Calculation of electron dose to target cells in a complex environment by Monte Carlo code "CELLDOSE".
We used the Monte Carlo code "CELLDOSE" to assess the dose received by specific target cells from electron emissions in a complex environment. (131)I in a simulated thyroid was used as a model. Thyroid follicles were represented by 170 microm diameter spherical units made of a lumen of 150 microm diameter containing colloidal matter and a peripheral layer of 10 microm thick thyroid cells. Neighbouring follicles are 4 microm apart. (131)I was assumed to be homogeneously distributed in the lumen and absent in cells. We firstly assessed electron dose distribution in a single follicle. Then, we expanded the simulation by progressively adding neighbouring layers of follicles, so to reassess the electron dose to this single follicle implemented with the contribution of the added layers. Electron dose gradient around a point source showed that the (131)I electron dose is close to zero after 2,100 microm. Therefore, we studied all contributions to the central follicle deriving from follicles within 12 orders of neighbourhood (15,624 follicles surrounding the central follicle). The dose to colloid of the single follicle was twice as high as the dose to thyroid cells. Even when all neighbours were taken into account, the dose in the central follicle remained heterogeneous. For a 1-Gy average dose to tissue, the dose to colloidal matter was 1.168 Gy, the dose to thyroid cells was 0.982 Gy, and the dose to the inter-follicular tissue was 0.895 Gy. Analysis of the different contributions to thyroid cell dose showed that 17.3% of the dose derived from the colloidal matter of their own follicle, while the remaining 82.7% was delivered by the surrounding follicles. On the basis of these data, it is shown that when different follicles contain different concentrations of (131)I, the impact in terms of cell dose heterogeneity can be important. By means of (131)I in the thyroid as a theoretical model, we showed how a Monte Carlo code can be used to map electron dose deposit and build up the dose to target cells in a complex multi-source environment. This approach can be of considerable interest for comparing different radiopharmaceuticals as therapy agents in oncology.


Effect of glycemic index meals on recovery and subsequent endurance capacity.
This study examined the effect of ingesting a high or low glycemic index (GI) meal during a short-term recovery period on endurance running capacity. On two occasions, seven men (age 30.0+/-2.6 yr, body mass 60.7+/-1.4 kg, VO (2max) 62.1+/-2.2 ml.kg (-1).min (-1)) ran at 70% VO (2max) on a level treadmill for 90 min (R1), followed by a 4 h recovery (REC) and a further exhaustive run at the same speed (R2). Twenty minutes after R1, each subject consumed an isoenergetic meal containing either high GI (HGI, GI=77) or low GI (LGI, GI=37) carbohydrate providing 1.5 g CHO.kg (-1) BM. During REC, subjects also ingested a prescribed volume of water equal to 150% of their BM loss during R1. The duration of R2 in the HGI trial was 15% longer than in the LGI trial (HGI: 86.6+/-10.7 min vs. LGI: 75.2+/-8.1 min, p<0.05). The subjects also achieved complete rehydration after REC on both occasions. In conclusion, the consumption of a HGI meal during a 4 h recovery improved endurance capacity in a subsequent run; however, the precise mechanism(s) by which this takes place is yet to be clarified.


Clinical significance of CD44 variant 9 expression as a prognostic indicator in bladder cancer.
CD44, a major surface receptor for hyaluronic acid, has multiple isoforms and represents a major cancer stem cell marker for various epithelial tumors. CD44 variant 9 (CD44v9) was correlated with recurrence and metastasis in gastric and colon cancer. We examined its role in invasion and as a biomarker for the basal muscle invasive molecular subtype showing worse prognosis, and for tumor progression in high risk (pT1/high grade) non‑muscle invasive bladder cancers (NMIBCs). CD44v9, cytokeratin 5/6 (CK5/6), and cytokeratin 20 (CK20) expression was evaluated by immunohistochemistry in 98 pathologically confirmed specimens (36 muscle and 62 high‑risk non‑muscle) and correlated to clinical outcome. In vitro analysis was performed using two human bladder cancer cell lines (HT1376 and 5637). The CD44v9 high‑expressing group exhibited significantly lower progression‑free and cancer‑specific survival rates in both muscle (P=0.0349 and 0.0382, respectively) and non‑muscle (P=0.0002 and 0.0079) invasive patients. CD44v9 expression was significantly correlated with CK5/6 (P<0.001), colocalizing at the muscle invasion front but distinctly separated from CK20 in non‑muscle invasion. CD44 and CD44v9 siRNA knockdown demonstrated significantly lower Matrigel invasion ability and significantly shorter migration distance (all P<0.01). CD44 and CD44v9 knockdown increased E‑cadherin and decreased N‑cadherin, snail, and slug epithelial‑mesenchymal transition marker protein expression. Thus, higher CD44v9 expression was associated with worse prognosis, likely impacting invasion and migration via the epithelial‑mesenchymal transition. Together, these findings suggest that CD44v9 expression might be a useful predictive biomarker in basal‑type muscle and high-risk NMIBC.


