We investigated the physiological basis for resprouting by comparing shoot elongation, leaf nitrogen content, tissue water status, leaf conductance to water vapor diffusion, and photosynthetic rates of post-fire R. laurina to those of adjacent unburned shrubs. Resprouts had higher rates of shoot elongation, leaf conductance, and photosynthesis than mature, unburned shrubs. Leaf nitrogen contents were elevated in burned shrubs even though their leaves developed interveinal chlorosis. A comparison of soil water potential to predawn water potential indicated that roots of R. laurina remain active below 2 m during the first summer drought after wildfire. Our results support the hypothesis that lignotubers not only contain dormant buds that develop into aerial shoots after wildfire but they also supply nutrient resources that enhance shoot elongation. Because R. laurina is relatively sensitive to drought, yet very successful in its rapid recovery after fire, maintaining an active root system after shoot removal may be the primary function of the massive lignotuber formed by this species.


[Madelung's deformity - Why choose the Sauvé-Kapandji osteotomy?]
For bilateral Madelung's deformity in a 14-year-old girl we did the Sauvé and Kapandji operation without additional radius osteotomy. The operation was justified by the protrusion of the head of the ulna, the limitation of the rotation and diminution of the strength of the wrist, and inability to do sporting activity. The clinical findings were referred to the instability of the inferior radio ulnar joint There was no pain. The post operative review showed no pain, with recovery of grasp and of sporting activity (gymnastic and dance). The esthetic result satisfied the surgeon and the patient The operation permitted repositioning of the inferior radio ulnar joint. We have not seen synostosis of the osteotomy and the width of it increased progressively without radiological and physical instability of the inferior radio ulnar joint. The literature study allows us to discuss the different treatment possibilities and to explain our choice.The simplicity of this operation is interesting, because it was possible to stabilize the carpal joint, with abolition of the physical problems of this youg girl, without the needing osteotomy of the radius, which takes longer to recover, in those cases without pain before the operation.


Bis[2,4-di-bromo-6-(N-{4-[(E)-1-(benzyl-oxy-imino)-eth-yl]phen-yl}carboximido-yl)phenolato]copper(II).
In the title complex, [Cu(C22H17Br2N2O2)2], the Cu(II) ion is four-coordinated in a trans-CuN2O2 square-planar geometry by two phenolate O and two imino N atoms from two deprotonated N,O-bidentate ligands. In the crystal, the packing of the mol-ecules is controlled by C-H⋯π and π-π inter-actions [centroid-centroid distances = 3.568 (3), 3.678 (2), 3.717 (3) and 3.799 (2) Å] and weak Br⋯Br halogen bonds [3.508 (4) Å], linking the mol-ecules into an infinite three-dimensional supra-molecular network.


Molecular cloning of the restriction fragments derived from double EcoRI/PstI digestion of the calf satellite I DNA and their restriction analysis.
Calf satellite I DNA digested with EcoRI and PstI gives three fragments 643, 621 and 83 bp long. Two of them, the 643 and 621 bp were cloned using pBR 322 vector and analyzed by means of the MspI and HaeIII restriction enzymes.


Using real-time sound touch elastography to monitor changes in transplant kidney elasticity.
To use sound touch elastography (STE) to assess the changes of renal cortex among different complications following renal transplantation. A total of 31 patients with renal dysfunction after renal transplantation underwent an ultrasound-guided biopsy for pathological examination with conventional and STE ultrasound. The maximum elastic modulus (Emax) was determined, and the biopsy specimen was evaluated for evidence of significant differences among four different complications: drug-induced renal damage, acute rejection, chronic allograft nephropathy (CAN), and BK virus (BKV) nephropathy. Receiver operator characteristics were used to compare the diagnostic efficacy of STE ultrasound according to the pathological results. The quantitative index Emax of the STE technique was statistically significant among the four different complications (p<0.05). The distribution of the magnitude of Emax in the renal cortex was BKV nephropathy > CAN > acute rejection > drug induced renal damage. The renal cortex Emax was statistically different for the severity of renal fibrosis and tubular atrophy (p<0.05). Each of the four different complications of transplantation influenced the Emax of the renal cortex differently. Emax can be used to assess the severity of renal fibrosis and tubular atrophy.


Immunomodulatory potential of Nyctanthes abrortristis stem bark.
Phytotherapeutic modulation of the immune system to mitigate infectious ailments has been in vogue all over the world. The present work has been designed to scientifically explore the immunomodulatory potential of Nyctanthes arbortristis stem bark using mice models. Methanolic (MNA) and aqueous (ANA) extracts of N. arbortristis stem bark were evaluated for possible modulation in humoral immunity through serum immunoglobulin estimation. The variation in cellular immunity was assessed using neutrophil adhesion test, carbon clearance assay, and cyclophosphamide-induced neutropenia. Administration of MNA and ANA (both at 200 mg/kg, p.o.) significantly augmented the levels of serum immunoglobulins (humoral antibody), neutrophil adhesion, and phagocytic index (a measure of carbon clearance). Extracts also guarded the animals against cyclophosphamide-induced leukopenia, especially neutropenia. Results indicate that cellular and humoral immune responses were aroused by pretreatment of the animal with methanol and aqueous extract of N. arbortristis. Thus, the methanol and aqueous extract of N. arbortristis stem bark possesses a significant immunostimulant activity and can be used to uplift the immune system in the infectious condition.


Decompensated heart failure in the setting of kidney dysfunction: a community-wide perspective.
Patients with heart failure (HF) and kidney disease have a poor long-term outlook which has provided impetus for the identification of factors of prognostic importance and more fully understanding the impact of kidney dysfunction in patients with HF. Our objectives were to describe the characteristics, hospital treatment practices, as well as hospital and long-term outcomes in patients with varying degrees of kidney dysfunction who were hospitalized with acute HF at all medical centers in a large New England metropolitan area. Residents of the Worcester metropolitan area hospitalized with clinical findings of decompensated HF at 11 greater Worcester medical centers during 1995 and 2000 comprised the study sample. Kidney function was classified into 4 categories of estimated glomerular filtration rate (eGFR) for purposes of analysis: <30 (n = 569), 30-44 (n = 725), 45-59 (n = 763), and > or =60 (n = 2,293) ml/min per 1.73 m(2). The average age of the study sample was 76 years and 57% were women. Patients with severe kidney dysfunction were less likely to receive angiotensin-converting enzyme inhibitors, diuretics and digoxin during hospitalization for acute HF compared to patients with more normal kidney function. Patients with lower eGFR levels had higher in-hospital and post-discharge death rates in comparison to those with higher levels of eGFR. Our results demonstrate the impact of renal impairment on the prognosis of patients with decompensated HF. Our findings highlight the less than optimal management of these high-risk patients. Increased surveillance and enhanced treatment of patients with HF and kidney dysfunction remains warranted to improve the survival outlook of these patients.


