Inherited neuropathies with predominant upper limb involvement: genetic heterogeneity and overlapping pathologies.
In a subset of patients with inherited peripheral neuropathies the first symptom is atrophy and weakness of the intrinsic muscles of the hands, without involvement of lower limbs until later in the disease course. The exact pathomechanisms of this phenotype are currently unknown. The aim of this study was to characterize the clinical, neurophysiological and genetic features of a group of patients with a clinical diagnosis of upper limb predominant Charcot-Marie-Tooth disease (CMT). The clinical, electrophysiology and genetic data of 11 patients with upper limb predominant peripheral neuropathy selected from a single-centre cohort of 461 patients diagnosed with inherited neuropathy were analysed and the clinical, electrophysiological and genetic characteristics of these patients reported. An overlapping phenotype of neuropathy and myopathy was detected in two patients. Four patients carry autosomal dominant mutations in GARS and a single patient had a homozygous mutation in SH3TC2. However, the underlying genetic diagnosis could not be confirmed in six patients by gene panel sequencing. Upper limb-onset inherited neuropathies are genetically heterogeneous and, in some cases, there is an overlapping myopathy. Autosomal dominant GARS mutations are the most common genetic cause; however, mutations in other CMT genes may also result in this phenotype in individual patients. The majority of these patients cannot be genetically diagnosed by gene panel testing of known CMT and myopathy genes, suggesting further genetic heterogeneity and highlighting the importance of further genetic investigations in these patients and families.


Ovalbumin antibody-based fluorometric immunochromatographic lateral flow assay using CdSe/ZnS quantum dot beads as label for determination of T-2 toxin.
This work describes an anti-ovalbumin antibody-based lateral flow immunoassay (LFI) for T-2 toxin. The antibody uses a coating antigen as a bifunctional element for universality and introduces preincubation to improve the detection limits of the method. T-2 toxin and ovalbumin-modified T-2 toxin competitively binds on the anti-T-2 toxin monoclonal antibody modified on CdSe/ZnS quantum dot beads during preincubation. The modified T-2 toxin acts as a bifunctional element that forms immuno complexes during preincubation and combines with anti-ovalbumin antibody coated in the test line through the ovalbumin terminal. Fluorescence is detected at 610 nm on the test zone following photoexcitation at 365 nm. It has a reverse dose-effect relationship with the amount of T-2 toxin. The calibration plot is linear in the 20-110 fg mL-1 T-2 toxin concentration range, and the limit of detection (LOD) is 10 fg mL-1, which is lower by 8-fold than that of the traditional LFI system (LOD 80 fg mL-1) and one order of magnitude than those of LFIs with labels of colloidal gold nanoparticles (LOD 150 fg mL-1) or fluorophores (LOD 190 ng mL-1). Universality was verified through aflatoxin B1 detection using the established ovalbumin antibody-based LFI system (LOD 10 fg mL-1). The performance of the method was compared with that of established systems and a commercial ELISA kit (LOD 360 fg mL-1). Graphical abstractSchematic representation of ovalbumin antibody-based immunochromatographic lateral flow assay for T-2 toxin. Preincubation is introduced for high sensitivity. T-2- anti-ovalbumin acts as a bi-functional element for universality. CdSe/ZnS quantum dot beads act as label. Fluorometric signal is detected at 610 nm.


Neuroprotective effects of salidroside in the PC12 cell model exposed to hypoglycemia and serum limitation.
The hypoglycemia and serum limitation-induced cell death in cultured PC12 cells represents a useful in vitro model for the study of brain ischemia and neurodegenerative disorders. Salidroside is a phenylpropanoid glycoside isolated from Rhodiola rosea L., a traditional Chinese medicinal plant, and has displayed a broad spectrum of pharmacological properties. In this study, MTT assay, Hoechst 33342 staining, and flow cytometry with annexin V/PI staining collectively showed that pretreatment with salidroside attenuated, in a dose-dependent manner, cell viability loss, and apoptotic cell death in cultured PC12 cells induced by hypoglycemia and serum limitation. RT-PCR, Western blot analysis, and enzymatic colorimetric assay indicated the changes in expression levels of Bcl-2, Bax, and caspase3 in PC12 cells on exposure to hypoglycemia and serum limitation with and without salidroside pretreatment, respectively. Rhodamine 123 staining and flow cytometry with 2',7'-Dichlorofluorescin diacetate staining revealed the changes in the mitochondrial membrane potential and radical oxygen species (ROS) production in PC12 cells on exposure to hypoglycemia and serum limitation with and without salidroside pretreatment, respectively. The experimental results suggest that salidroside protects the PC12 cells against hypoglycemia and serum limitation-induced cytotoxicity possibly by the way of the modulation of apoptosis-related gene expression, the restoration of the mitochondrial membrane potential, and the inhibition of the intracellular ROS production. Our findings might raise a possibility of potential therapeutic applications of salidroside for preventing and treating cerebral ischemic and neurodegenerative diseases.


Intraperitoneal rupture of an infected urachal cyst: an unusual cause of acute abdomen in children. A case report and review of the literature.
Urachal cysts are rare and usually asymptomatic until they become infected. This is a report of a case of peritonitis due to intraperitoneal rupture of an infected urachal cyst in a 3-month-old girl. A review of the literature found 31 similar cases previously reported, only seven of which were described in children. The diagnosis and treatment of intraperitoneal perforation of an infected urachal cyst in a child are discussed.


Effect of an aqueous extract of Averrhoa carambola L. on endothelial function in rats with ventricular remodelling.
Ventricular remodelling leads to cardiomyocyte hypertrophy, myocardial fibrosis, endothelial vasoactive substance changes and endothelial dysfunction. Our purpose was to research the effect of an aqueous extract of Averrhoa carambola L. (AEA) on endothelial function in rats with ventricular remodelling induced by isoprenaline. Rats were subjected to injection of isoprenaline and administration of various drugs. Vasoactive substances were measured, and the ventricular remodelling index was detected by the weighing method. Immunohistochemical analysis, pathological examination, Western blot and Masson's trichrome staining were performed. After AEA administration, the levels of transforming growth factor-β (TGF-β), angiotensin II (AngII), inducible NO synthase (iNOS), endothelin-converting enzyme (ECE), and endothelin 1 (ET-1); the ventricular remodelling index; and the collagen volume fraction were decreased, while the levels of total NO synthase (tNOS) and endothelial NO synthase (eNOS) were increased. The pathological examination results showed that apoptosis, fibrosis, necrosis and inflammatory infiltration of myocardial tissue were attenuated by AEA treatment. AEA might alleviate ventricular remodelling in rats by maintaining the balance of vasoactive substances and the function of the vascular endothelium.


Value assessment of antimicrobials using the STEDI framework - How steady is the outcome?
Antimicrobial resistance (AMR) is one of the major threats to global population health, and the antimicrobial market requires substantial reimbursement reform and/or significant financial incentives to function properly. To address these challenges, England piloted a new health technology evaluation process in conjunction with a new payment model in 2019. The value assessment was performed using a dedicated broader value framework for antibiotics for the first time. This so-called STEDI framework is an acronym based on the five value elements it covers (Spectrum, Transmission, Enablement, Diversity, and Insurance value). Learnings from the pilot show that there are important considerations when implementing this value framework: The STEDI value profile of an antibiotic strongly depends on the local context and is impacted by trade-offs between individual value elements. Decision makers should therefore act carefully when applying STEDI to avoid distorting the overall evaluation result. Considering the STEDI value profile of an antibiotic is an important part of its value assessment as it allows for distinguishing between higher- and lower-value products. However, given the complexities surrounding its value assessment, further research must be undertaken to improve the overall STEDI evaluation process.


Importance of serum basal tryptase levels in children with insect venom allergy.
The importance of serum basal tryptase (sBT) levels on patients with venom allergy is highlighted in recent adulthood studies. The aim of this study was to evaluate the sBT levels of venom-allergic children with varying severity of clinical reactions. We also aimed to document the association between sBT levels and severe systemic reactions (SR). Serum basal tryptase levels were estimated by UniCAP (Pharmacia & Upjohn, Uppsala, Sweden). Children who suffered from large local reaction (LLR) or SR after insect stings were included along with healthy control subjects without a history of any local or SR after insect stings. A total of 128 children (55 with SR, 18 with LLR, and 55 age and sex-matched control subjects) with a median age of 8.9 years (range 3.2-17.4) were enrolled. Severe SR was encountered in 24 (44%) patients with SRs. The median level of sBT in children with SRs (median, interquartile range) [4.2 μg/l (3.6-4.9)] was significantly higher than in children with LLRs [3.1 μg/l (2.5-4.0)] and healthy control subjects [2.9 μg/l (2.3-3.4)] (P < 0.001). Logistic regression analysis revealed sBT ≥ 4.8 μg/l as a significant risk factor for severe SR (5.7 [1.5-21.4]; P = 0.01) in children with venom allergy. Our results indicate that sBT levels are associated with a higher risk of severe SR in children with insect venom hypersensitivity. Determination of sBT levels may help clinicians to identify patients under risk of severe SRs and optimal and timely use of therapeutic interventions in children with venom allergy.


[Corynebacterium riegelii as an unusual species separated from female patient with tuberculosis.]
The article describes a clinical case of tuberculosis of of urinary tracts of a female patient with massive discharge of strain Corynebacterium riegelii from urogenital tract. The data of clinical laboratory examination of female patient are analyzed. The characteristics of isolated strain C. riegelii (high degree of isolation rate (106 and higher) expressed urease activity (decomposition of urease during 5 min), presence of hemolytic activity) with consideration for estimation of its antibiotics sensitivity. The etiologic significance of isolated species C. riegelii in development of infection process in urogenital tract is demonstrated.


Temperature data during steam pressure filtration in combination with a water insoluble pore liquid.
Data presented in this article focus on the application of steam pressure filtration in combination with a water insoluble pore liquid. The article describes measured temperature profiles during steam pressure filtration within a filter cake. This article is co-submitted to the article 'Steam Pressure Filtration in Combination with a Water Insoluble Pore Liquid' [1] (DOI: 10.1016/j.ces.2020.115782) where the occurring phenomena as well as the interpretation of temperature profiles during steam pressure filtration are explained in detail. The article expands the shown data to other material systems.


Predictive value of C-reactive protein at different cutoff levels in acute appendicitis.
Determining the different cutoff values of C-reactive protein (CRP) on the basis of how long the patient's symptoms were present can be used to early predict acute appendicitis. We analyzed retrospectively from 2001 to 2004 the hospital records of 568 patients who underwent appendectomies for suspected appendicitis. Receiver operating characteristic analysis has shown that CRP measurement can increase the diagnostic accuracy in acute appendicitis. The cutoff values of CRP concentration taken as the first, second, and third days after onset of symptoms that distinguish acute appendicitis from other acute abdominal diseases were 1.5, 4.0, and 10.5 mg/dL, respectively; the values that distinguish perforated appendicitis from other acute abdominal diseases were 3.3 mg/dL (first day), 8.5 mg/dL (second day), and 12.0 mg/dL (third day). The different cutoff values of CRP concentration may serve as a useful predictive parameter in the early diagnosis of acute appendicitis on the first 3 days after the onset of symptoms.