A refined model of the prototypical Salmonella SPI-1 T3SS basal body reveals the molecular basis for its assembly.
The T3SS injectisome is a syringe-shaped macromolecular assembly found in pathogenic Gram-negative bacteria that allows for the direct delivery of virulence effectors into host cells. It is composed of a "basal body", a lock-nut structure spanning both bacterial membranes, and a "needle" that protrudes away from the bacterial surface. A hollow channel spans throughout the apparatus, permitting the translocation of effector proteins from the bacterial cytosol to the host plasma membrane. The basal body is composed largely of three membrane-embedded proteins that form oligomerized concentric rings. Here, we report the crystal structures of three domains of the prototypical Salmonella SPI-1 basal body, and use a new approach incorporating symmetric flexible backbone docking and EM data to produce a model for their oligomeric assembly. The obtained models, validated by biochemical and in vivo assays, reveal the molecular details of the interactions driving basal body assembly, and notably demonstrate a conserved oligomerization mechanism.


Uninformative memories will prevail: the storage of correlated representations and its consequences.
Autoassociative networks were proposed in the 80's as simplified models of memory function in the brain, using recurrent connectivity with Hebbian plasticity to store patterns of neural activity that can be later recalled. This type of computation has been suggested to take place in the CA3 region of the hippocampus and at several levels in the cortex. One of the weaknesses of these models is their apparent inability to store correlated patterns of activity. We show, however, that a small and biologically plausible modification in the "learning rule" (associating to each neuron a plasticity threshold that reflects its popularity) enables the network to handle correlations. We study the stability properties of the resulting memories (in terms of their resistance to the damage of neurons or synapses), finding a novel property of autoassociative networks: not all memories are equally robust, and the most informative are also the most sensitive to damage. We relate these results to category-specific effects in semantic memory patients, where concepts related to "non-living things" are usually more resistant to brain damage than those related to "living things," a phenomenon suspected to be rooted in the correlation between representations of concepts in the cortex.


Acute radiologic intervention in gastrointestinal bleeding.
A case of embolization of the gastroduodenal artery in a 38-year old man with chronic pancreatitis and uncontrollable bleeding is presented. The advantage of this interventional radiologic procedure is discussed and in selective cases it seems to be the choice of treatment.


Foregut duplication cysts in the head and neck: presentation, diagnosis, and management.
To review the presentation, diagnosis, and management of foregut duplication cysts of the head and neck in our institution. An institutional review board-approved retrospective review of all patients treated for foregut duplication cysts of the head and neck over an 18-year period. Pediatric otolaryngology tertiary referral center. Twenty-two patients with 23 pathologically confirmed foregut duplication cysts of the head and neck were identified. Fourteen patients (64%) were male. The median age at diagnosis was 1.5 years (age range, 5 days to 7 years). Clinical data, including age, presenting symptoms, anatomical site(s), evaluation, treatment, and complication, were recorded and analyzed. Presentation varied depending on anatomical site of involvement, with 12 patients (55%) being asymptomatic. The cysts were found in the oral cavity (n = 12), oropharynx (n = 6), supraglottis (n = 2), and neck (n = 3). Imaging, which was performed in 13 patients and consisted of magnetic resonance imaging (n = 8), computed tomography (n = 5), and ultrasonography (n = 1), demonstrated the cystic nature of the lesions. All patients underwent surgical excision, which focused on excising the cyst, while preserving surrounding normal tissues. No patient demonstrated recurrence at follow-up. Foregut duplication cysts of the head and neck, although uncommon, should be included in the differential diagnosis of cystic head and neck lesions. Preoperative imaging is recommended to differentiate these lesions from other congenital head and neck masses. Surgical excision biopsy with complete removal of the mucosal lining is curative, with no instances of recurrence in our series.