Extensive scrotal lymphangioma circumscriptum: A case report with successful treatment with staged CO2 laser ablation.
Scrotal lymphangioma circumscriptum (LC) is an uncommon, benign skin disorder with multiple treatment modalities. We present staged treatment of extensive LC of the scrotum using staged carbon dioxide (CO2) laser ablation. We achieved near complete resolution of lesions with excellent cosmesis in the span of four months, from presentation to treatment. Therefore, the use of staged CO2 laser ablation should be strongly considered when LC presents in the genitourinary (GU) system, as it can lead to good cosmetic outcomes and prompt symptomatic relief in a short period of time.


The Evolution of Cerebral Language Localization: Historical Analysis and Current Trends.
Language localization has been an evolving concept over the past 150 years, with the emergence of several important yet conflicting ideologies. The classical theory, starting from the phrenologic work of Gall to the identification of specific regions of language function by Broca, Wernicke, and others, proposed that discrete subcomponents of language were organized into separate anatomic structural regions. The holism theory was postulated in an attempt to disclose that language function was instead attributed to a larger region of the cortex, in which cerebral regions may have the capability of assuming the function of damaged areas. However, this theory was largely abandoned in favor of discrete structural localizationist viewpoints. The subsequent cortical stimulatory work of Penfield led to the development of maps of localization, assigning an eloquent designation to specific regions. The expanding knowledge of cortical and subcortical anatomy allowed for the development of anatomically and functionally integrative language models. In particular, the dual stream model revisited the concept of regional interconnectivity and expanded the concept of eloquence. Advancements in cortical-subcortical stimulation, neurophysiologic monitoring, magnetic resonance diffusion tensor imaging/functional magnetic resonance imaging, awake neurosurgical technique, and knowledge gained by white matter tract anatomy and the Human Connectome Project, shed new light on the dynamic interconnectivity of the cerebrum. New studies are progressively opening doors to this paradigm, showing the dynamic and interdependent nature of language function. In this review, the evolution of language toward the evolving paradigm of dynamic language function and interconnectivity and its impact on shaping the neurosurgical paradigm are outlined.


Testicular histology in the adolescent with a varicocele.
A varicocele is a common cause of adult male infertility, and surgical ligation is a generally accepted mode of therapy. However, the guidelines for management of the adolescent with a varicocele are not clearly defined. Herein, we describe histologic abnormalities noted on testis biopsy in nine of 24 boys with moderate to large varicoceles. Surgical ligation of the varicocele is recommended when there is volume loss of the testis ipsilateral to the varicocele. All others should be reexamined periodically, and an attempt should be made to obtain a baseline semen analysis.


Free-breathing whole-heart coronary MRA with 3D radial SSFP and self-navigated image reconstruction.
Respiratory motion is a major source of artifacts in cardiac magnetic resonance imaging (MRI). Free-breathing techniques with pencil-beam navigators efficiently suppress respiratory motion and minimize the need for patient cooperation. However, the correlation between the measured navigator position and the actual position of the heart may be adversely affected by hysteretic effects, navigator position, and temporal delays between the navigators and the image acquisition. In addition, irregular breathing patterns during navigator-gated scanning may result in low scan efficiency and prolonged scan time. The purpose of this study was to develop and implement a self-navigated, free-breathing, whole-heart 3D coronary MRI technique that would overcome these shortcomings and improve the ease-of-use of coronary MRI. A signal synchronous with respiration was extracted directly from the echoes acquired for imaging, and the motion information was used for retrospective, rigid-body, through-plane motion correction. The images obtained from the self-navigated reconstruction were compared with the results from conventional, prospective, pencil-beam navigator tracking. Image quality was improved in phantom studies using self-navigation, while equivalent results were obtained with both techniques in preliminary in vivo studies.


A P25/(NH4)xWO3 hybrid photocatalyst with broad spectrum photocatalytic properties under UV, visible, and near-infrared irradiation.
In this study, a series of hybrid nanostructured photocatalysts P25/(NH4)xWO3 nanocomposites with the average crystallite size of P25 and (NH4)xWO3 of the sample was calculated to be about 30 nm and 130 nm, were successfully synthesized via a simple one-step hydrothermal method. The as-obtained samples was characterized by transmission electron microscopy (TEM), which implies that the P25/(NH4)xWO3 nanocomposites are fabricated with favourable nanosizd interfacial. The XPS results confirmed that the obtained sample consists of mixed chemical valences of W5+ and W6+, the low-valance W5+ sites could be the origin of NIR absorption. As revealed by optical absorption results, P25/(NH4)xWO3 nanocomposites possess high optical absorption in the whole solar spectrum of 200-2500 nm. Benefiting from this unique photo-absorption property and the synergistic effect of P25 and (NH4)xWO3, broad spectrum response photocatalytic activities covering UV, visible and near infrared regions on degradation of Rhodamine B have been realized by P25/(NH4)xWO3 nanocomposites. Meanwhile, the stability of photocatalysts was examined by the XRD and XPS of the photocatalysts after the reaction. The results show that P25/(NH4)xWO3 photocatalysts has a brilliant application prospect in the energy utilization to solve deteriorating environmental issues.


Adenosine kinase inhibitors attenuate opiate withdrawal via adenosine receptor activation.
Previous studies have demonstrated a role for adenosine in mediating opiate effects. This study examines the effects of indirect activation of adenosine receptors, via treatment with adenosine kinase inhibitors, on the expression of opiate withdrawal in mice. Mice receive chronic morphine treatment via implantation of subcutaneous morphine pellets (75 mg) for 72 h. Mice then receive parenteral treatment with adenosine kinase inhibitors, either 5'-amino-5'-deoxyadenosine (2, 5, 20, 40 mg/kg, intraperitoneal or i.p.) or iodotubericidin (1, 2, 5 mg/kg, i.p.), followed by naloxone injection and opiate withdrawal signs are measured over 20 min. Both adenosine kinase inhibitors significantly reduce the following opiate withdrawal signs in a dose-dependent manner compared to vehicle: withdrawal jumps, teeth chattering, forepaw tremors, and forepaw treads. Additionally, 5'-amino-5'-deoxyadenosine significantly reduces withdrawal-induced diarrhea and weight loss. Effects of 5'-amino-5'-deoxyadenosine (40 mg/kg) on opiate withdrawal signs appear to be mediated via adenosine receptor activation as they are reversed by pretreatment by adenosine receptor antagonist caffeine (20 mg, i.p.) but not by selective phosphodiesterase inhibitor Ro 20-1724 (10 mg/kg, i.p.). Adenosine receptor activation via adenosine kinase inhibitor treatment attenuates opiate withdrawal and these agents may be generally useful in the treatment of drug withdrawal syndromes.