Adjusted leptin level (ALL) is a predictor for hyperemesis gravidarum.
To compare the maternal serum leptin level according to the gestational week and to assess the relationship between the adjusted serum leptin level and thyroid hormones. In order to obtain this objective a new parameter is developed: adjusted leptin level (ALL: maternal serum leptin level/gestational week). A prospective study was carried out at the early prenatal care unit, SSK Ankara Women's Health and Teaching Hospital. Fifty-four women with hyperemesis gravidarum (HG) and 42 pregnant women without HG as a control group were included to the study. The groups were compared for age, parity, body mass index, fasting serum TSH, free T3, free T4 and leptin levels. A new parameter; ALL was also calculated in each case. Gestational age and body mass index (BMI) were found significantly lower in the HG group than in the control group (p=0.001). ALL was significantly high in the HG group (p=0.009). Serum TSH, free T3, free T4 levels were significantly different in the HG group than in the control group (p=0.003, 0.013, 0.012, respectively). A significant positive correlation was found between ALL and BMI in the HG group (r=0.449 p=0.001). The maternal leptin level was also positively correlated with BMI in the HG group (r=0.313 p=0.025). Etiology of the hyperemesis gravidarum is multifactorial. However we can postulate the adjusted leptin level is a good predictor for hyperemesis gravidarum.


Evaluation of the human serum albumin column as a discovery screening tool for plasma protein binding.
A total of 69 compounds with a variety of chemical structures were assayed using a human serum albumin column in combination with UV and mass spectrometric detection. A moderate correlation, R(2)=0.661, between the plasma protein binding, determined by traditional techniques of equilibrium dialysis or ultrafiltration, and chromatographic retention factor (k'/k'+1) was observed. Disparity between the regression line and numerous samples was observed across the entire range of plasma protein binding. Attempts to discriminate between compounds from the data set to achieve better correlation based physico-chemical properties were unsuccessful. Good agreement was observed for retention times obtained with UV detection with mobile phase containing phosphate buffer and mass spectrometric detection with mobile phase containing acetate buffer. Essentially identical data were obtained for compounds analyzed in singlet or cassette for minimally or highly bound (>90% bound) compounds. Analysis of cassettes containing compounds with plasma protein binding greater than 90% did not cause column overload, even at analyte concentrations up to 100 microg/ml. Diverse results were obtained when chromatographic retention was used to rank order various classes of compounds. Better correlation with ordering from known binding was obtained when a compound class contained a wide range of protein binding, in contrast to when compounds within a given class were all highly bound.


[Isolation and structure determination of two peptides occurring in human seminal plasma].
Two previously unknown peptides with a high amount of polar amino acids were isolated from human seminal plasma by a combination of dialysis, gel filtration, ion-exchange chromatography, and RP-HPLC. Their structures were determined by gas-phase sequencing simultaneously considering the different peak intensities.


Attractive Dispersion Interactions Versus Steric Repulsion of tert-Butyl groups in the Crystal Packing of a D3h -Symmetric Tris(quinoxalinophenanthrophenazine).
The crystalline packing of a π-extended D3h -symmetric triptycene reveals a particular π stacking motif with an almost-eclipsed arrangement of adjacent π planes despite the steric repulsion of tert-butyl substituents. Four model compounds were analyzed by using single-crystal X-ray diffraction and theoretical calculations to study the influence of dispersion interactions of molecular parts and understand the relationship between the molecular structure and this unique packing motif.


Integrative genomic meta-analysis reveals novel molecular insights into cystic fibrosis and ΔF508-CFTR rescue.
Cystic fibrosis (CF), caused by mutations to CFTR, leads to severe and progressive lung disease. The most common mutant, ΔF508-CFTR, undergoes proteasomal degradation, extinguishing its anion channel function. Numerous in vitro interventions have been identified to partially rescue ΔF508-CFTR function yet remain poorly understood. Improved understanding of both the altered state of CF cells and the mechanisms of existing rescue strategies could reveal novel therapeutic strategies. Toward this aim, we measured transcriptional profiles of established temperature, genetic, and chemical interventions that rescue ΔF508-CFTR and also re-analyzed public datasets characterizing transcription in human CF vs. non-CF samples from airway and whole blood. Meta-analysis yielded a core disease signature and two core rescue signatures. To interpret these through the lens of prior knowledge, we compiled a "CFTR Gene Set Library" from literature. The core disease signature revealed remarkably strong connections to genes with established effects on CFTR trafficking and function and suggested novel roles of EGR1 and SGK1 in the disease state. Our data also revealed an unexpected mechanistic link between several genetic rescue interventions and the unfolded protein response. Finally, we found that C18, an analog of the CFTR corrector compound Lumacaftor, induces almost no transcriptional perturbation despite its rescue activity.


Obstructive sleep apnea prevents the expected difference in craniofacial growth of boys and girls.
It was to compare cephalometric measures of mouth-breather boys and girls and with the cephalometric pattern observed in obstructive sleep apnea syndrome (OSAS) patients. Craniofacial measurements of lateral cephalometric radiographs obtained from 144 children aged 7-14 years were compared between boys and girls, and both were compared to cephalometric pattern of OSAS patients. Mouth-breather boys and girls had no gender differences regarding to craniofacial morphology while nose-breather boys and girls showed those expected differences. Nose-breather boys presented a more retruded mandible and proinclined upper incisor when compared to nose-breather girls, but mouth-breather boys and girls had no differences. The measure NS.GoGn was the only variable with an interaction with gender and breathing. There were no cephalometric difference in mouth breather-boys and girls related to normal growth, suggesting that oral breathing make the same craniofacial morphology and both have craniofacial morphology close to that of OSAS patients.


Analysis of rat liver Golgi apparatus proteins and apolipoproteins in acute ethanol intoxication.
Protein patterns of rat liver secretive (F1, F2) and formative (F3) Golgi fractions and their corresponding VLDLs content have been analyzed by disc-electrophoresis in normal conditions and after acute ethanol intoxication and compared with serum proteins and isolated lipoproteins. The major bands of the normal nascent VLDLs isolated from both formative and secretive Golgi fractions migrate in the same area of serum apo-C and apo-ARP apolipoproteins; in the same density interval of VLDLs another major lipoprotein band, called U, may be separated showing a slower electrophoretic mobility of apo-ARP. After ethanol intoxication apo-C bands seem to be concentrated in secretive and decreased in formative fractions, while apo-ARP and U bands show an overlapping in both Golgi fractions. The possible mechanisms of these behaviours are discussed.


A potent PGK1 antagonist reveals PGK1 regulates the production of IL-1β and IL-6.
Glycolytic metabolism enzymes have been implicated in the immunometabolism field through changes in metabolic status. PGK1 is a catalytic enzyme in the glycolytic pathway. Here, we set up a high-throughput screen platform to identify PGK1 inhibitors. DC-PGKI is an ATP-competitive inhibitor of PGK1 with an affinity of K d = 99.08 nmol/L. DC-PGKI stabilizes PGK1 in vitro and in vivo, and suppresses both glycolytic activity and the kinase function of PGK1. In addition, DC-PGKI unveils that PGK1 regulates production of IL-1β and IL-6 in LPS-stimulated macrophages. Mechanistically, inhibition of PGK1 with DC-PGKI results in NRF2 (nuclear factor-erythroid factor 2-related factor 2, NFE2L2) accumulation, then NRF2 translocates to the nucleus and binds to the proximity region of Il-1β and Il-6 genes, and inhibits LPS-induced expression of these genes. DC-PGKI ameliorates colitis in the dextran sulfate sodium (DSS)-induced colitis mouse model. These data support PGK1 as a regulator of macrophages and suggest potential utility of PGK1 inhibitors in the treatment of inflammatory bowel disease.


[Inhibitory effect of thalidomide combined with interferon on the proliferation of Kasumi-1 cells].
To explore the inhibitory effect of thalidomide combined with interferon (IFN) on the human acute myeloid leukemia cell line Kasumi- 1 and its mechanism. The inhibitiory effect of Kasumi- 1 cells by thalidomide, interferon or combination was detected by CCK- 8 method, the apoptosis by flow cytometry, the expression of apoptosis related proteins by Western blot, vascular endothelial growth factor (VEGF) concentration in culture supernatant by ELISA. Thalidomide inhibited the proliferation of Kasumi- 1 in a dose- dependent manner from 50 μg/ml to 500 μg/ml with an IC₅₀ of (451.13 ± 6.92)μg/ml at 24 h and (362.50 ± 14.52)μg/ml at 48 h. IFN also demonstrated the inhibitory capacity in a dose-dependent manner from 500 U/ml to 5 000 U/ml, with an IC₅₀ of (2 209 ± 127) U/ml at 24 h and (1 393±63) U/ml at 48 h. The apoptosis rates of Kasumi-1 cells treated with thalidomide 350 μg/ml or IFN 1 400 U/ml for 48 h were (14.68 ± 2.61) % and (21.71 ± 0.71)%, respectively, significantly higher than control group (P<0.01). In combination group the inhibition and the apoptosis rate were (88.50 ± 2.40) % and (41.95 ± 3.41)%, significantly higher than control and each single agent group (P<0.01). The VEGF concentrations of combination group [(94.61 ± 5.46) ng/L decreased significantly, as compared to thalidomide group [(141.11 ± 3.70) ng/L and IFN group [(119.90 ± 2.00) ng/L (P < 0.05). Western blot analysis showed Bcl-2 expression of Kasumi-1 cells decreased, while p-P38, Bax, cytochrome C, cleaved-Caspase-3, 8, 9 increased after treated with thalidomide 350 μg/ml or IFN 1 400 U/ml for 48 h. When treated with the combination agents, the expression of Bcl-2 further decreased and p-P38, Bax, cytochrome C, cleaved-Caspase-3, 8, 9 further increased as compared with each single agent (P < 0.05). Thalidomide and IFN could synergistically inhibit the proliferation of Kasumi-1 cells probably through inducing apoptosis via the mitochondrial pathway, death receptor pathway and P38 MAPK pathway, as well as inhibiting VEGF secretion.