[Effects of electroacupuncture on IL-2-IFN-NKC immunity immunoloregulation net and IL-2 receptor in rats with exercise stress].
To explore the effects of electroacupuncture on exercise-induced immunosuppression in rats and its mechanism. Sports immunosuppressive model was established successfully by the rats were conducted high intensity swimming training 150 min/day, 6 days/wk for 8 weeks in this study. Forty-three SD rats were randomly divided into a control group (group A, n = 10), a high intensity swimming training group (group B, n = 17), and a high intensity plus electroacupuncture group (group C, n = 16). Group A did not receive any intervention. Group B was conducted 150 min/day, 6 days/wk swimming training for 8 weeks. Group C was treated with electroacupuncture at "Baihui" (GV 20), "Guanyuan" (CV 4) and "Zusanli" (ST 36) after every exercise-time from the second week, once each day for 7 weeks. The changes of the rats' weight, gamma-interferon (gamma-IFN), interleukin-2 (IL-2), solubility IL-2 receptor (SIL-2R) and nature killer cell (NKC) were detected. (1) Compared with group A, gamma-IFN and IL-2 in group B were significantly decreased (P < 0.01, P < 0.05), and NKC in group C was significantly increased (P < 0.01). Meanwhile, gamma-IFN and NKC in group C were both significantly higher than that in group B (P < 0.05, P < 0.01). (2) Compared with group A, the weight of the rats in group B and group C were significantly decreased (both P < 0.01), but SIL-2R in group B was significantly increased (P < 0.05). The weight of the rats in group C was significantly higher than that in group B (P < 0.05) and SIL-2R in group C was significantly lower than that in group B (P < 0.01). Lasting gravis exercise stress does decrease the immune function in rats and is even inhibited significantly, but electroacupuncture can up-regulate the exercise-induced immunosuppression.


Spindle and Kinetochore-associated Family Genes are Prognostic and Predictive Biomarkers in Hepatocellular Carcinoma.
Hepatocellular carcinoma (HCC) is one of the most frequent malignant tumors. Spindle and kinetochore-associated (SKA) family genes are essential for the maintenance of the metaphase plate and spindle checkpoint silencing during mitosis. Recent studies have indicated that dysregulation of SKA family genes induces tumorigenesis, tumor progression, and chemoresistance via modulation of cell cycle and DNA replication. However, the differential transcription of SKAs in the context of HCC and its prognostic significance has not been demonstrated. Bioinformatics analyses were performed using TCGA, ONCOMINE, HCCDB, Kaplan-Meier plotter, STRING, GEPIA databases. qRT-PCR, western blot, and functional assays were utilized for in vitro experiments. We found remarkable upregulation of transcripts of SKA family genes in HCC samples compared with normal liver samples on bioinformatics analyses and in vitro validation. Interaction analysis and enrichment analysis showed that SKA family members were mainly related to microtubule motor activity, mitosis, and cell cycle. Immuno-infiltration analysis showed a correlation of all SKA family genes with various immune cell subsets, especially T helper 2 (Th2) cells. Transcriptional levels of SKA family members were positively associated with histologic grade, T stage, and α-fetoprotein in HCC patients. Receiver operating characteristic curve analysis demonstrated a strong predictive ability of SKA1/2/3 for HCC. Increased expression of these SKAs was associated with unfavorable overall survival, progression-free survival, and disease-specific survival. On Cox proportional hazards regression analyses, SKA1 upregulation and pathological staging were independent predictors of overall survival and disease-specific survival of HCC patients. Finally, clinical tissue microarray validation and in vitro functional assays revealed SKA1 acts an important regulatory role in tumor malignant behavior. SKA family members may potentially serve as diagnostic and prognostic markers in the context of HCC. The correlation between SKAs and immune cell infiltration provides a promising research direction for SKA-targeted immunotherapeutics for HCC.