Oncostatin M-induced upregulation of SDF-1 improves Bone marrow stromal cell migration in a rat middle cerebral artery occlusion stroke model.
Bone marrow-derived mesenchymal stem cells (BMSCs) exhibit potential regenerative effects on the injured brain. However, these effects are constrained by their limited ability to migrate to the injured site. Oncostatin M (OSM) has been shown to affect the proliferation and migration of mesenchymal stem cells. Therefore, in the present study, we explored whether OSM improves BMSC migration and secretion of growth factors and cytokines in a rat middle cerebral artery occlusion (MCAO) stroke model. The effect of OSM on the proliferation and apoptosis of rat BMSCs was first assessed in vitro, and the gene and secretion levels of factors related to cell nutrition and migration, such as SDF-1 and VEGF, were detected. To further explore underlying pathways triggered by OSM, BMSCs were treated with OSM in the presence or absence of inhibitors of the STAT3 and ERK pathways. Effects of OSM on SDF-1 expression in astrocytes and BMSC migration were also evaluated. In the rat MCAO model, OSM secretion levels were detected in the brain for up to 72 h after model establishment. Ventricle injection of OSM alone or OSM combined with caudal vein graft of BMSCs was then performed in MCAO stroke rats. After 72 h, production of SDF-1 and grafted BMSCs was detected in the lesion areas of the brain, and the nerve function score was evaluated. We found that the production of OSM continually increased in the brains of MCAO rats from 12 h to 72 h. OSM significantly upregulated SDF-1 in BMSCs via the STAT3 and ERK pathways and significantly promoted the expression of VEGF and MMP-2. OSM also promoted the secretion of SDF-1 in astrocytes through the STAT3 and ERK pathways to in turn enhance BMSC migration. Combination treatment with OSM and BMSCs in MCAO rats increased the migration efficiency of BMSCs in the brain, which significantly improved neurofunctional recovery while reducing the expression of inflammatory mediators and promoting the secretion of nutrition factors. Overall, these results show that OSM is highly expressed in the brains of MCAO stroke rats and can upregulate SDF-1 to promote BMSC migration. Thus, combination treatment with OSM and BMSCs improves the graft efficiency of BMSCs and neurofunctional recovery.


White paper on Ivermectin as a potential therapy for COVID-19.
A group of senior doctors with vast clinical experience met on 19th July'20 under the aegis of Academy of Advanced Medical Education. The panel looked at Ivermectin, one of the old molecule and evaluated it's use in COVID 19 (Novel Coronavirus Disease 2019) management. After critical panel discussion, all the attending doctors came to a conclusion that Ivermectin can be a potential molecule for prophylaxis and treatment of people infected with Coronavirus, owing to its anti-viral properties coupled with effective cost, availability and good tolerability and safety.


Surgical management of cecal diverticulitis.
Ninety surgical cases of cecal diverticulitis at the University of Hawaii hospitals were reviewed from 1980 to 1991. Seventy-eight percent of the study group were of Asian descent, with a mean age of 41.7 years. Right lower quadrant pain and tenderness were the only constant findings, occurring in 86 and 87 of the 90 patients, respectively. The most common preoperative diagnosis was acute appendicitis, occurring in 73% of patients. A right colectomy or cecectomy was performed in 49 patients, an appendectomy in 29, and a diverticulectomy in 10. Seventeen complications occurred, only 1 of which was in the appendectomy group. Follow-up of up to 10 years was successful in 27 of 29 appendectomy patients, only 4 of whom had recurrent pain. There were no instances of a missed cecal carcinoma. We concluded that in those patients in whom carcinoma can be ruled out and in whom there is no evidence of abscess formation, appendectomy combined with postoperative antibiotics is a safe and effective method for the treatment of cecal diverticulitis.


3D printed and spiral lithographically patterned erbium-doped polymer micro-waveguide amplifiers.
Infrared (IR)-emitting RE doped materials have been extensively used to fabricate active components of integrated optical devices in various fields, such as fiber amplifiers, telecommunications, optoelectronics, and waveguides. Among various RE elements, trivalent erbium ions (Er 3+) are of great interest since their emissive behavior span the low loss telecommunication window of 1300-1650 nm. In this paper, we report two types of polymeric waveguide amplifiers. 8 cm long, lithographically patterned spiral waveguides provide 8 dB of gain using a 980 nm pump power of 95 mW. Gain is observed from 1530 to 1590 nm. We further report the first demonstration of polymeric waveguide amplifiers fabricated using 3D printing methods based on two-photon lithography, paving the way for rapid prototyping of active 3D printed devices and active photonic devices which may transcend planar limitations.


Measurement of adrenolutin as an oxidation product of catecholamines in plasma.
Using the reverse phase high-performance liquid chromatography (HPLC) with mobile phases composed of simple acids, we have developed an assay technique for the measurement of adrenolutin, one of the oxidation products of catecholamines, in rat plasma. Ion-pairing chromatography permits the separation and quantitation of plasma adrenolutin (microM) in a linear manner. Sample preparation involved the precipitation of plasma proteins with perchloric acid and it is easier to handle a large number of samples at a time. However, we were unable to demonstrate the presence of adrenochrome, another oxidation product of catecholamines, in plasma since adrenochrome was rapidly destroyed in acid as well as in blood and was quickly changed into adrenolutin. Adrenolutin peak in HPLC was confirmed by 1) the retention time; 2) co-injection of adrenolutin and; 3) the appearance of 3H-adrenolutin after injection of 3H-norepinephrine. Administration of different catecholamines as well as adrenochrome and adrenolutin in rats also increased the level of adrenolutin in plasma. Adrenolutin was found to be present in plasma in other species including dog, rabbit and pig. High level of adrenolutin, which may represent total concentration of aminolutin in plasma, suggests the presence of an efficient mechanism for the oxidation of catecholamines under in vivo conditions.


The effects of attention on perceptual implicit memory.
Reports on the effects of dividing attention at study on subsequent perceptual priming suggest that perceptual priming is generally unaffected by attentional manipulations as long as word identity is processed. We tested this hypothesis in three experiments by using the implicit word fragment completion and word stem completion tasks. Division of attention was instantiated with the Stroop task in order to ensure the processing of word identity even when the participant's attention was directed to a stimulus attribute other than the word itself. Under these conditions, we found that even though perceptual priming was significant, it was significantly reduced in magnitude. A stem cued recall test in Experiment 2 confirmed a more deleterious effect of divided attention on explicit memory. Taken together, our findings delineate the relative contributions of perceptual analysis and attentional processes in mediating perceptual priming on two ubiquitously used tasks of word fragment completion and word stem completion.