Caffeic acid combined with autoclaved Leishmania major boosted the protection of infected BALB/c mice by enhancing IgG2 production, IFN-γ/TGF-β and iNO synthase/arginase1 ratios, and the death of infected phagocytes.
Immunization with killed Leishmania promastigotes without adjuvant was considered as safe, but gave variable rates of protection. Taking advantage of the immuno-modulatory effect of caffeic acid (CA), a natural polyphenolic antioxidant, we investigated its potentiating effect in autoclaved Leishmania major (ALM)-induced protection of Leishmania major-infected BALB/c. First, BALB/c mice were sensitized for 6 weeks either with CA, or ALM alone or combined with caffeic acid (ALM-CA) or Freund's adjuvant (ALM-FA), and subsequently infected with L. major promastigotes. Second, to test the curative effect, CA was given daily for 5 weeks to susceptible mice, starting on week 4 post-infection. Sera, footpads and lymph nodes (LNs) were collected at week 9 post-infection and submitted to biochemical or histological analyses. Compared to the respective controls, our results showed that CA directly healed footpad lesions and reduced the hallmarks of cutaneous leishmaniasis including oxidative inflammation, parasite load, and phagocytes influx and infestation. In sensitized mice, the protection enhanced gradually from ALM-FA, CA, ALM to ALM-CA in parallel to decreased seric IgGt levels. In contrast to ALM-FA, the combined effect of ALM and CA increased specific isotype IgG2, and decreased IL-17 and MCP-1, and phagocyte influx, as attested by the concomitant reduction in myeloperoxidase (MPO) and α-naphthyl acetate esterase (ANAE) activities. ALM-CA shifted IFN-γ/TGF-β and iNO synthase/arginase1 (iNOS/Arg1) balances in a Th1 immune response that control efficiently cutaneous lesions and LNs hypertrophy, and reactivate the death of infected phagocytes. Therefore, CA combined with ALM synergizes with L. major antigens for priming innate cells, through early polarization to optimal Th1 response that leads to IFN-γ and iNOS-dependent leishmanicidal activity of neutrophils and macrophages.


Eyetracking and selective attention in category learning.
An eyetracking version of the classic Shepard, Hovland, and Jenkins (1961) experiment was conducted. Forty years of research has assumed that category learning often involves learning to selectively attend to only those stimulus dimensions useful for classification. We confirmed that participants learned to allocate their attention optimally. We also found that learners tend to fixate all stimulus dimensions early in learning. This result obtained despite evidence that participants were also testing one-dimensional rules during this period. Finally, the restriction of eye movements to only relevant dimensions tended to occur only after errors were largely (or completely) eliminated. We interpret these findings as consistent with multiple-systems theories of learning which maximize information input in order to maximize the number of learning modules involved, and which focus solely on relevant information only after one module has solved the learning problem.


Short-Term Clinical Evaluation of Enhanced Unilateral Open-Door Laminoplasty Using Titanium Mesh.
Observational, cross-sectional study. The purpose of this study was to evaluate the clinical efficacy and safety profile of open-door laminoplasty (ODL) using titanium mesh. The most appropriate surgical management of degenerative spine disorders, particularly ossification of the posterior longitudinal ligament, remains controversial and continues unabated in the neurosurgical community. However, recently, ODL has become popular among surgeons. Many modifications have been developed since Hirabayashi in 1983 to enhance the clinical efficacy and safety of this procedure. We reviewed the obtained data of patients with ossification of the posterior longitudinal ligament (OPLL) who underwent ODL using titanium mesh at Neurosurgery Subdivision, the Dr. Sardjito Hospital, Yogyakarta, Indonesia, during January 2013-December 2015. Motor improvement (Nurick score), axial neck pain (neck disability index [NDI]), and incidence of complications were pre- and postoperatively examined, and the clinical efficacy and safety of the procedure was evaluated. Data were analyzed using Wilcoxon signed-rank test and paired t -test. This study included five patients were included (i.e., four males and one female) with mean age of 47.00±9.68 years, where reassessments were done after mean follow up of 20.2±4.74 months. Mean duration of surgery was 165±12.55 minutes. Pre- and postoperative motor assessments were 3.4±1.52 and 2.0±1.22 (p =0.059), respectively. Axial neck pains using NDI were 23.60±7.44 and 12.40±6.50 (p =0.004) before and after operations, respectively. No infected postoperative scar was found, and only one patient had a left C5 motor palsy. The results of the present study suggest that enhanced ODL using titanium mesh has favorable outcome and safety profile, which can pave the way for a suitable treatment in patients with OPLL.


"En-bloc" vertebrectomy in the mobile lumbar spine.
Primary tumors of the vertebral bodies have previously been treated with total or subtotal excision in a piecemeal fashion (intralesional excision). Radiation therapy has been used to help control tumor growth. Recurrence rates with an intralesional, piecemeal removal of vertebral tumors have been unacceptably high. This study describes a method to excise a lumbar vertebra "en-bloc," and in the process, to perform a marginal (extralesional) resection of a primary tumor of the mobile lumbar spine that allows for a potential surgical cure. A combined posterior-anterior procedure allows for an extralesional, marginal resection of the tumor and the involved vertebra. All posterior bony elements, including the pedicles and the adjacent intervertebral discs, are removed via a posterior approach. An anterior, retroperitoneal approach is then used to remove the vertebral body/tumor as a single specimen. The nerve roots at the involved levels are spared and the spine is instrumented and fused both posteriorly and anteriorly. Three patients successfully had combined posterior-anterior resections of lumbar vertebral chordomas. No permanent neurological complications occurred. Overall morbidity of the procedure was acceptable. At 31-month follow-up, no tumor recurrence has been detected. "En-bloc" resection of a primary vertebral tumor of the lumbar spine is technically demanding, but potentially curative. The alternative approaches-intralesional excision, radiation therapy, or a combination-are unable to cure these tumors. Long-term, 10-year follow-up will be necessary to confirm whether this en-bloc approach provides a surgical cure.


PDGF-R inhibition induces glioblastoma cell differentiation via DUSP1/p38MAPK signalling.
Glioblastoma (GBM) is the most common and fatal primary brain tumour in adults. Considering that resistance to current therapies leads to limited response in patients, new therapeutic options are urgently needed. In recent years, differentiation therapy has been proposed as an alternative for GBM treatment, with the aim of bringing cancer cells into a post-mitotic/differentiated state, ultimately limiting tumour growth. As an integral component of cancer development and regulation of differentiation processes, kinases are potential targets of differentiation therapies. The present study describes how the screening of a panel of kinase inhibitors (KIs) identified PDGF-Rα/β inhibitor CP-673451 as a potential differentiation agent in GBM. We show that targeting PDGF-Rα/β with CP-673451 in vitro triggers outgrowth of neurite-like processes in GBM cell lines and GBM stem cells (GSCs), suggesting differentiation into neural-like cells, while reducing proliferation and invasion in 3D hyaluronic acid hydrogels. In addition, we report that treatment with CP-673451 improves the anti-tumour effects of temozolomide in vivo using a subcutaneous xenograft mouse model. RNA sequencing and follow-up proteomic analysis revealed that upregulation of phosphatase DUSP1 and consecutive downregulation of phosphorylated-p38MAPK can underlie the pro-differentiation effect of CP-673451 on GBM cells. Overall, the present study identifies a potential novel therapeutic option that could benefit GBM patients in the future, through differentiation of residual GSCs post-surgery, with the aim to limit recurrence and improve quality of life.


On-column refolding and purification of recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) expressed as inclusion body in Escherichia coli.
Recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) was produced in E. coli as an inclusion body. rHuIL-1ra was purified to Over 98% purity by anion exchange chromatography after on-column refolding. The optimized processes produced more than 2 g pure refolded rHuIL-1ra per 1 l culture, corresponding to a 44% recovery, without an intermediate dialysis step. Refolded rHuIL-1ra had full biological activity with the MTT assay. An intramolecular disulfide linkage in the oxidized recombinant protein was suggested by data from HPLC and non-reducing SDS-PAGE.


Land use, spatial scale, and stream systems: lessons from an agricultural region.
We synthesized nine studies that examined the influence of land use at different spatial scales in structuring biotic assemblages and stream channel characteristics in southeastern Minnesota streams. Recent studies have disagreed about the relative importance of catchment versus local characteristics in explaining variation in fish assemblages. Our synthesis indicates that both riparian- and catchment-scale land use explained significant variation in water quality, channel morphology, and fish distribution and density. Fish and macroinvertebrate assemblages can be positively affected by increasing the extent of perennial riparian and upland vegetation. Our synthesis is robust; more than 425 stream reaches were examined in an area that includes a portion of three ecoregions. Fishes ranged from coldwater to warmwater adapted. We suggest that efforts to rehabilitate stream system form and function over the long term should focus on increasing perennial vegetation in both riparian areas and uplands and on managing vegetation in large, contiguous blocks.


Bacterial cellulose production by Gluconacetobacter sp. PKY5 in a rotary biofilm contactor.
A rotary biofilm contactor (RBC) inoculated with Gluconacetobacter sp. RKY5 was used as a bioreactor for improved bacterial cellulose production. The optimal number of disk for bacterial cellulose production was found to be eight, at which bacterial cellulose and cell concentrations were 5.52 and 4.98 g/L. When the aeration rate was maintained at 1.25 vvm, bacterial cellulose and cell concentrations were maximized (5.67 and 5.25 g/L, respectively). The optimal rotation speed of impeller in RBC was 15 rpm. When the culture pH in RBC was not controlled during fermentation, the maximal amount of bacterial cellulose (5.53 g/L) and cells (4.91 g/L) was obtained. Under the optimized culture conditions, bacterial cellulose and cell concentrations in RBC reached to 6.17 and 5.58 g/L, respectively.


Comparing Medicaid Expenditures for Standard and Enhanced Therapeutic Foster Care.
Therapeutic foster care (TFC) is a service for children with high behavioral health needs that has shown promise to prevent entry into more restrictive and expensive care settings. The purpose of this study was to compare Medicaid expenditures associated with TFC with Medicaid expenditures associated with an enhanced higher-rate service called Intensive Alternative Family Treatment (IAFT). We conducted a secondary analysis of Medicaid claims in North Carolina among children entering care in 2018-2019. Using propensity score analysis with difference-in-difference estimation, we compared monthly Medicaid expenditures before and after initiating TFC and IAFT (N = 5472 person-months). Youth entering IAFT had higher expenditures prior to treatment than those entering TFC. Both standard TFC and IAFT were associated with a downward trend in expenditures following treatment initiation. Both TFC and IAFT reverse a trend of increasing Medicaid costs prior to care among children with high behavioral health needs.