Low blood pressure and preserved systolic function in elders with heart failure.
This literature review was performed to evaluate the relationship of systolic blood pressure (SBP) and preserved systolic function on morbidity and mortality in individuals older than 65 years with heart failure (HF). When prolonged, high SBP, defined as measurements greater than 140 to 160 mm Hg, is associated with increased risk of developing HF. Medications to lower SBP measurements to Joint National Committee VII goals of less than 140 mm Hg are often prescribed on the assumption that treatment guidelines result from a systematic analysis of clinical trials and efficacy of drug treatments. Lower limits of SBP are less defined in current guidelines optimizing HF. The electronic databases PubMed, CINAHL, and Google Scholar are searched for keywords heart failure, prognosis, preserved systolic function, and blood pressure (BP). Five scholarly research articles investigated the effects of variables, notably SBP and confirmed preserved systolic function, on HF, published in the English language between July 2006 and January 2009. Inclusion criteria were study samples consisting of individuals 65 years and older referred in this article as elders, with the diagnosis of HF with a focus on variables measuring SBP and systolic function when assessing outcomes. Low SBP is a risk factor for adverse outcomes in HF elders. Additionally, female elders more commonly had preserved systolic function, but presented with less classic symptoms of HF and were less likely to receive cardiology consultation. Considerations for future research are the inclusion of participants presenting with SBP of less than 110 mm Hg in clinical trials and updated evidence-based guidelines, defining acceptable increased target BP ranges, for sex- and age-adjusted HF patients with preserved systolic function.


Assessing health-related quality of life pre- and post-liver transplantation: a prospective multicenter study.
We report on a prospective multicenter study to assess pretransplantation and posttransplantation health-related quality of life (HRQL) of liver transplant recipients. HRQL was assessed at several timepoints using a self-completion questionnaire consisting of the EuroQol instrument (EQ-5D) and the 36-Item Short-Form Health Survey (SF-36) health status profile measure. All individuals selected to receive treatment as part of the UK NHS liver transplantation program at each of six liver transplantation centers in England and Wales during a 2-year period (n = 542) were eligible to be included on the study. An overall response rate of 84% (455 responses) was achieved. A paired comparison of HRQL at listing and 3 months posttransplantation showed statistically significant improvements (P <.05) in all dimensions of the SF-36 (with the exception of Bodily Pain [P =.686]) and the EQ-5D tariff and visual analogue scale (VAS) scores. An analysis of posttransplantation HRQL over time for patients who survived until the end of the study (24 months posttransplantation) showed a statistically significant improvement (P <.05) for all dimensions of the SF-36 (apart from Mental Health [P =.245] and Role-Emotional dimensions [P =.265]) and the EQ-5D VAS and tariff scores. Adjusting for patients who died posttransplantation reduced mean EQ-5D tariff scores substantially, and the change over time in EQ-5D tariff scores was no longer statistically significant (P =.55). Results of regression analysis conducted to assess the importance of patient characteristics and center size on EQ-5D tariff and VAS scores generated posttransplantation indicated there was variation in scores according to patient age and center size.


Gastrointestinal processing of Na+, Cl-, and K+ during digestion: implications for homeostatic balance in freshwater rainbow trout.
The role of the gastrointestinal tract in maintaining ionic homeostasis during digestion, as well as the relative contribution of the diet for providing electrolytes, has been generally overlooked in many aquatic species. An experimental diet that contained an inert reference marker (lead-glass beads) was used to quantify the net transport of Na(+), K(+), and Cl(-) during the digestion and absorption of a single meal (3% ration) by freshwater rainbow trout (Oncorhynchus mykiss). Secretion of Cl(-) into the stomach peaked at 8 and 12 h following feeding at a rate of 1.1 mmol.kg(-1).h(-1), corresponding to a theoretical pH of 0.6 in the secreted fluid (i.e., 240 mmol/l HCl). The majority ( approximately 90%) of dietary Na(+) and K(+) was absorbed in the stomach, whereas subsequent large fluxes of Na(+) and Cl(-) into the anterior intestine corresponded to a large flux of water previously observed. The estimated concentration of Na(+) in fluids secreted into the anterior intestine was approximately 155 mmol/l, equivalent to reported hepatic bile values, whereas the estimated concentration of Cl(-) ( approximately 285 mmol/l) suggested seepage of HCl acid from the stomach in advance of the chyme front. Net absorption of K(+) in the stomach occurred following the cessation of Cl(-) secretion, providing indirect evidence of K(+) involvement with HCl acid production. Overall, 80-90% of the K(+) and Cl(-) contents of the meal were absorbed on a net basis, whereas net Na(+) absorption was negligible. Chyme-to-plasma ion concentration gradients were often opposed to the direction of ion transport, especially for Na(+) and Cl(-).