Prevalence and Risk Factors Associated With Pelvic Rod/Screw Radiographic Lucency Following Scoliosis Surgery in Spastic Cerebral Palsy: A Longitudinal Study.
Radiographic lucency around a smooth pelvic rod (Galveston/unit rod technique) or sacroiliac/iliac screw following spinal fusion in children with nonambulatory spastic cerebral palsy (CP) has been described as a "windshield wiper" phenomenon. We evaluated demographics, radiographs, and complications in 101 cases from a single center to determine prevalence, risk factors, and complications associated with persistent radiographic lucency from 1 to 5 years following spinal fusion. Inclusion criteria were diagnosis of nonambulatory spastic quadriplegic CP [Gross Motor Function Classification System (GMFCS) IV-V], under 18 years of age, scoliosis treated by posterior fusion from upper thoracic to sacrum with pelvic fixation (Galveston rod, iliac screw, or sacroiliac screw), adequate radiographs (preoperative, immediate postoperative, first-year, and second-year), and minimum 5-year follow-up. We evaluated demographics, radiographic parameters, comorbidities, scoliosis curve type, type of pelvic screw/rod, use of off-set connector, screw width, associated with posterior column osteotomy and/or additional anterior spinal release concurrent with posterior spine fusion, and infection over the follow-up period. Specific attention was given to the area and shape of the radiographic lucency. The logistic regression analysis was performed for continuous and categorical variables to define risk factors ( P =0.05). In 101 patients, data were collected at mean intervals of 1-year, 2-year, and >5-year follow-up and were 12.9±1.5, 25.8±2.5, and 81.5±23.0 months, respectively. Prevalence of pelvic rod/screw radiographic lucency was unchanged at 33%, 35%, and 24% at 1-year, 2-year, and >5-year follow-up, respectively, and radiographic parameters did not change ( P >0.05). Furthermore, no risk factors or complications were associated with radiographic lucency around pelvic rods/screws ( P >0.05). In patients with spastic nonambulatory CP who had scoliosis treated with posterior spinal fusion from upper thorax to pelvis, the prevalence of pelvic rod/screw lucency is high. Persistent lucency >2 mm around pelvic implants is not clinically significant, does not warrant advanced imaging, or indicate a complication if stable over time and wider distally than proximally. Level III.


[Theory and critical aspects of the so-called steal effect of coronary dilators of the dipyridamole type].
The stenocardias appearing in the dipyridamol test are explained by the hypothesis of a steal effect. This notion is, concerning its contents, to be demarcated against other regulatory processes. With the help of a simple model can be made clear that the thesis of a steal effect stands in contradiction to haemodynamic basic conceptions and cannot be used for the explanation of the paradoxical effect of dipyridamol.


Estimation of pneumonitis risk in three-dimensional treatment planning using dose-volume histogram analysis.
Investigations to study correlations between the estimations of biophysical models in three dimensional (3D) treatment planning and clinical observations are scarce. The development of clinically symptomatic pneumonitis in the radiotherapy of thoracic malignomas was chosen to test the predictive power of Lyman's normal tissue complication probability (NTCP) model for the assessment of side effects for nonuniform irradiation. In a retrospective analysis individual computed-tomography-based 3D dose distributions of a random sample of 46/20 patients with lung/esophageal cancer were reconstructed. All patients received tumor doses between 50 and 60 Gy in a conventional treatment schedule. Biological isoeffective dose-volume histograms (DVHs) were used for the calculation of complication probabilities after applying Lyman's and Kutcher's DVH-reduction algorithm. Lung dose statistics were performed for single lung (involved ipsilateral and contralateral) and for the lung as a paired organ. In the lung cancer group, about 20% of the patients (9 out of 46) developed pneumonitis 3-12 (median 7.5) weeks after completion of radiotherapy. For the majority of these lung cancer patients, the involved ipsilateral lung received a much higher dose than the contralateral lung, and the pneumonitis patients had on average a higher lung exposure with a doubling of the predicted complication risk (38% vs. 20%). The lower lung exposure for the esophagus patients resulted in a mean lung dose of 13.2 Gy (lung cancer: 20.5 Gy) averaged over all patients in correlation with an almost zero complication risk and only one observed case of pneumonitis (1 out of 20). To compare the pneumonitis risk estimations with observed complication rates, the patients were ranked into bins of mean ipsilateral lung dose. Particularly, in the bins with the highest patient numbers, a good correlation was achieved. Agreement was not reached for the lung functioning as a paired organ. Realistic assessments for the prediction of radiation-induced pneumonitis seem to be possible. In this respect, the implementation of DVH-analysis in 3D planning could be a helpful tool for the evaluation of treatment plans.


Regional biomechanical and histological characterisation of the passive porcine urinary bladder: Implications for augmentation and tissue engineering strategies.
The aim of this study was to identify and quantify potential regional and directional variations in the quasistatic uniaxial mechanical properties of the passive urinary bladder wall. Overall, the lower body and trigone regions demonstrated the highest degree of directional anisotropy, whereas the ventral region demonstrated the least directional anisotropy. Significant regional anisotropy was found only along the apex-to-base direction. The dorsal and ventral regions demonstrated a significantly increased distensibility along the apex-to-base direction compared to the other bladder regions, whereas the trigone and lower body regions demonstrated the least distensibility. The trigone, lower body and lateral regions also demonstrated the highest tensile strength both at regional and directional levels. The study detected significant regional and directional anisotropy in the mechanical properties of the bladder and correlated this anisotropy to the distended and non-distended tissue histioarchitecture and whole organ mechanics. By elucidating the inhomogeneous nature of the bladder, the results from this study will aid the regional differentiation of bladder treatments in terms of partial bladder replacement with suitable natural or synthetic biomaterials, as well as the development of more realistic constitutive models of bladder wall biomechanics and improved computational simulations to predict deformations in the natural and augmented bladder.


Common patterns of corporate dermatology billing abuses in Medicare data sets.
Advances in technology have brought about significant changes in the way physicians' encounters are viewed. Patient charts once existed only in medical offices and hospital basements with medical billing done manually. Technologic changes have created new methods and expectations for oversight of the physician's practice. This contribution examines the effects of data transparency on dermatology. Medicare has also responded with new requirements for quality reporting. The ability to recreate clinical encounters from freely available and physician-identifiable claims data has led to lawsuits, investigations by news outlets, and distrust in the medical judgment of physicians. The release of data has also uncovered possible improper payments that have brought these practices out of the shadows. Medicare data over the past few years also show evidence of rapid corporatization and consolidation, which is changing the practice of dermatology.


[Oral presentation of non-Hodgkin's lymphoma. Literature review apropos of a clinical case].
The author makes a review of Non Hodgkin's Lymphoma and Oral Presentations, and includes a case report of the Oral and Maxilo-Facial Surgery of the Snt. António's Hospital--Porto.


Possible aspects of using cultured stem cell of laboratory primates in experimental medicine.
We describe the methods of isolation and culturing of mesenchymal stem cells from 3 monkey species Macaca mulatta, Papio hamadryas, and Macaca fascicularis. Flow cytofluorometry showed that the cells do not express CD34, CD45, and HLA-DR, but most of them (78-98%) express CD90 marker. The cardioprotective effects of cultured mesenchymal stem cells in cardiomyopathy induced by administration of antitumor anthracycline drugs (doxorubicin).