How to structure an oncogenetics service for the public health system: Report of the implementation of the first service in Northeastern Brazil.
Identifying carriers of genetic mutations that increase the risk of developing cancer allows to adopt timely risk-reducing strategies. However, due to the elevated cost of genetic testing, few oncogenetics services are available in the Brazilian public health care system, especially in economically disadvantaged areas. To describe the implementation of an oncogenetics service for patients suspected of hereditary cancer syndromes (HBOC and HNPCC) at a philanthropic referral oncology hospital in Northeastern Brazil, funded by the Ministry of Health's National Oncology Care Support Program (PRONON). The service was implemented with the PDCA method (Plan, Do, Check and Act). During the first year of operation (starting in August 2018), 675 individuals were examined, of whom 272 patients and 98 family members were submitted to genetic testing. This included the collection of 338 DNA samples of which 300 were sequenced. The analysis identified 48 (17.1%) mutations for HBOC and 19 (6.8%) for HNPCC. In one year, the oncogenetics service was able to benefit over 300 families by generating advanced molecular data which may be used for tailoring cancer prevention and management.


XO/XY mosaicism and non-fluorescing Y chromosome in a male.
An adult male of short stature and with underdeveloped external genitalia is described, who carried out a number of sexual assaults on young women. He proved to have XO/XY mosaicism and a non-fluorescing Y chromosome. It was considered to be a terminal deletion on morphological grounds. It is suggested, on the evidence of the small number of XO/XY mosaics examined by appropriate staining methods, that an abnormal Y chromosome, whether terminally deleted or non-fluorescing owing to an altered chemical state, predisposes to anaphase lagging and non-disjunction. Of eleven reported cases of XO/XY mosaicism with a non-fluorescing Y chromosome, this is the fifth of male phenotype. The severe behaviour disturbance of early onset is considered to be probably causally associated with the chromosome anomaly.


Duplex ultrasonography of vertebral and subclavian arteries.
Duplex ultrasonography of the vertebral artery is an integral part of extracranial cerebrovascular screening and provides noninvasive diagnostic clues for subclavian or vertebral artery stenosis. This review describes technique for the ultrasound examination of the vertebral and subclavian arteries. Duplex images from various patients are used to illustrate a spectrum of hemodynamic changes that occur in the vertebral artery Doppler signal caused by subclavian artery stenosis and subclavian-vertebral steal.


Bis(oxotremorine) fumarate bis-(fumaric acid).
The title compound, bis-(oxotremorine) fumarate bis-(fumaric acid) {systematic name: 1-[4-(2-oxopyrrolidin-1-yl)but-2-yn-yl]pyrrolidinium (2E)-but-2-ene-di-o-ate bis-[(2E)-but-2-enedioic acid]}, 2C12H19N2O+·C4H2O4 2-·2C4H4O4, has a single oxotremorine monocation protonated at the pyrrolidine nitro-gen, one fumaric acid mol-ecule and half of a fumarate dianion in the asymmetric unit. The ions and fumaric acid mol-ecules are held together by N-H⋯O and O-H-⋯O hydrogen bonds in 40-membered rings with graph-set notation R 6 6(40). The fumarate ions join these rings into infinite chains along [001].


Effects of deacetylation degree of chitosan on the structure of aerogels.
Chitosan aerogels, obtained by (supercritical) CO2 drying of hydrogels, are novel adsorbents because of their large surface area and high porosity. Intrinsic properties of chitosan such as molecular weight (MW) and degree of deacetylation (DDA) had large impacts on the characteristics of chitosan aerogels. Although there are a few studies about the effects of solely DDA or MW on aerogel structure, none of them has focused on the mutual effects. The study aims to investigate the combined effects of MW and DDA of chitosan on aerogel properties. Hydrogels were produced in beads form by physical gelation of the chitosan solutions (2 % w/v in acetic acid of 1 %, v/v) in an alkaline environment (NaOH, 4 N). Supercritical CO2 dried aerogels were examined with respect to the bulk density, diameter as well as pore characteristics, and surface area by Barrett-Joyner-Halenda (BJH) and Brunauer-Emmett-Teller (BET) methods, respectively. Morphologies of aerogels were also examined by Scanning Electron Microscopy (SEM) images and structural changes of aerogels were observed by Fourier Transform Infrared (FTIR) Spectroscopy. Additional to BET-BJH analysis, proton relaxation dispersion was measured by Fast Field Cycling NMR (FFC-NMR) to determine the pore volume of the aerogels. Compact structures were obtained for higher MW chitosan and lower MW chitosans with higher DDA increasing the aerogel diameters. All types of aerogels obtained by different chitosan characteristics (MW and DDA) showed a porous structure and the highest DDA with the lowest MW caused the minimum bulk density with the highest water absorption rate. Although different N2 adsorption-desorption profiles were obtained in terms of pore volumes; all aerogels had Type IV isotherms with Type H1 hysteresis curve. FFC-NMR experiments showed that the coherence length values were associated with the pore volumes and FFC-NMR experiments were found to be meaningful as supportive experiments for the characterization of aerogels.


Evaluation of the Effect of NT-3 and Biodegradable Poly-L-lactic Acid Nanofiber Scaffolds on Differentiation of Rat Hair Follicle Stem Cells into Neural Cells In Vitro.
Recent improvement in neuroscience has led to new strategies in neural repair. Hair follicle stem cells are high promising source of accessible, active, and pluripotent adult stem cells. They have high affinity to differentiate to neurons. Aside from using cell-scaffold combinations for implantation, scaffolds can provide a suitable microenvironment for cell proliferation, migration, and differentiation. NT-3 is the most interesting neurotrophic factors being an important regulator of neural survival and differentiation. Since treatment duration in neural repair is very important, this study aims to evaluate the effect of NT-3 and poly-L-lactic acid (PLLA) on differentiation time of bulge stem cells of rat hair follicle to neural-like cells. HFSCs of rat whisker was isolated and cultured on PLLA and differentiated with 10 ng/mL NT-3. Biological features of cultured cells were evaluated with immunocytochemistry and flowcytometry methods by using CD34, nestin, and βІІІ-tubulin markers. For cell viability and morphological assessment, MTT assay and SEM were performed. Our results showed that bulge stem cells of hair follicle can express CD34 and Nestin before differentiation. By using NT-3 during differentiation process, the cells showed positive reaction to βІІІ-tubulin antibody. MTT results demonstrated that PLLA significantly increased cell viability. Finally, HFSCs adhesion was confirmed by SEM results. The results indicate that 10 ng/mL NT-3 and PLLA have significant effect on differentiation time of rat HFSCs to neural cells even in 10 days.


Cytapheresis (CAP) with leukocyte removal filter/bead column as one therapeutic option for inflammatory bowel disease.
Inflammatory bowel disease (IBD) like Crohn's disease and ulcerative colitis are chronic inflammatory disorders that affect the bowel. The disease is characterized by periods of clinical remission and relapse due to severe intestinal inflammation. Drug therapy of IBD is associated with unpleasant side effects. Further, efficacies of conventional drugs decrease with chronic use and this can represent a major difficulty in the long term management of IBD. However, in active IBD, leukocytes are elevated in the lesion they may be able to be a factor of IBD aggravation. Membrane filters column and leukocyte adsorbing beads have been developed which are direct blood perfusion systems for removing any desired level of leukocytes. Clinical studies with these two new models have shown good effects for active IBD. Clinical data suggest that leukocytapheresis might be an effective adjunct to therapy of IBD, to promote remission, taper conventional drug dosage and potentially should reduce the number of patients who require colectomy. The results may further understandings of the pathophysiology of IBD and this in turn should contribute to a more effective treatment of this disorder.


Diverse single-stranded DNA viruses identified in New Zealand (Aotearoa) South Island robin (Petroica australis) fecal samples.
The South Island robin (Petroica australis) is a small passerine bird endemic to New Zealand (Aotearoa). Although its population has declined recently and it is considered 'at risk,' little research has been done to identify viruses in this species. This study aimed to survey the diversity of single-stranded DNA viruses associated with South Island robins in a small, isolated population on Nukuwaiata Island. In total, 108 DNA viruses were identified from pooled fecal samples collected from 38 individual robins sampled. These viruses belong to the Circoviridae (n = 10), Genomoviridae (n = 12), and Microviridae (n = 73) families. A number of genomes that belong to the phylum Cressdnaviricota but are otherwise unclassified (n = 13) were also identified. These results greatly expand the known viral diversity associated with South Island robins, and we identify a novel group of viruses most closely related genomoviruses.


Receptors for endogenous and heterogenous hydroxamate siderophores in Staphylococcus aureus B 471.
In Staphylococcus aureus B47 grown in iron-restricted medium, six new, iron-regulated proteins occurred in cytoplasmic membrane. Protein of 14kDa has bound two complexes of iron: Fe(III)-staphylobactin and Fe(III)-acinetoferrin. Complexes of Fe(llI)-ferrichrome and Fe(III)-rhodotorulic acid were not bound to any of new membrane protein. Iron of Fe(III)-staphylobactin and Fe(IIl)-acinetoferrin complexes was transported into the cells.


Predictors of return to work ten months after primary breast cancer surgery.
The most common female cancer in Western countries is breast cancer and women diagnosed with this disease are often under 65 years old. With increasing prevalence of survivors it is important to shed light on problems facing these women after diagnosis and treatment. The aim of this study was to assess factors predicting return to work (RTW) in women with early-stage breast cancer. A cohort of 102 women aged 18-64 with early-stage breast cancer who had undergone curative primary surgery with or without systemic adjuvant therapy were followed for 10 months using data from questionnaires and medical files. Ten months after primary surgery, 59% of the women had returned to work while 41% were sick-listed part-time or full-time. After adjusting for age, health status, life satisfaction, vocational situation, and irradiation to the breast/chest wall and regional nodes, a multivariate logistic regression revealed the following factors as being negatively associated with RTW: a high-demand job (OR=0.1, 95% CI 0.0-0.8), axillary node dissection (OR=0.1, 95% CI 0.0-0.6), and treatment with chemotherapy (OR=0.1, 95% CI 0.0-0.7). Treatment factors and high demands at work play an important role in RTW for women with early-stage breast cancer.


Separation and quantitation of azimilide and its putative metabolites by capillary electrophoresis.
Reliable methods based on capillary electrophoresis (CE) have been developed for the separation and quantitation of azimilide, an antiarrhythmic drug under development at Procter & Gamble Pharmaceuticals (P&GP). Both capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) were employed in the separation of azimilide from its impurities, degradants and/or metabolites. Separation of azimilide from NE-11178, F-410, F-1054 and F-1292 was obtained by MECC at pH 9 with 50 mM sodium dodecyl sulfate (SDS). The separation of azimilide and NE-10171, a key metabolite of azimilide, was difficult because their structures differ by only a single methyl group. The best separation was achieved under acidic pH conditions with cetyltriethyl ammonium chloride (CTAC) additive in the buffer. All of the CE separations were completed within a substantially shorter time and with better resolution than the corresponding high-performance liquid chromatography (HPLC) separations. Quantitation was done with azimilide and NE-10171. Calibration curves ranging from 10 to 1000 microg/ml were obtained with R2 greater than 0.997 for both azimilide and NE-10171. The back-calculated concentrations of the calibration standards and the recoveries of the quality control (QC) samples were within the acceptance range currently used for HPLC methods. These results demonstrated the viability of CE as an alternative technique for drug metabolism studies in support of pharmaceutical development.