Latest Advances in Protein-Recovery Technologies from Agricultural Waste.
In recent years, downstream bioprocessing industries are venturing into less tedious, simple, and high-efficiency separation by implementing advanced purification and extraction methods. This review discusses the separation of proteins, with the main focus on amylase as an enzyme from agricultural waste using conventional and advanced techniques of extraction and purification via a liquid biphasic system (LBS). In comparison to other methods, such as membrane extraction, precipitation, ultrasonication, and chromatography, the LBS stands out as an efficient, cost-effective, and adaptable developing method for protein recovery. The two-phase separation method can be water-soluble polymers, or polymer and salt, or alcohol and salt, which is a simpler and lower-cost method that can be used at a larger purification scale. The comparison of different approaches in LBS for amylase purification from agricultural waste is also included. Current technology has evolved from a simple LBS into microwave-assisted LBS, liquid biphasic flotation (LBF), thermoseparation (TMP), three-phase partitioning (TPP), ultrasound-assisted LBS, and electrically assisted LBS. pH, time, temperature, and concentration are some of the significant research parameters considered in the review of advanced techniques.


Determinants of Drug Resistance in B-Cell Non-Hodgkin Lymphomas: The Case of Lymphoplasmacytic Lymphoma/Waldenström Macroglobulinemia.
Lymphoplasmacytic lymphoma (LPL) is a rare subtype of B cell-derived non-Hodgkin lymphoma characterized by the abnormal growth of transformed clonal lymphoplasmacytes and plasma cells. This tumor almost always displays the capability of secreting large amounts of monoclonal immunoglobulins (Ig) of the M class (Waldenström Macroglobulinemia, WM). The clinical manifestations of WM/LPL may range from an asymptomatic condition to a lymphoma-type disease or may be dominated by IgM paraprotein-related symptoms. Despite the substantial progresses achieved over the last years in the therapy of LPL/WM, this lymphoma is still almost invariably incurable and exhibits a propensity towards development of refractoriness to therapy. Patients who have progressive disease are often of difficult clinical management and novel effective treatments are eagerly awaited. In this review, we will describe the essential clinical and pathobiological features of LPL/WM. We will also analyze some key aspects about the current knowledge on the mechanisms of drug resistance in this disease, by concisely focusing on conventional drugs, monoclonal antibodies and novel agents, chiefly Bruton's Tyrosine Kinase (BTK) inhibitors. The implications of molecular lesions as predictors of response or as a warning for the development of therapy resistance will be highlighted.


Temozolomide treatment in glioma--experiences in two university hospitals in Finland.
Thirty-two patients with relapsing glioma were treated with temozolomide in two university hospitals in Finland. One patient (3%) had complete response and 9 (28%) partial response, with 8 patients (25%) showing stable disease. Median progression-free survival for these 18 patients (56%) was 7 months (range 2-11+). The remaining either had progressive disease (25%) or only clinical evaluation (19%). Karnofsky score improved in 34% of patients and decreased in 3%. Symptoms were alleviated in 44% and deteriorated in 9%. Grade 3-4 toxicity was detected in 9% of the patients. Only 4% of the days in treatment were spent in hospital. An average 1.8 neuroradiological investigations, 6.9 laboratory visits, and 5.3 visits to the oncologist were made. This study confirms that temozolomide has positive effects on the outcome of often heavily pretreated glioma patients. High drug costs are compensated by prolonged home care and even the possibility to maintain working capacity.