Dimethadione-induced fetotoxicity in rats.
The fetotoxic potential of dimethadione was studied in rats given single daily oral dosages of 0, 54, 433 or 541 mg/kg on days 1--21 or 6--15 of gestation. No maternal toxicity was observed following treatment on days 6--15. When administered from days 1 to 21 only the highest dose (541 mg/kg) produced a significant reduction in maternal body weight gain. Dimethadione caused a dose-related decrease in fetal weight and an increased incidence of umbilical hernia, ecchymoses and subcutaneous edema. There were also increased incidences of non-specific skeleton defects which consisted of unilateral or bilateral wavy ribs, additional ribs (14th rib, uni- and bilateral), retarded ossification of calvaria and a wide variety of sternal defects. Specific defects were bent radius and ulna, and bent tibia and fibula which increased with increasing dosages of dimethadione. Fetal mortality and incidence of skeletal anomalies were higher when the treatment was given on days 1--21 of gestation than on days 6--15 of pregnancy.


Cleavage from the N-terminal region of beta Bp crystallin during aging of the human lens.
Polyclonal antisera have been made to synthetic peptides that correspond to the N-terminal (residues 1-12) and C-terminal (residues 195-204) sequences of bovine beta Bp crystallin. Both anti-beta Bp1-12 and anti-beta Bp195-204 recognize specifically the beta Bp component of bovine lens. In the young human lens, anti-beta Bp195-204 recognizes predominantly the 26,000 MW form of beta Bp, while in older lenses this same antiserum recognizes mainly the 22,000 MW in vivo proteolysis product. In contrast, during aging of the normal human lens anti-beta Bp1-12 recognizes only decreasing amounts of the 26,000 MW form of beta Bp, with no binding to the 22,000 MW form of this polypeptide. These results suggest that during aging of the normal human lens, the N-terminus of beta Bp is the preferred site of in vivo proteolysis.


EBV promotes vascular mimicry of dormant cancer cells by potentiating stemness and EMT.
Vascular mimicry (VM) is defined as a vascular channel-like structure composed of tumor cells that correlates with the growth of cancer cells by providing blood circulation. However, whether VM can be formed in dormant cancer cells remains unclear. Our previous research revealed that polyploid giant cancer cells (PGCCs) are specific dormant cells related to the poor prognosis of head and neck cancer. Here, we demonstrated that EBV could promote VM formation by PGCCs in vivo and in vitro. Furthermore, we revealed that the activation of the ERK pathway partly mediated by LMP2A is responsible for stemness, and the acquisition of the stemness phenotype is crucial to the malignant biological behavior of PGCCs. The epithelial-to-mesenchymal transition (EMT) process plays a considerable role in PGCCs, and EMT progression is vital for EBV-positive PGCCs to form VM. This is the first study to reveal that EBV creates plasticity in PGCC-VM and provide a new strategy for targeted anti-tumor therapy.


Effects of climatic factors on the prevalence of influenza virus infection in Cheonan, Korea.
Big data can be used to correlate diseases and climatic factors. The prevalence of influenza (flu) virus, accounting for a large proportion of respiratory infections, suggests that the effect of climate variables according to seasonal dynamics of influenza virus infections should be investigated. Here, trends in flu virus detection were analyzed using data from 9,010 tests performed between January 2012 and December 2018 at Dankook University Hospital, Cheonan, Korea. We compared the detection of the flu virus in Cheonan area and its association with climate change. The flu virus detection rate was 9.9% (894/9,010), and the detection rate was higher for flu virus A (FLUAV; 6.9%) than for flu virus B (FLUBV; 3.0%). Both FLUAV and FLUBV infections are considered an epidemic each year. We identified 43.1% (n = 385) and 35.0% (n = 313) infections in children aged < 10 years and adults aged > 60 years, respectively. The combination of these age groups encompassed 78.1% (n = 698/894) of the total data. Flu virus infections correlated with air temperature, relative humidity, vapor pressure, atmospheric pressure, particulate matter, and wind chill temperature (P < 0.001). However, the daily temperature range did not significantly correlate with the flu detection results. This is the first study to identify the relationship between long-term flu virus infection with temperature in the temperate region of Cheonan.


Visual optical biosensors based on DNA-functionalized polyacrylamide hydrogels.
Biosensors are devices that can provide quantitative or semi-quantitative analytical information about target molecules, where molecular recognition is based on biomolecular interactions. In recent years, DNA has emerged as a useful molecule for biosensor development since DNA can not only recognize its complementary strand, but also metal ions, small molecules, proteins and cells utilizing DNA aptamer technology. Converting DNA binding events into useful biosensors often require sensor immobilization. Among the various materials for sensor immobilization, hydrogels are particularly attractive. Hydrogels are crosslinked hydrophilic polymer networks that undergo swelling in water. In a gel, DNA immobilization can take place in 3D, allowing for high DNA loading capacity. Hydrogels are transparent, offering low optical background. The gel volume is affected by many environmental parameters such as temperature, pH, ionic strength, and solvent composition. In this paper, we present a concise summary of recent developments in DNA-functionalized hydrogel biosensors for visual detection. Detailed methods for immobilizing DNA biosensors in monolithic polyacrylamide gels and gel microparticles are supplied.


Measuring hand hygiene compliance rates at hospital entrances.
Despite the importance of hand hygiene in the health care setting, there are no studies evaluating hand hygiene compliance at hospital entrances. The study was prospectively performed over a 33-week period from March 30, 2014-November 15, 2014, to evaluate hand hygiene compliance in 2 hospital reception areas. We compared electronic handwash counters with the application of radiofrequency identification (GOJO SMARTLINK) (electronic observer) that counts each activation of alcohol gel dispensers to direct observation (human observer) via remote review of video surveillance. We found low hand hygiene compliance rates of 2.2% (99/4,412) and 1.7% (140/8,277), respectively, at reception areas A and D, detected by direct observation. Using the electronic observer, we measured rates of 17% (15,624/91,724) and 7.1% (51,605/730,357) at reception areas A and D, respectively. For the overall time period of simultaneous electronic and human observation, the human observer captured 1% of the hand hygiene episodes detected by the electronic observer. Our study showed very low hand hygiene compliance in hospital reception areas, and we found an electronic hand hygiene system to be a useful method to monitor hand hygiene compliance.