Effect of the FreeStyle Libre™ flash glucose monitoring system on glycemic control in individuals with type 2 diabetes treated with basal-bolus insulin therapy: An open label, prospective, multicenter trial in Japan.
We investigated the effect of FreeStyle LibreTM on glycemic control in Japanese type 2 diabetes patients treated with basal-bolus insulin therapy. This prospective, 90-day single-arm study enrolled 94 adults with type 2 diabetes treated with insulin. A 14-day masked baseline phase was followed by an 11-week treatment phase during which participants used the device to monitor glucose levels. The primary end-point was time spent in hypoglycemia (<70 mg/dL) for baseline versus study end (days 76-90). Secondary end-points included other measures of glycemic control, along with patient satisfaction using the Japanese Diabetes Treatment and Satisfaction Questionnaire. Time spent in hypoglycemia was low at baseline (0.51 ± 0.93 h/day) and did not significantly decrease at study end (0.47 ± 0.63 h/day, P = 0.6354). Time in range, time in hyperglycemia and estimated A1c all improved versus baseline (by +1.7 ± 3.0 h/day, -1.6 ± .4 h/day and -0.4 ± 0.8%, respectively, P < 0.0001 in each). Finger stick tests fell from 2.9 ± 1.3 to 1.9 ± 1.4/day, and mean scanning frequency during the intervention phase was 11.3/day. The mean treatment satisfaction score increased by 11.8 ± 5.3 (P < 0.0001). Two severe hypoglycemia-related adverse events were reported; one of which was possibly related to the device. Three participants reported mild device-related skin trauma, site discomfort or subcutaneous bleeding. Use of FreeStyle Libre by Japanese type 2 patients diabetes treated with basal-bolus insulin therapy showed a low baseline of hypoglycemia, and enabled improved glycemic control and treatment satisfaction.


Validation of the revised diagnostic criteria for primary plasma cell leukemia by the Korean Multiple Myeloma Working Party.
The International Myeloma Working Group has recently revised the diagnostic criteria for primary plasma cell leukemia (PCL) to circulating plasma cells (CPCs) ≥ 5% in a peripheral blood smear. The present study validated new criteria in patients with multiple myeloma or PCL diagnosed using the previous diagnostic criteria, who were administered immunomodulatory drugs or proteasome inhibitors as induction therapy. We analyzed the medical records of 1357 patients from eight hospitals in South Korea. The median age of the all patients was 64 years, and 187 (13.8%) had CPCs at diagnosis. Only 79 (5.8%) of the patients had ≥ 5% CPCs. The median overall survival (OS) of patients with CPCs ≥ 5% and ≥ 20% was similar, but had significantly inferior median progression-free survival (PFS) and median OS than those with CPCs < 5% (13.1 vs. 21.5 months, P < 0.001, and 21.5 vs. 60.9 months, P < 0.001, respectively). Primary PCL diagnosed using the revised criteria presented with higher total calcium levels and serum creatinine levels, lower platelet counts and frequent organomegaly and plasmacytoma at diagnosis. Univariate and multivariate analyses demonstrated that the presence of plasmacytoma and elevated serum β2-microglobulin were significantly associated with OS in primary PCL. In conclusion, the revised criterion of CPCs ≥ 5% in a peripheral blood smear is appropriate for PCL diagnosis.


Pragmatic Clinical Studies: An Emerging Clinical Research Discipline for Improving Evidence-Based Practice of Cardiovascular Diseases in Asia.
Pragmatic clinical studies, an emerging clinical research discipline, include a wide range of studies that are largely embedded with routine clinical practice and aim to evaluate the comparative effectiveness and safety of different clinical intervention strategies. Increased availability and quality of electronic medical/health records drives the development of pragmatic clinical studies. In this review, we describe evolution of the conceptual framework of pragmatic clinical studies and share perspectives on the importance of pragmatic clinical studies in evidence-based practice for cardiovascular diseases, as a complement to conventional randomized controlled trials. We also highlight specific needs of pragmatic clinical studies in improving evidence-based practice for cardiovascular disease in Asian countries. The main challenges of pragmatic clinical studies are discussed briefly in this review.


Polymer-coated pH-responsive high-density lipoproteins.
Intracellular drug delivery by nanoparticles is often hampered by their endosomal entrapment followed by their degradation in the lysosomal compartment and/or exocytosis. Here, we show that internalization and endosomal escape of cargoes in a cationized natural nanocarrier, high-density lipoprotein (HDL), can be controlled in a pH-dependent manner through stable complexation with a membranolytic anionic block polymer. A genetically and chemically cationized form of HDL (catHDL) is prepared for the first time by both genetic fusion with YGRKKRRQRRR peptide and incorporation of 1,2-dioleoyloxy-3-(trimethylammonium)propane. Upon addition of poly(ethylene glycol)-block-poly(propyl methacrylate-co-methacrylic acid) (PA), catHDL yields inhibition of internalization at neutral pH and its subsequent recovery at mildly acidic pH. catHDL forms a stable discoidal-shape complex with PA (catHDL/PA) (ca. 50 nm in diameter), even in the presence of serum. Significant enhancement of endosomal escape of a catHDL component is observed after a 1-h treatment of human cancer cells with catHDL/PA. Doxorubicin and curcumin, fluorescent anti-cancer drugs, encapsulated into catHDL/PA are also translocated outside of endosomes, compared with that into catHDL, and their cytotoxicities are enhanced inside the cells. These data suggest that catHDL/PA may have a potential benefit to improve the cellular delivery and endosomal escape of therapeutics under mildly acidic conditions such as in tumor tissues.


Allogeneic hemopoietic stem cell transplantation in solid tumors.
The existence of a graft versus tumor (GVT) effect of donor-derived T cells after allogeneic hematopoietic stem cell transplantation is well established as a critical component for the success of the procedure in several hematologic malignancies. Although it has been suggested that a GVT effect might also be generated in patients affected by refractory solid tumors, the morbidity of conventional allogeneic hematopoietic stem cell transplantation has limited its investigation in these diseases. Recently introduced allogeneic nonmyeloablative regimens have greatly decreased morbidity and mortality related to transplants which retain a powerful GVT. On this basis, it has become possible to explore the existence of alloreactivity toward solid tumors. The present article reviews the early clinical results of this novel immunotherapeutic approach for solid tumors.


Intraneuronal cylindrical particles in Alzheimer's disease.
In this report we describe an unusual form of intraneuronal cylindrical particles (CP) in the brain of a 69-year-old man with typical Alzheimer's disease (AD). A large number of CP were seen in cortical neurons in the frontal and parietal lobes. The CP were always seen within the cisternae of endoplasmic reticulum, had a 25- to 30-nm-thick trilaminar wall and measured 80-95 nm in cross-sectional diameter. In an occasional neuron, the CP were seen next to filamentous constituents of neurofibrillary tangles. The remaining neurons containing CP appeared normal. A similar finding has been reported in another case of AD and also in two cases with other conditions, thus indicating that the presence of CP is not limited to AD. Morphologically similar intracisternal CP have been observed in the macrophages in certain strain of mice following implantation of dibenzanthracene. The CP in human brains and experimental animals show a remarkable resemblance to rhabdovirus and certain murine endogenous virus particles. The nature of these intraneuronal CP in human brains and their significance, if any, remain undetermined.


Topological bootstrap: Fractionalization from Kondo coupling.
Topologically ordered phases of matter can host fractionalized excitations known as "anyons," which obey neither Bose nor Fermi statistics. Despite forming the basis for topological quantum computation, experimental access to these exotic phases has been very limited. We present a new route toward realizing fractionalized topological phases by literally building on unfractionalized phases, which are much more easily realized experimentally. Our approach involves a Kondo lattice model in which a gapped electronic system of noninteracting fermions is coupled to local moments via the exchange interaction. Using general entanglement-based arguments and explicit lattice models, we show that gapped spin liquids can be induced in the spin system, and we demonstrate the power of this "topological bootstrap" by realizing chiral and Z2 spin liquids. This technique enables the realization of many long sought-after fractionalized phases of matter.


Efficacy of adjunctive measures in the non-surgical treatment of peri-implantitis: A systematic review.
The aim of this systematic review was to evaluate the efficacy of patient-performed or administered adjunctive measures to non-surgical peri-implantitis therapy in terms of probing depth (PD) and/or bleeding on probing (BoP) reductions. Randomized and controlled clinical trials with at least 6 months of follow-up were searched in three databases. Secondary outcomes included implant loss, disease resolution, recurrence of peri-implantitis, need of re-treatment, changes in marginal bone levels, patient-reported outcomes and adverse effects. Of 567 titles, 10 publications, reporting 9 investigations, were included. Three types of adjunctive measures were found (local/systemic antimicrobials and probiotics). Four studies evaluated the effects of local antimicrobials (i.e., minocycline microspheres, chlorhexidine chips or a metronidazole + amoxicillin gel), three studies evaluated systemic antimicrobials (either amoxicillin + metronidazole or metronidazole alone) and two studies evaluated probiotics (Lactobacillus reuteri strains). The addition of local antimicrobials led to modest improvements in PD reduction. Systemic antimicrobials showed significantly greater reductions in PD and BoP, especially at initially deep sites (PD > 6 mm). Due to the large heterogeneity among included studies, no meta-analyses were performed. Different adjunctive measures in the non-surgical treatment of peri-implantitis have different impact in terms of PD and BoP reductions. Improved PD reductions result after the use of systemic antimicrobials, and to a lesser extent, after the use of local antimicrobials.


Translation of mRNA from rat kidney following acute exposure to lead.
1. Two days after the administration of Pb2+ (0.54 mg/100 body wt) to rats, the kidneys were removed and homogenate fractions obtained. 2. The machinery for peptide synthesis exhibited increased activity in the kidney preparations from lead-treated rats. 3. The poly(A)+ RNA was obtained from the polysome fraction and translated in a mRNA-dependent reticulocyte lysate system. 4. The translatability of the poly(A)+ RNA was similar using the preparations obtained from control and lead-treated rats except for proteins of approx molecular weight of 50,000 which contained increased amounts [14C]leucine in the fractions derived from lead-treated rats. 5. The results are compared with those after acute Cd2+ toxicity where poly(A)+ RNA exhibited increased translatability for low molecular weight metallothioneins.


Draft Genome Sequence of a Putative Raw Milk-Associated Probiotic Bacterium, Pediococcus acidilactici ISO17.
Pediococcus acidilactici is a beneficial lactic acid bacterium frequently studied for its probiotic potential. Here, we present a 2.32-Mb draft genome sequence of strain ISO17, with a G+C content of 42%. The genome was predicted to harbor 4 rRNA genes, 47 tRNA genes, and 2,297 protein-coding sequences.