Ureteroscopy and stone lithotripsy with lithoclast: personal experience.
Ureteroscopy has become a common technique in the diagnosis and treatment of ureteral pathologies, but this procedure is quite invasive and some complications have been reported in literature. In our Institute 49 patients underwent ureteroscopy and ballistic lithotripsy with lithoclast for ureteral stones. The stones were localized both in the middle and distal part of the ureter. We used a small caliber 7-8.5 Wolf ureteroscope. The treatments were performed under antibiotic prophylaxis. Direct access to the ureter without dilation of the meatus was obtained in 97.96% of patients. The stones were easily reached in 93.88% of the cases and satisfactory fragmentation was obtained in 90.7%. In 4 patients (9.3%) one or more large stone fragments escaped into the kidney, requiring the patients to be treated with ESWL. No major complications occurred: no ureteral perforations, no important bleeding and no severe or persistent infections. All patients were discharged in one to four days postoperatively. The authors conclude that ureteroscopy using small caliber instruments with Lithoclast is a safe and satisfactory alternative to ESWL in the treatment of ureteral stones.


Cardiovascular effects of cocaine.
This article reviews the chemical nature of cocaine, an increasingly abused stimulant. Following a description of the clinical manifestations of drug use, cardiovascular effects as well as routes of administration are described. Clinical complications of acute and chronic cocaine abuse include dysrhythmias, acute myocardial ischemia and infarction, sudden death, myocarditis, and cardiomyopathy. Clinical manifestations and assessment protocols are summarized. Treatment protocols remain focused on symptom presentation at this time, as there is no known antidote for cocaine toxicity or overdose.


Effect of group thawing on post-thaw viability of bovine spermatozoa packaged in .5-milliliter French straws.
The objective of this study was to determine the effects of thawing groups of 2, 5, 10, 15, or 20 .5-ml French straws on post-thaw spermatozoal viability. Thermostatically controlled and nonthermostatically controlled thawing baths were compared. Using a split-plot design, semen from 10 bulls was extended in egg yolk citrate, frozen, and then thawed (in the respective groups) at 36 degrees C in two types of thawing baths. Motility and percentage of intact acrosomes were determined immediately after thawing (0 h) and again after 4 h of incubation at the respective temperature of each thawing bath. Neither percentage of intact acrosomes nor motility was influenced by the number of straws thawed at 0 h (P greater than .05). Thawing bath had no effect (P greater than .05) on motility or percentage of intact acrosomes at 0 h. Bull variation was significant in both the 0- and 4-h evaluations. After 4 h of incubation, there was a significant (P less than .05) straw number x thawing bath interaction. When 15 or 20 straws were thawed in the thermostatically controlled bath there was a reduction (P less than .05) in motility and percentage of intact acrosomes. However, in the nonthermostatically controlled bath there was no reduction in motility and percentage of intact acrosomes as the size of straw group increased. Our results indicate that, when using a nonthermostatically controlled thawing bath, semen packaged in .5-ml straws can be thawed in groups of 20 without an effect on post-thaw sperm viability.


CCAAT/enhancer-binding protein decoy oligodeoxynucleotide inhibition of macrophage-rich vascular lesion formation in hypercholesterolemic rabbits.
Many cytokine genes, including those encoding acute-phase proteins and immunoglobulins, share binding sites for the CCAAT/enhancer-binding protein (C/EBP) in their 5'-flanking regions, and C/EBP-related transcription factors regulate cell proliferation during terminal differentiation. Therefore, C/EBP represents an attractive target for inhibiting restenosis after balloon angioplasty. In a rabbit model of restenosis that combines balloon injury of the carotid artery with cholesterol-mediated chronic inflammation, a decoy oligodeoxynucleotide (ODN) capable of neutralizing C/EBP was administered to the site of injury for 30 minutes. Electrophoretic mobility shift analysis confirmed that C/EBP activity in decoy ODN-treated segments was virtually absent after 2 days. Morphometric analysis after 28 days revealed significant reduction (up to 50%) of neointimal formation and intravascular inflammation in decoy ODN-treated segments compared with mutant control ODN or vehicle-treated segments. In addition, de novo synthesis of endothelin-1 and the number of proliferating cell nuclear antigen-positive smooth muscle cells in the vessel wall were markedly attenuated at day 3. These findings suggest that decoy ODN-based neutralization of C/EBP may be a feasible and effective method to limit restenosis after angioplasty brought about, at least in part, by inhibiting the de novo synthesis of endothelin-1.