Interstimulus interval dependence of the auditory vertex response and its magnetic counterpart: implications for their neural generation.
Auditory vertex responses elicited by short tone bursts were compared with their magnetic counter parts. Special attention was paid to the behaviour of the N100 deflection of the response. Electrical responses were recorded from scalp locations Fp2, Fz, Cz, Pz, C4 and T4 and the magnetic responses half way between P4 and T6, at a point where the response has one of its amplitude extrema. Different ISIs (from 1 to 16 sec) were applied in order to differentiate specific and nonspecific evoked potential components from each other. The main results were as follows: (1) The scalp distsribution of the electical vertex response depends on the ISI used: with frequent stimulation there are no marked differences in the amplitudes of N100 between frontal and central areas but with long ISIs the amplitude maxima move to the vertex. (2) The magnetic responses also show a clear ISI dependence. The magnetic counterpart of N100 saturates at shorter ISIs than N100 recorded from the vertex. Independent of the ISI the magnetic counterpart of P200 is constantly very small. On the basis of the different sensitivities of the EEG and MEG to current sources of different orientations it is concluded that the auditory vertex response contains both modality specific and non-specific components. Experimental conditions, especially the ISI used, determine the relative contributions of these components to the potential recorded on the scalp.


Occipital nerve block is effective in craniofacial neuralgias but not in idiopathic persistent facial pain.
Occipital nerve block (ONB) has been used in several primary headache syndromes with good results. Information on its effects in facial pain is sparse. In this chart review, the efficacy of ONB using lidocaine and dexamethasone was evaluated in 20 patients with craniofacial pain syndromes comprising 8 patients with trigeminal neuralgia, 6 with trigeminal neuropathic pain, 5 with persistent idiopathic facial pain and 1 with occipital neuralgia. Response was defined as an at least 50% reduction of original pain. Mean response rate was 55% with greatest efficacy in trigeminal (75%) and occipital neuralgia (100%) and less efficacy in trigeminal neuropathic pain (50%) and persistent idiopathic facial pain (20%). The effects lasted for an average of 27 days with sustained benefits for 69, 77 and 107 days in three patients. Side effects were reported in 50%, albeit transient and mild in nature. ONBs are effective in trigeminal pain involving the second and third branch and seem to be most effective in craniofacial neuralgias. They should be considered in facial pain before more invasive approaches, such as thermocoagulation or vascular decompression, are performed, given that side effects are mild and the procedure is minimally invasive.


Considering Organismal Physiology in Laboratory Studies of Rodent Behavior.
Any experiment conducted in a rodent laboratory is done so against the backdrop of each animal's physiological state at the time of the experiment. This physiological state can be the product of multiple factors, both internal (e.g., animal sex, strain, hormone cycles, or circadian rhythms) and external (e.g., housing conditions, social status, and light/dark phases). Each of these factors has the potential to influence experimental outcomes, either independently or via interactions with others, and yet there is little consistency across laboratories in terms of the weight with which they are considered in experimental design. Such discrepancies-both in practice and in reporting-likely contribute to the perception of a reproducibility crisis in the field of behavioral neuroscience. In this review, we discuss how several of these sources of variability can impact outcomes within the realm of common learning and memory paradigms.


Genetic polymorphisms and mutation rates of 27 Y-chromosomal STRs in a Han population from Guangdong Province, Southern China.
In this study, we collected blood samples from 1033 father-son pairs of a Han population from Guangdong Province, Southern China, of which 1007 fathers were unrelated male individuals. All together, 2040 male individuals were analyzed at 27 Y-chromosomal short tandem repeats (Y-STRs) with Yfiler(®) Plus system. A total of 1003 different haplotypes were observed among 1007 unrelated fathers, with the overall haplotype diversity (HD) 0.999992 and discrimination capacity (DC) 0.996. The gene diversity (GD) values for the 27 Y-STR loci ranged from 0.4400 at DYS438 to 0.9597 at DYS385a/b. 11 off-ladder alleles and 25 copy number variants were detected in 1007 males. Population relationships were analyzed by comparison with 19 other worldwide populations. With 27,920 allele transfers in 1033 father-son pairs, 124 mutation events occurred, of which 118 were one-step mutations and 6 were two-step mutations. Eleven father-son pairs were found to have mutations at two loci, while one pair at three loci. The estimated locus-specific mutation rates varied from 0 to 1.74×10(-2), with an average estimated mutation rate 4.4×10(-3) (95%CI: 3.7×10(-3) to 5.3×10(-3)). Mutations were most frequently observed at three rapidly mutating Y-STRs (RM Y-STRs), DYS576, DYS518 and DYS627. However, at DYS570, DYS449 and DYF387S1 loci, which were also described as RM Y-STRs, the mutation rates in Guangdong Han population were not as high as estimated in other populations.


The social construction of the patient: patients and illnesses in other ages.
The way individual conceptions and experiences of illnesses are socially constructed is shown through the historical analysis of diseases, particularly epidemics. For centuries, sickness did not clearly correspond to what we now call the patient status. Several factors were necessary for this social status to develop. Sickness had to cease being a mass phenomenon. Medicine had to become capable of providing efficacious treatment and of taking the sick out of the custody of religious institutions. Through social legislation, the notions of illness and of health had to be related to the world of work. Owing to the first two factors, sickness was transformed from a collective into an individual affair, from a way of dying into a way of living. Once related to production processes and medical custody, it became the individual's social condition.


Ochratoxin A impairs "postproximal" nephron function in vivo and blocks plasma membrane anion conductance in Madin-Darby canine kidney cells in vitro.
Ochratoxin A (OTA) is a widespread nephrotoxin which causes porcine nephropathy and is supposed to have caused the human Balkan endemic nephropathy. We performed experiments in vivo and in vitro to elucidate the mechanism of OTA action in renal epithelium. Application of OTA to male Wistar rats [1.25 mumol/(kg.day)] for 6 days led to a reduction of glomerular filtration rate (to 63% of control), an increased fractional water (194% of control), Na+ (199% of control), K+ (147% of control) and Cl- (270% of control) excretion and an increased dependence of the osmole clearance on urine flow. Acute application of OTA to rats (3 mumol/kg) increased urinary pH from 6.0 +/- 0.2 to 6.6 +/- 0.1 and urinary NaCl excretion, but decreased titratable acid excretion to 47% of control. As these in vivo findings may be the result of an action of OTA beyond the proximal tubule ("postproximal") we investigated the effect of OTA on cultured Madin-Darby canine kidney (MDCK) cells, regarded as a model of collecting duct epithelium. In confluent monolayers formed by MDCK cells OTA reduced the number of domes in a dose-dependent manner and impaired the formation of a transepithelial Cl- gradient. Electrophysiological measurements in giant MDCK cells revealed that OTA blocks fractional anion conductance of the plasma membrane with an IC50 value of 30 +/- 5 nmol/l, unmasking OTA as a naturally occurring anion conductance blocker about 20-times more effective than the most potent synthetic blocker 5-nitro-2-(3-phenylpropyl-amino) benzoic acid (NPPB) (IC50 = 600 +/- 50 nmol/l).(ABSTRACT TRUNCATED AT 250 WORDS)