Maternal depressive symptoms, rumination, and child emotion regulation.
Children of depressed mothers are at risk for maladaptive emotion regulation. This study examined a model of maternal rumination that links maternal depressive symptoms to child emotion regulation. A sample of 126 mother-child dyads (65 girls) participated in the current study, at three assessment points when children were age three, four, and five. At all assessment points, mothers reported their depressive symptoms and ruminative response style. Child emotion regulation was assessed observationally from two laboratory tasks, which were designed to elicit anger and sadness. Elevated maternal depressive symptoms were associated with higher concurrent rumination. Maternal rumination at child age four predicted that more child focus on distress during sadness-eliciting tasks and less child active distraction during anger-eliciting tasks 1 year later. Additionally, maternal rumination at child age three and four predicted less child passive behaviors in anger-eliciting tasks prospectively. Findings suggest that maternal rumination is predictive of using maladaptive regulatory strategies among children of depressed mothers. This study has significant implications for intervention programs targeting depressed mothers and their children.


Liver Steatosis: A Marker of Metabolic Risk in Children.
Obesity is one of the greatest health challenges affecting children of all ages and ethnicities. Almost 19% of children and adolescents worldwide are overweight or obese, with an upward trend in the last decades. These reports imply an increased risk of fat accumulation in hepatic cells leading to a series of histological hepatic damages gathered under the acronym NAFLD (Non-Alcoholic Fatty Liver Disease). Due to the complex dynamics underlying this condition, it has been recently renamed as 'Metabolic Dysfunction Associated Fatty Liver Disease (MAFLD)', supporting the hypothesis that hepatic steatosis is a key component of the large group of clinical and laboratory abnormalities of Metabolic Syndrome (MetS). This review aims to share the latest scientific knowledge on MAFLD in children in an attempt to offer novel insights into the complex dynamics underlying this condition, focusing on the novel molecular aspects. Although there is still no treatment with a proven efficacy for this condition, starting from the molecular basis of the disease, MAFLD's therapeutic landscape is rapidly expanding, and different medications seem to act as modifiers of liver steatosis, inflammation, and fibrosis.


A low prevalence of MYH7/MYBPC3 mutations among familial hypertrophic cardiomyopathy patients in India.
Familial Hypertrophic Cardiomyopathy (FHC) is an autosomal dominant disorder affecting the cardiac muscle and exhibits varied clinical symptoms because of genetic heterogeneity. Several disease causing genes have been identified and most code for sarcomere proteins. In the current study, we have carried out clinical and molecular analysis of FHC patients from India. FHC was detected using echocardiography and by analysis of clinical symptoms and family history. Disease causing mutations in the β-cardiac myosin heavy chain (MYH7) and Myosin binding protein C3 (MYBPC3) genes were identified using Polymerase Chain Reaction-Deoxyribose Nucleic Acid (PCR-DNA) sequencing. Of the 55 patient samples screened, mutations were detected in only nineteen in the two genes; MYBPC3 mutations were identified in 12 patients while MYH7 mutations were identified in five, two patients exhibited double heterozygosity. All four MYH7 mutations were missense mutations, whereas only 3/9 MYPBC3 mutations were missense mutations. Four novel mutations in MYBPC3 viz. c.456delC, c.2128G>A (p.E710K), c.3641G>A (p.W1214X), and c.3656T>C (p.L1219P) and one in MYH7 viz. c.965C>T (p.S322F) were identified. A majority of missense mutations affected conserved amino acid residues and were predicted to alter the structure of the corresponding mutant proteins. The study has revealed a greater frequency of occurrence of MYBPC3 mutations when compared to MYH7 mutations.


Molecular cloning, characterization and immunological response analysis of Toll-like receptor 21 (TLR21) gene in grass carp, Ctenopharyngodon idella.
TLR21, a non-mammalian Toll-like receptor, has been recently identified in fishes, frogs and birds. In the present study, the full-length cDNA sequence of TLR21 (CiTLR21) from Ctenopharyngodon idella has been isolated and characterized. The CiTLR21 full-length cDNA sequence consists of 3578bp, with an open reading frame (ORF) of 2958bp encoding 985 amino acid residues. The putative CiTLR21 protein contains a signal peptide sequence, 17 leucine-rich (LRR) motifs, a transmembrane region and a Toll/interleukin-1 receptor (TIR) domain. The CiTLR21 gene is expressed in a wide range of tissues with the highest expression in skin. Upon induction by Aquareovirus, CiTLR21 expression is significantly down-regulated in liver and spleen, whereas is significantly up-regulated in liver and spleen after Aeromonas hydrophila infection. These results suggest that CiTLR21 plays an important role in Aquareovirus and A. hydrophila-related diseases. This work may provide the basis for further investigations into the immune system of grass carp and other teleost fishes.


Evaluation of changes in egg yolk lipids during storage based on lipidomics through UPLC-MS/MS.
A comprehensive UPLC-MS/MS-based lipidomics analysis of egg yolk at different storage periods was performed. Through the analysis, 836 lipid species from 27 subclasses were detected, including 159 significantly different lipid species. Glycerolipids (GL) and glycerophospholipids (GP) were the most abundant lipids, followed by fatty acyls (FA), sphingolipids (SP), sterol lipids (ST), and prenol lipids (PR). PCA and OPLS-DA showed that egg yolk lipids changed significantly with prolonged storage. Moreover, glycerophospholipid metabolism and glycerolipid metabolism were key metabolic pathways, as phosphatidic acid (PA), lysophosphatidic acid (LPA), and diglyceride (DG) were key lipid metabolites in the related metabolic pathways of egg yolk during storage. Furthermore, egg yolk quality changed the most when stored up to 21 days. Our study establishes a foundation for the quality control and the lipid oxidation mechanism of eggs during storage.


Electrophoretic characterizations of cross-linked fibrinogen derivatives in blood and vascular tissue by zonal immobilization on glyoxyl agarose.
Direct immunoprobing of electrophoregrams of plasma and intimal protein on glyoxyl agarose and composite-gels with polyacrylamide have uncovered novel modes of cross-linking of fibrinogen that differ from those previously characterized. These modes of cross-linking involve the A alpha-chains of fibrinogen and hybrid cross-linking of alpha- and gamma-chains.


Coronary angiography- or fractional flow reserve-guided complete revascularization in multivessel disease STEMI: A Bayesian hierarchical network meta-analysis.
To identify the best strategy to achieve complete revascularization (CR) in patients with ST-elevation myocardial infarction (STEMI) and multi-vessel disease (MVD). We systematically reviewed the literature for randomized controlled trials (RCTs) comparing IRA-only PCI and CR guided by angiography or fractional flow reserve (FFR) in MVD-STEMI. Both frequentist (classical) and Bayesian network meta-analysis were performed, including a comparative hierarchy estimation of the probability to reduce the primary composite endpoint of all-cause death and new myocardial infarction (MI). We identified 11 RCTs, including 8193 STEMI patients. Compared with IRA-only strategy, CR significantly reduced the primary endpoint (OR: 0.73; 95%CI0.55-0.97). We observed non-significant difference between angiography and FFR guidance in reducing the primary endpoint (OR: 0.73, 95% CI 0.35-1.57). The Bayesian probability analysis ranked angio-guided CR as the best intervention yielding lowest risk of all-cause death or new MI (SUCRA92%). In patients with MVD-STEMI, CR is associated with a reduction in all-cause mortality and new MI compared with IRA-only PCI. Angio-guided CR is associated with the lowest risk of all-cause death or new MI, therefore the role of FFR-guidance in this setting is questionable. Both frequentist and Bayesian network meta-analysis were performed to compare infarct-related artery (IRA)-only percutaneous coronary intervention (PCI) and complete revascularization (CR) guided by angiography or fractional flow reserve (FFR) in multivessel disease (MVD) and acute ST-elevation myocardial infarction (STEMI). Eleven randomized controlled trials were identified, including 8193 STEMI patients. Compared with IRA-only strategy, CR significantly reduced the incidence of the composite endpoint of all-cause death and new myocardial infarction without significant difference in angio-guided and FFR-guided CR. The Bayesian probability analysis ranked angio-guided CR as the best intervention yielding lowest risk of the composite endpoint and, therefore the role of FFR-guidance in this setting is questionable.


Deregulation of purinergic ectoenzyme activity in head and neck cancer promotes immunosuppression.
Head and neck cancer (HNC) comprises a spectrum of neoplasms that affect the upper aerodigestive tract and are the sixth most common cancers worldwide. Individuals with HNC exhibit various symptoms and metabolic changes, including immune alterations and alterations of the purinergic pathway, which may signal worse outcomes. Therefore, the purpose of this research was to measure the activity of purinergic ectoenzymes and interleukins in patients with HNC, oral cavity cancer, and larynx cancer. We recruited 32 patients and 33 healthy control subjects and performed the laboratory analyses. We identified dysregulation in the purinergic signaling pathway characterized by an increase in adenosine triphosphate (ATP) and adenosine monophosphate (AMP) hydrolysis and a decrease in the deamination of adenosine to inosine in these cancers (p < 0.05). These alterations were likely caused by increased activity of the ectoenzymes E-NTPDase and ecto-5'-nucleotidase and reduced adenosine deaminase activity. This dysregulation was associated with immune alterations, increased levels of IL-10, and decreased myeloperoxidase activity (p < 0.05), suggesting immunosuppression in these patients and suggesting possible accumulation of adenosine in the extracellular environment. Adenosine is a potent immunosuppressive molecule associated with tumor progression and immune evasion. Our findings suggest a relationship between extracellular purines and the development and progression of the tumor microenvironment and poor outcomes. These findings increase the understanding of biological mechanisms related to HNC and demonstrate that these components are potential diagnostic markers and therapeutic targets for future management strategies and improvement in the quality of life.


Human food choices: an examination of the factors underlying acceptance/rejection of novel and familiar animal and nonanimal foods.
Two studies, aimed at increasing our knowledge of the factors that contribute to acceptance of novel foods were conducted. In Study 1, male and female participants were presented with familiar and novel animal and nonanimal foods and were asked to rate these foods on a variety of measures including: beliefs about the foods, feelings to the thought of ingesting the foods, and willingness to try the foods. Results demonstrated that willingness to try familiar nonanimal foods was predicted by beliefs about the negative sensory properties of the foods and joy elicited by the thought of eating them. Willingness to try familiar animal foods was predicted by beliefs about the sensory properties of the foods and beliefs about the anticipated negative consequences of eating them. Conversely, willingness to try novel foods, of both animal and nonanimal origin was predicted by beliefs about the disgusting properties of these foods and the interest evoked at the thought of consuming them. Study 2 used a qualitatively different sample from that used in Study 1 and perfectly replicated the previous results pertaining to the novel foods. Implications for our understanding of food-related disgust rejections are considered.