Ovarian function in Nelore (Bos taurus indicus) cows after post-ovulation hormonal treatments.
Maternal recognition of pregnancy in the cow requires successful signaling by the conceptus to block luteolysis. Conceptus growth and function depend on an optimal uterine environment, regulated by luteal progesterone. The objective of this study was to test strategies to optimize luteal function, as well as prevent a dominant follicle from initiating luteolysis. Nelore (Bos taurus indicus) beef cows (n=40) were submitted to a GnRH/PGF(2alpha)/GnRH protocol. Cows that ovulated from a dominant ovarian follicle (ovulation=Day 0) were allocated to receive: no additional treatment (G(C); n=7); 3000IU of hCG on Day 5 (G(hCG); n=5); 5mg of estradiol-17beta on Day 12 (G(E2); n=6); or 3000IU of hCG on Day 5 and 5mg of estradiol-17beta on Day 12 (G(hCG/E2); n=5). Ultrasonographic imaging of the ovaries, assessment of plasma progesterone concentration, and detection of estrus were done daily from Day 5 to the day of subsequent ovulation. Treatment with hCG induced an accessory CL, increased CL volume, and plasma progesterone concentration throughout the luteal phase (P<0.01). Estradiol-17beta induced atresia and recruitment of a new wave of follicular growth; it eliminated a potentially estrogen-active, growing ovarian follicle within the critical period for maternal recognition of pregnancy, but it also hastened luteolysis (Days 16 or 17 vs. Days 18 or 19 in non-treated cows). In conclusion, the approaches tested enhanced luteal function (hCG) and altered ovarian follicular dynamics (estradiol-17beta), but were unable to extend the life-span of the CL in Nelore cows.


Anti-proliferative properties of Clozic, a disease-modifying anti-arthritic agent.
Cytostatic drugs have been used in the treatment of rheumatoid arthritis but are of limited clinical application due to their severe toxic side-effects. We have discovered that 'Clozic' (ICI 55897), an agent with disease-modifying properties in rheumatoid arthritis patients, inhibits the growth of a variety of mammalian cell types including a matrix-secreting cell culture derived from neonatal rat hearts. The inhibition of growth was reversible and no loss of cell viability occurred when measured by lactate dehydrogenase released into the medium or by vital staining, suggesting a cytostatic rather than a cytotoxic mechanism. Cytostatic activity was observed at ICI 55897 concentrations within the reported therapeutic plasma concentration range and was related to the concentration of unbound compound, since the effect could be reduced by increasing the albumin concentration in the medium. Other oxyalkanoic acids inhibited cell growth. Their inhibitory potency correlated with lipophilicity. The anti-proliferative potencies of R and S enantiomers of two oxyalkanoic acids containing asymmetric centres were similar. These observations suggest that the anti-proliferative effect of the oxyalkanoic acids is due to their interaction with lipophilic cell target sites.


The effects of the financial crisis on primary prevention of cancer.
The present financial crisis will affect primary cancer prevention through several avenues: personal lifestyle choices, exposure to environmental risk factors, decisions made in the private sector and public policy on cancer prevention. Whilst it is clearly problematic to reach solid conclusions on a direct connection between economic crises and cancer mortality, we can identify trends that provide guidance for further action. For some lifestyle choices such as smoking or diet, we argue that public policy may channel existing tendencies during times of crisis for clear added value. In other areas, including research and health system investments, we will make the case that the resources not used now for cancer prevention efforts will lead to increased costs (both financial and human) down the road. Policy makers face a clear choice: they can follow a cost contention strategy, which may reduce expenditure in the short-term only to increase it in the long-term, or they can use the financial crisis as an opportunity to make difficult choices in terms of health service rationalisation, whilst at the same time strengthening evidence-based prevention policies. In short, we argue that despite the scarcity of funds and the governmental priorities on economic recovery, cancer prevention is more relevant now than ever.


A possible neurophysiological basis for psychological pain.
There is some evidence that the anterior cingulate cortex (ACC) contains nociceptive neurons, and other neurons which respond to conditioned aversive stimuli. It is not implausible that nociceptive input and conditioned aversive input converge onto common neurons in the ACC. Following from this hypothesis, the psychological pain which occurs in circumstances that arouse disgust or aversion may involve the same mechanisms that give rise to referred pain.


Tamoxifen-induced cytotoxicity in breast cancer cells is mediated by glucose-regulated protein 78 (GRP78) via AKT (Thr308) regulation.
Glucose regulated protein 78 (GRP78) has recently been suggested to be associated with drug resistance in breast cancer patients. However, the precise role of GRP78 in drug resistance and the involved signaling pathways are not clearly understood. In the present study, we show that among a panel of drugs, namely Paclitaxel (TAX), Doxorubicin (DOX), 5-fluorouracil (5-FU), UCN-01 and Tamoxifen (TAM) used, TAM alone up-regulated the expression of GRP78 significantly and induced apoptosis in MCF-7 and MDA-MB-231 cells. Interestingly, inhibition of GRP78 by a specific pharmacological inhibitor, VER-155008 augmented TAM-induced apoptosis, and overexpression of GRP78 rendered the cells resistant to TAM-induced cell death suggesting a role for GRP78 in TAM-induced cytotoxicity. Mechanistically, the expression of phosphorylated AKT as determined by Western blot analyses revealed that TAM selectively upregulated phosphorylation of AKT at Thr308 but not at Ser473, and siRNA silencing of GRP78 resulted in inhibition of AKT phosphorylation at Thr308 but not at Ser473. Further, a GRP78 inhibitor, VER155008 inhibited TAM-induced phosphorylation of GSK3β, a downstream substrate of AKT. These results, thus suggests a role for GRP78 in TAM-induced AKT activation. Additionally, co-localization studies by immunofluorescence, and immunoprecipitation experiments demonstrated a complex formation of AKT and GRP78. Furthermore, in glucose-free medium, the cells were sensitized to TAM-induced cell death that was associated with reduced AKT phosphorylation at Thr308, thus strengthening the association of AKT regulation with drug response. Collectively, our findings identify a role of GRP78 in AKT regulation in response to TAM in breast cancer cells.


Squamous cell carcinoma antigen: a role in the early identification of nodal metastases in men with squamous cell carcinoma of the penis.
To evaluate whether serum squamous cell carcinoma antigen (SCCAg) measurements may be of use in identifying nodal metastases in patients with SCC of the penis after treating the primary tumour. The levels of SCCAg were analysed in 11 men with penile SCC between 1994 and 2001. An elevated SCCAg level had a sensitivity of 57% (95% confidence interval, CI, 18-90%) and a specificity of 100% (CI 40-100%) for nodal metastases. Levels of SCCAg increased exponentially in patients who developed nodal metastases after treatment of the primary tumour, and were elevated before clinical or radiological evidence of nodal disease. Either the absolute level or the rate of rise of SCCAg may be a useful tool with which to follow patients after excision of the primary tumour. It may be more sensitive than computed tomography and magnetic resonance imaging in detecting recurrence, but further evaluation is needed.


The impact of frailty on ventral hernia repair outcomes in a statewide database.
Preoperative frailty is a strong predictor of postoperative morbidity in the general surgery population. Despite this, there are a paucity of research examining the effect of frailty on outcomes after ventral hernia repair (VHR), one of the most common abdominal operations in the USA. We examined the association of frailty with short-term postoperative outcomes while accounting for differences in preoperative, operative, and hernia characteristics. We retrospectively reviewed the Michigan Surgery Quality Collaborative Hernia Registry (MSQC-HR) for adult patients who underwent VHR between January 2020 and January 2022. Patient frailty was assessed using the validated 5-factor modified frailty index (mFI5) and categorized as follows: no (mFI5 = 0), moderate (mFI5 = 1), and severe frailty (mFI5 ≥ 2). Our primary outcome was any 30-day complication. Multivariable logistic regression was used to evaluate the association of frailty with outcomes while controlling for patient, operative, and hernia variables. A total of 4406 patients underwent VHR with a mean age (SD) of 55 (15) years, 2015 (46%) females, and 3591 (82%) white patients. The mean (SD) BMI of the cohort was 33 (8) kg/m2. A total of 2077 (47%) patients had no frailty, 1604 (36%) were moderately frail, and 725 (17%) were severely frail. The median hernia size (interquartile range) was 2.5 cm (1.5-4.0 cm). Severe frailty was associated with increased odds of any complication (adjusted Odds Ratio (aOR) 3.12, 95% CI 1.78-5.47), serious complication (aOR 5.25, 95% CI 2.17-13.19), SSI (aOR 3.41, 95% CI 1.58-7.34), and post-discharge adverse events (aOR 1.70, 95% CI 1.24-2.33). After controlling for patient, operative, and hernia characteristics, frailty was independently associated with increased odds of postoperative complications. These findings highlight the importance of preoperative frailty assessment for risk stratification and to inform patient counseling.


Rosetta Stone of NLR Innate Immunity.
The formation of NLR inflammasomes is a central step in the initiation of the innate immune response. Two recent publications describe the structure of the NAIP2-NLRC4 inflammasome and derive an elegant model of NLR inflammasome formation, whereby binding of the pathogen-molecule-bound NLR NAIP2 to NLRC4 leads to the activation of NLRC4 and initiation of self-propagating NLRC4 inflammasome formation.


Action potential and calcium dependence of tonic somatodendritic dopamine release in the Substantia Nigra pars compacta.
Despite the importance of somatodendritic dopamine (DA) release in the Substantia Nigra pars compacta (SNc), its mechanism remains poorly understood. Using a novel approach combining fast-scan controlled-adsorption voltammetry (FSCAV) and single-unit electrophysiology, we have investigated the mechanism of somatodendritic release by directly correlating basal (non-stimulated) extracellular DA concentration ([DA]out ), with pharmacologically-induced changes of firing of nigral dopaminergic neurons in rat brain slices. FSCAV measurements indicated that basal [DA]out in the SNc was 40.7 ± 2.0 nM (at 34 ± 0.5°C), which was enhanced by amphetamine, cocaine, and L-DOPA, and reduced by VMAT2 inhibitor, Ro4-1284. Complete inhibition of firing by TTX decreased basal [DA]out , but this reduction was smaller than the effect of D2 receptor agonist, quinpirole. Despite similar effects on neuronal firing, the larger decrease in [DA]out evoked by quinpirole was attributed to cell membrane hyperpolarization and greater reduction in cytosolic free Ca2+ ([Ca2+ ]in ). Decreasing extracellular Ca2+ also reduced basal [DA]out , despite increasing firing frequency. Furthermore, inhibiting L-type Ca2+ channels decreased basal [DA]out , although specific Cav 1.3 channel inhibition did not affect firing rate. Inhibition of sarcoplasmic/endoplasmic reticulum Ca2+ -ATPase (SERCA) also decreased [DA]out , demonstrating the importance of intracellular Ca2+ stores for somatodendritic release. Finally, in vivo FSCAV measurements showed that basal [DA]out in the SNc was 79.8 ± 10.9 nM in urethane-anesthetized rats, which was enhanced by amphetamine. Overall, our findings indicate that although tonic somatodendritic DA release is largely independent of action potentials, basal [DA]out is strongly regulated by voltage-dependent Ca2+ influx and release of intracellular Ca2+ . OPEN SCIENCE BADGES: This article has received a badge for *Open Materials* because it provided all relevant information to reproduce the study in the manuscript. The complete Open Science Disclosure form for this article can be found at the end of the article. More information about the Open Practices badges can be found at https://cos.io/our-services/open-science-badges/.


Psychometric Results of a New Patient-Reported Outcome Measure for Uveal Melanoma Post-Brachytherapy Treatment: The PROM-UM.
The objective of this study was to evaluate the psychometric properties of a new patient-reported outcome instrument intended for use with patients who have undergone brachytherapy for uveal melanoma (PROM-UM). Classical test theory and item response theory were used to evaluate the performance of individual items and domains. A convenience sample of 439 participants who had undergone brachytherapy for uveal melanoma from one of three North American ocular oncology treatment centers were included in this cross-sectional study. Exploratory factor analysis identified three domains which were labelled "Symptom Impairment", "Worry", and "Discomfort". The acceptability of the instrument was supported by little missing data (range = 0.00-1.14%) and low maximum endorsement (range = 0.00-1.82%). Item-total (range = 0.68-0.85) and inter-item (range = 0.74-0.80) correlations indicated acceptable reliability. Discrimination and difficulty were assessed using item response theory. Items in all three domains indicated moderate to very high discrimination (range = 1.00-4.10). Two items in the Symptom Impairment domain were too difficult to measure. Response ranges in the other two domains demonstrated acceptable difficulty. These results from the study indicate that this new patient-reported outcome instrument can be used with patients treated with brachytherapy for uveal melanoma. Providers could use this instrument to help inform post-treatment management.


Determination of parabens in wastewater samples via robot-assisted dynamic single-drop microextraction and liquid chromatography-tandem mass spectrometry.
Dynamic single-drop microextraction (SDME) was automatized employing an Arduino-based lab-made Cartesian robot and implemented to determine parabens in wastewater samples in combination with liquid chromatography-tandem mass spectrometry. A dedicated Arduino sketch controls the auto-performance of all the stages of the SDME process, including syringe filling, drop exposition, solvent recycling, and extract collection. Univariate and multivariate experiments investigated the main variables affecting the SDME performance, including robot-dependent and additional operational parameters. Under selected conditions, limit of detections were established at 0.3 µg/L for all the analytes, and the method provided linear responses in the range between 0.6 and 10 µg/L, with adequate reproducibility, measured as intraday relative standard deviations (RSDs) between 5.54% and 17.94%, (n = 6), and inter-days RSDs between 8.97% and 16.49% (n = 9). The robot-assisted technique eased the control of dynamic SDME, making the process more feasible, robust, and reliable so that the developed setup demonstrated to be a competitive strategy for the automated extraction of organic pollutants from water samples.