Intra- and extraneuronal formation of the two major noradrenaline metabolites in the cns of rats.
The two major noradrenaline metabolites in the rat brain, total 3-methoxy-4-hydroxyphenylglycol (MOPEG) and total 3,4-dihydroxyphenylglycol (DOPEG), were measured by a new analytical procedure after intraventricular (i.v.t.) injection of (3-H) noradrenaline or (3-H) dopamine. I.v.t. injection of (3-H) noradrenaline to rats with a 6-hydroxydopamine-induced destruction of nerve terminals in the central nervous system, resulted in an increased accumulation of 3-H-MOPEG compared to 3-H-DOPEG. On the contrary, reserpine induced a greater increase in 3-H-DOPEG than in 3-H-MOPEG accumulation when these two metabolites were formed from (3-H) noradrenaline synthesized in vivo by i.v.t. injection of (3-H) dopamine. These results indicate that the formation of DOPEG and MOPEG occur with some preference intra- and extraneuronally, respectively. The results, however, also show that these two formations sites should not be considered as specific.


Platelet-derived alpha-granules are associated with inflammation in patients with NK/T-cell lymphoma-associated hemophagocytic syndrome.
Due to the variable overlap of multiple symptoms, accurate early diagnosis of NK/T-cell lymphoma-associated hemophagocytic syndrome (NK/T-LAHS) is difficult, making the prognosis extremely poor. Hemophagocytic syndrome (HPS) is now diagnosed primarily based on the hemophagocytic lymphohistiocytosis (HLH)-2004 diagnostic criteria, and platelet count is one of the baseline evaluations. However, in our study, the data showed that decreased platelets were not only a clinical feature of HPS but also the key cells that regulate inflammation by releasing α-granules containing upregulated platelet factor 4 (PF4) and downregulated platelet-derived growth factors (PDGFs). Furthermore, we found that angiopoietin-4 (ANG-4), which has significant differential expression, has been less reported, that may affect hematopoiesis and proinflammatory responses and can be used as diagnostic biomarkers together with PF4 and PDGFs.


Regulation of macrophage phenotype by long-term exposure to IL-10.
Macrophages are chronically exposed to IL-10 in a variety of physiological and pathological settings. Macrophage responses to short-term stimulation with IL-10 have been extensively studied, but the effects of chronic exposure to IL-10 on macrophage function are not known. Herein we used transcriptional profiling and functional studies to characterize the phenotype of macrophages after long-term culture with IL-10. Classical activation of macrophages by LPS and IFNgamma was suppressed by IL-10. In contrast, IL-10 activated expression of genes that suggested acquisition of functions important for cell trafficking, tissue remodeling, recognition of microbial pathogens and responsiveness to the T cell-derived cytokines IL-2 and IL-21. These results demonstrate that IL-10 induces a differentiation program in macrophages and characterize a novel alternatively activated macrophage phenotype.


Literature mining of protein-residue associations with graph rules learned through distant supervision.
We propose a method for automatic extraction of protein-specific residue mentions from the biomedical literature. The method searches text for mentions of amino acids at specific sequence positions and attempts to correctly associate each mention with a protein also named in the text. The methods presented in this work will enable improved protein functional site extraction from articles, ultimately supporting protein function prediction. Our method made use of linguistic patterns for identifying the amino acid residue mentions in text. Further, we applied an automated graph-based method to learn syntactic patterns corresponding to protein-residue pairs mentioned in the text. We finally present an approach to automated construction of relevant training and test data using the distant supervision model. The performance of the method was assessed by extracting protein-residue relations from a new automatically generated test set of sentences containing high confidence examples found using distant supervision. It achieved a F-measure of 0.84 on automatically created silver corpus and 0.79 on a manually annotated gold data set for this task, outperforming previous methods. The primary contributions of this work are to (1) demonstrate the effectiveness of distant supervision for automatic creation of training data for protein-residue relation extraction, substantially reducing the effort and time involved in manual annotation of a data set and (2) show that the graph-based relation extraction approach we used generalizes well to the problem of protein-residue association extraction. This work paves the way towards effective extraction of protein functional residues from the literature.


Comparison of automated and manual methods for washing red blood cells.
We sought to compare the quality of washed red blood cells (RBCs) produced using the ACP215 device or manual methods with different combinations of wash and storage solutions. Our aim was to establish manual methods of washing that would permit a shelf life of more than 24 hours. Fourteen-day-old RBCs were pooled, split, and washed in one of five ways: 1) using the ACP215 and stored in SAGM, 2) manually washed and stored in saline, 3) manually washed in saline and stored in SAGM, 4) manually washed in saline-glucose and stored in SAGM, and 5) manually washed and stored in SAGM. Additional units were pooled and split, washed manually or using the ACP215, and irradiated on Day 14. Units were sampled to 14 days after washing and storage at 4 ± 2°C. All washed RBCs met specification for volume (200-320 mL) and hemoglobin (Hb) content (>40 g/unit). Removal of plasma proteins was better using manual methods: residual immunoglobulin A in saline-glucose-washed cells 0.033 (0.007-0.058) mg/dL manual versus 0.064 (0.026-0.104) mg/dL ACP215 (median, range). Hb loss was lower in manually washed units (mean, ≤ 2.0g/unit) than in ACP215-washed units (mean, 6.1 g/unit). Disregarding saline-washed and stored cells, hemolysis in all nonirradiated units was less than 0.8% 14 days after washing. As expected, the use of SAGM to store manually washed units improved adenosine triphosphate, glucose, lactate, and pH versus storage in saline. The data suggest that the shelf life of manually washed RBCs could be extended to 14 days if stored in SAGM instead of saline.


Awareness of the non-contraceptive benefits of reversible contraceptive methods in a cohort of Brazilian women: an exploratory study.
We assess the awareness of Brazilian women about non-contraceptive benefits of contraceptives and compare non-health with health care providers. We conducted a survey in Brazil using a questionnaire administered via Google Forms asking women to respond about the awareness of non-contraceptive benefits of combined oral contraceptives (COCs), depot-medroxyprogesterone acetate (DMPA), 52 mg levonorgestrel-releasing intrauterine system (LNG-IUS), etonogestrel (ENG)-implant, copper intrauterine device (Cu-IUD), and male condom. We received 2,068 completed questionnaires, 720 women (34.8%) aged ≤ 29 years and 236 (11.4%) were physicians or nurses. Only one third of the respondents were aware that COC use is associated with decreased risks of ovarian and endometrial cancers; 296 (16.1%) about that the use of DMPA is associated with a decreased risk of endometrial cancer, and 253 (13.8%) were aware about that the users of the Cu-IUD present lower risk of cervical cancer. We identified significant differences between non-health care providers when compared to physicians or nurses. We found low awareness regarding the associated lower risk of ovarian, endometrial and cervical cancer associated with the use of some contraceptives, evidencing the need to provide more information about the non-contraceptive benefits during training for health care providers.


Enhancer activity-based identification of functional enhancers using zebrafish embryos.
Chromatin immunoprecipitation (ChIP) against enhancer-associated marks with massive sequencing is a powerful approach to identify genome-wide distributions of putative enhancers. However, functional in vivo analysis is required to elucidate the activities of predicted enhancers. Using zebrafish embryos, we established a ChIP-Injection method that enables identification of functional enhancers based on their enhancer activities in embryos. Each reporter gene possessing the enhancer-associated genomic region enriched by ChIP was injected into zebrafish embryos to analyze the activity of putative enhancers. By using the ChIP-Injection, we identified 32 distinct putative enhancers that drove specific expression. Additionally, we generated transgenic lines that exhibit distributions of the EGFP signal as was observed in the screening. Furthermore, the expression pattern driven by the identified somite-specific enhancer resembled that of the gene acta2. The results indicate that ChIP-Injection provides an efficient approach for identification of active enhancers in a potentially wide variety of developmental tissues and stages.


The oligosaccharide chains of cobra venom factor are required for complement activation.
To examine the function of the carbohydrate chains of cobra venom factor (CVF), the molecule was enzymatically deglycosylated under non-denaturing conditions with N-glycanase (peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase). The deglycosylation of CVF chains seems to proceed independently of each other, leading to partially deglycosylated intermediates. Complete deglycosylation of CVF was found to abolish the activity of CVF. The deglycosylated molecule is unable to activate the alternative pathway of complement. Deglycosylated CVF no longer consumes the serum complement activity, it does not induce C3 activation in serum, nor does it induce complement-mediated hemolysis. These results indicate that the carbohydrate moieties of CVF are essential for its role in complement activation.


Prevalence and correlates of metabolic syndrome (MS) in older adults.
Metabolic syndrome (MS) is known to increase risks for cardiovascular disease. Risks for cardiovascular disease also increase with aging. The aim of the study was to describe prevalence and correlates of MS in older adults. Patients aged 65 years and over without any acute illness that were referred to our geriatrics unite for comprehensive geriatric assessment were included in this cross-sectional study. MS was defined by using the WHO and the National Cholesterol Education Program (NCEP) definitions. The correlates were age, gender, low-grade inflammation as assessed by C-reactive protein (CRP) levels, high-homocysteine, total and LDL cholesterol, lipoprotein-a (Lip-a), apolipoprotein-A (Apo-A), apolipoprotein-B (Apo-B), nutrition point, coronary artery disease (CAD) and cerebrovascular event history. Total 1255 patients, 789 (62.9%) females, 466 (37.1%) males with a mean age of 71.8+/-6.3 years were included in our study. MS prevalence was 16.2% with WHO definition and 23.8% with NCEP definition. Prevalence of CAD in MS and non-metabolic syndrome (non-MS) patients was 38.4% versus 29.5% (p=0.010) in WHO and 35.3% versus 29.6% (p=0.066) in NCEP group, respectively. Prevalence of cerebrovascular event history was 11.3% versus 6.2% in WHO (p=0.008) and 9.9% versus 6.1% (p=0.026) in NCEP group, respectively. Multiple logistic regression analysis was performed to investigate the independent association of variables with the MS. Female sex, high-homocysteine, low-grade inflammation, CAD and cerebrovascular event history was found to be associated with both modified WHO and NCEP MS groups in the multivariate analysis. This study has shown that MS is common in elderly patients with strongly related to CAD and cerebrovascular events. Low grade inflammation as assessed by CRP and high-homocysteine level is strongly related to MS.


The slope method: a tool for analyzing semi-continuous data.
The MixAlco process is a biorefinery that converts lignocellulose into useful chemicals and hydrocarbon fuels via mixed-acid fermentation. For a semi-continuous-staged fermentation train, during each transfer, discrete amounts of material are moved between fermentors and data are tabulated. Because of natural day-to-day variations, the data are inherently noisy. To calculate performance parameters (e.g., yield, conversion, selectivity, productivity), the average flowrate of each stream component must be determined. To minimize error associated with noise, three data analysis methods were compared: Average, Accumulation, and Slope. The Average method determines the flowrate by averaging the amounts moved each transfer. The Accumulation method stores the solids and liquids that exit the fermentation train in separate vessels. After an extended time period, the mass in each storage vessel is measured so the average flowrate can be calculated. The Slope method calculates the flowrate of material in each stream from the slope of the moving cumulative sum with respect to time. For all three methods, the measured rates were virtually identical; thus accuracy was not affected by the method. However, for the examples presented, the Average method had >40% error and the Slope method <4% error; thus, precision was significantly affected by the method. The Accumulation method calculated the flowrate with a single data point so it is not possible to determine the statistical error.


The role of dopamine in drug abuse viewed from the perspective of its role in motivation.
Drugs of abuse share with conventional reinforcers the activation of specific neural pathways in the CNS that are the substrate of their motivational properties. Dopamine is recognized as the transmitter of one such neural pathway, being involved in at least three major aspects of motivation: modulation of motivational state, acquisition (incentive learning) and expression of incentive properties by motivational stimuli. Drugs of abuse of different pharmacological classes stimulate in the low dose range dopamine transmission particularly in the ventral striatum. Apart from psychostimulants, the evidence that stimulation of dopamine transmission by drugs of abuse provides the primary motivational stimulus for drug self-administration is either unconvincing or negative. However, stimulation of dopamine transmission is essential for the activational properties of drugs of abuse and might be instrumental for the acquisition of responding to drug-related incentive stimuli (incentive learning). Dopamine is involved in the induction and in the expression of behavioural sensitization by repeated exposure to various drugs of abuse. Sensitization to the dopamine-stimulant properties of specific drug classes leading to facilitation of incentive learning of drug-related stimuli might account for the strong control over behaviour exerted by these stimuli in the addiction state. Withdrawal from drugs of abuse results in a reduction in basal dopamine transmission in vivo and in reduced responding for conventional reinforcers. Although these changes are likely to be the expression of a state of dependence of the dopamine system their contribution to the motivational state of drug addiction is unclear.


Ultrastructural localization of a lysosomal enzyme in resin-embedded lymphocytes.
The intracellular distribution of lysosomal enzymes in lymphocytes has previously been only poorly defined, mainly by cytochemical procedures of low resolution. In the present study we have used a post-embedding immunogold technique to identify the precise ultrastructural localization of a lysosomal enzyme, beta-glucuronidase, in activated lymphocytes embedded in Lowicryl K4M resin. We show that this enzyme is present in the rough endoplasmic reticulum, in the Golgi complex, and in vesicular organelles which probably include lysosomes.


Proteomic analysis of the left ventricle post-myocardial infarction to identify in vivo candidate matrix metalloproteinase substrates.
Left ventricular remodeling post-myocardial infarction (MI) involves a multitude of mechanisms that regulate the repair response. Matrix metalloproteinases (MMPs) are a major family of proteolytic enzymes that coordinate extracellular matrix turnover. MMP-7 or MMP-9 deletion attenuate adverse remodeling post-MI, but the mechanisms have not been fully clarified. Both MMP-7 and MMP-9 have a large number of known in vitro substrates, but in vivo substrates for these two MMPs in the myocardial infarction setting are incompletely identified. Advances in proteomic techniques have enabled comprehensive profiling of protein expression in cells and tissue. In this chapter, we describe a protocol for the proteomic analysis of in vivo candidate MMP substrates in the post-MI left ventricle using two-dimensional electrophoresis, liquid chromatography coupled with tandem mass spectrometry, and immunoblotting.


Aromatase inhibitors and breast cancer.
Selective aromatase inhibitors cause regression of breast carcinomas by reducing estrogen production via inhibition of the enzyme aromatase (estrogen synthetase). A higher incidence of hormone-dependent breast cancer occurs in postmenopausal women than in younger women. Thus, total estrogen blockade is more likely to be achieved with systemic (pharmacologic) methods than by surgical removal of endocrine glands. At the present time, breast cancer patients with hormone-dependent disease usually receive the antiestrogen tamoxifen as first-line treatment. Although tamoxifen is effective initially, patients develop resistance to the drug, which results in disease progression. Aromatase inhibitors acting by a different mechanism from tamoxifen are effective in some of these patients. There is also the potential that aromatase inhibitors could be more effective as first-line treatment. Formestane, 4-hydroxyandrostenedione, the first approved aromatase inhibitor, is proving to be useful in postmenopausal breast cancer patients with advanced disease. Recently, anastrozole, a nonsteroidal agent, has been approved in the United States. Other very potent aromatase inhibitors are under development. Aromatase inhibitors as a new class of well-tolerated agents are now becoming available for improving the treatment of breast cancer patients.


Nano-crystalline cellulose: Preparation, modification and usage as sustained release drug delivery excipient for some non-steroidal anti-inflammatory drugs.
Nano crystalline cellulose (NCC) has gained attention in the pharmaceutical industry owing to its biodegradable nature and high mechanical properties. In the present work we reported preparation of NCC from cellulose isolated from a bio-waste, its modification and exploration as drug delivery excipient for sustained release of non-steroidal anti-inflammatory drugs (NSAIDs). Cellulose extracted from Citrus limetta albedo was subjected to alkali treatment, bleaching and hydrolysis by sulfuric acid to obtain NCC of particle size ranging from 1 to 10 nm. Further, modification with a cationic surfactant cetyltrimethylammonium bromide (CTAB) resulted in NCC-CTAB with enhanced surface area and high aspect ratio which was investigated as sustained release drug delivery system (DDS) for NSAID. Drug loading capacity correlated well with log P value of NSAID. NSAID-loaded NCC-CTAB acted as sustained release DDS over a period of 3hr. The results were confirmed as serum protein protecting and anti-cathepsins activities. CTAB-NSAID interactions have been assessed using CHEM3D.


Combined hepatic resection with the inferior vena cava and diaphragm and reconstruction using an equine pericardial patch: report of a case.
We herein report a case of combined hepatic resection with inferior vena cava (IVC) and diaphragm resection, and reconstruction using an equine pericardial patch. A 54-year-old woman showed hepatic cancer recurrence on radiological imaging, with invasion to the caudate lobe of the liver, IVC, diaphragm, and adrenal gland. We resected 10 × 5 cm of the diaphragm. After dissecting the hepatic parenchyma, the caudate lobe was connected only to the IVC. Clamping of the IVC was performed between the IVC below the confluence of the hepatic vein and the suprarenal IVC. A 6 × 3-cm segment of the IVC was then resected. The IVC and diaphragm were reconstructed using an equine pericardial patch, as both defects were too large to repair without a patch. The equine pericardium represents a suitable graft material for repairing both the IVC and diaphragm. Further investigation is needed to determine the durability and anti-infection properties of equine pericardial grafts.


Environmental enrichment promotes robust functional and histological benefits in female rats after controlled cortical impact injury.
Environmental enrichment (EE) consistently induces marked benefits in male rats after traumatic brain injury (TBI), but whether similar efficacy extends to females is not well established. Hence, the aim of this study was to reassess the effect of EE on functional and histological outcome in female rats after brain trauma. Twenty-four normal cycling adult female rats underwent verification of estrous stage prior to controlled cortical impact (CCI) or sham injury and then were assigned to EE or standard (STD) housing. Motor function was assessed with beam-balance/beam-walk and rotarod tasks on post-operative days 1-5 and every other day from 1-19, respectively. Spatial learning/memory was evaluated in a Morris water maze on days 14-19. Morphologically intact hippocampal CA(1/3) cells and cortical lesion volume were quantified 3 weeks after injury. No differences were observed between the EE and STD sham groups in any endpoint measure and thus the data were pooled. In the TBI groups, EE improved beam-balance, beam-walk, rotarod, and spatial learning performance vs. STD (p's<0.05). EE also provided significant histological protection as confirmed by increased CA(1/3) cell survival and decreased cortical lesion size vs. STD. These data demonstrate that EE confers robust benefits in female rats after CCI injury, which parallels numerous studies in males and lends further credence for EE as a preclinical model of neurorehabilitation.


Influence of tumor cell-derived TGF-β on macrophage phenotype and macrophage-mediated tumor cell invasion.
In oral squamous cell carcinoma (OSCC), macrophages are the most abundant immune cell type in the tumor microenvironment (TME). Macrophage infiltration is inversely proportional to prognosis and disease survival, particularly when these tumor-associated macrophages (TAM) assume an M2-like phenotype. This phenotype is determined by cues from the microenvironment, especially tumor cell-secreted molecules, and is associated with increased production of extracellular-matrix-degrading enzymes, angiogenic molecules and immunosuppressing cytokines. This study investigates, in vitro and in vivo, the relative contribution of OSCC cell-secreted transforming growth factor beta (TGF-β) on the phenotype of macrophages and on macrophage-facilitated tumor invasion. TCGA database shows a positive correlation between high expression of TGFB1 and macrophage infiltrate in Head and neck squamous cell carcinoma (HNSCC). THP-1 derived-macrophages were exposed to the secretome of two OSCC cell lines using two strategies to block the effects of neoplastic cell-secreted TGF-β: pre-treatment with a TGF-β receptor type I kinase inhibitor (LY364947) and antibody-mediated depletion. RT-qPCR, ELISA and flow cytometry determined macrophage phenotype after exposure to conditioned medium (CM) from H-314 (TGF-βhigh) or SCC-9 (TGF-βlow) cell lines. The influence of TGF-β on macrophage-mediated tumor cell invasion (myogel and CAM assays) and chemotaxis (Boyden chamber) was assessed using co-cultures of macrophages and OSCC cells in which macrophages were pre-conditioned with the secretome of OSCC cells in the presence and absence of LY364947. Blocking the effects of TGF-β skewed macrophages to the M1 end of the phenotype by differential effects depending on the strategy for inhibiting the influence of TGF-β and on the neoplastic cell secretome. In vitro and in vivo invasion of H-314 cell line was reduced by inhibiting TGFBR1 signaling in macrophages, whereas SCC-9 cell invasion was not affected. SCC-9/macrophage reciprocal chemotaxis were enhanced by inhibiting TGFBR1 signaling in macrophages, whereas only macrophage chemotaxis to H314 products was inhibited by inhibiting TGFBR1. In summary, blocking the effects of OSCC cell-secreted TGF-β in macrophages attenuates M2-like phenotypical traits of macrophages and can impact invasion and chemotaxis of tumor cells differentially.


LAP proteins: what's up with epithelia?
How cells maintain their overall shape and size, and the related question of how proteins and other molecules reach and stay at their specific subcellular locations, are among the most difficult and exciting problems in cell biology. Three recent studies have made a significant contribution to this area by identifying new proteins, called LAP proteins, that have critical functions in maintaining the shape and apical-basal polarity of epithelial cells.