[Investigating the inhibitory effects of interferon-alpha on the replication of hepatitis C virus replicon].
To investigate the inhibitory effects of interferon-alpha (IFN-alpha) on HCV replicon and evaluate the expressing levels of signal transducer and activator of transcription genes (STAT1 and STAT2) and IFN-alpha stimulated genes (ISGs) which may mediate the inhibitory effects of IFN-alpha on HCV. Firstly HCV replicon cell culture system was established by transfecting HCV replicon RNA transcribed in vitro into Huh7 cells and screening with G418. Secondly, the established HCV replicon cells were treated with various concentrations of IFN-alpha (0, 10, 25, 50, 100, 250, 500, 750, 1000, 2500, and 5000 IU/ml) for 72 h or treated with 1000 IU/ml of IFN-alpha for different lengths of time (0, 24, 48, 72, 96 h), then the levels of HCV RNA and NS5A protein in these cells were examined by semi-quantitative RT-PCR and Western blot respectively. IFN-alpha could effectively inhibit HCV RNA replication. 10 IU/ml or 25 IU/ml of IFN-alpha could lead to about 68% and 75% of HCV-RNA reduction respectively. The cells treated with 1000 IU/ml IFN-alpha for 24 h or 96 h had about 75% and 88% of HCV RNA reduction compared with the cells of control, demonstrated that the inhibitory effects of IFN-alpha on HCV replicon are in dose and time dependent manners. The expressions of antiviral ISGs-PKR, 2'5'OAS, G1P3, ISG20 and ISGF3gamma are strongly induced by IFN-alpha. HCV replicons were sensitive to IFN-alpha treatment. The inhibitory effects of IFN-alpha on HCV RNA and NS5A are both dose and time dependents. PKR, 2'5'OAS, G1P3, ISG20 and ISGF3gamma may mediate the inhibitory effects of IFN-alpha on HCV replicon replication.


[Patient- and hospital-related determinants of satisfaction with hospital stay of severely injured patients].
General political and legal changes in the German health care system will increase the importance of patient surveys. Patient satisfaction with medical and organizational care during their stay in a hospital is a relevant outcome of quality assurance. Satisfied patients also show a higher compliance with treatment and therefore increase the probability of a successful outcome. The objective of this analysis was to identify determinants of satisfaction with hospital stay of severely injured patients. A written questionnaire was sent to 121 severely injured patients, who were predominantly hurt by traffic or work-related accidents and were treated between 1996 and 2001 in two hospitals in Cologne (response rate=74.4%, n=90). The questionnaire used in this study contained questions about psychosocial care of physicians and nurses in and organizational and structural characteristics of the hospital. Logistic regressions identified two significant predictors for satisfaction with hospital stay: (1) subjective evaluation of psychosocial quality of care by physicians and (2) patients' perception of active integration in treatment decisions. The results of this analysis confirm the importance of soft science in medical education also for surgeons. Medical education should integrate knowledge about psychosocial aspects of medicine. The quality of psychosocial care of physicians seems to be a predictor of patient satisfaction. The probability of dissatisfaction increases with deficits in communication, information, and time as well as no active integration in treatment. The results of this analysis confirm the vital importance of psychosocial aspects in physician-patient interactions with severely injured patients for satisfaction with stay in hospital.


Uptake and reduction of [N]nitrate by intact soybean plants in the dark.
Experiments were conducted to determine if nitrate ((15)N-labeled) was taken up and assimilated by intact soybean (Glycine max [L.] Merr. cv Williams) plants during extended periods of dark. Nitrate was taken up by soybean roots throughout a 12-hour dark period. The (15)N-labeled nitrogen was also translocated to the plant shoots, but at a slower rate than the rate of accumulation in the roots. Much of the nitrogen ((15)N-labeled) was present in a nonreduced form, although considerable soluble-reduced nitrogen ((15)N-labeled) accumulated throughout the dark period. The (15)N-labeled, soluble-reduced nitrogen fraction accounted for nearly 30% of the total (15)N found in plant roots and more than 63% of the total (15)N found in plant tops after 12 hours of dark. This provided evidence that intact soybean plants take up and metabolize significant quantities of nitrate to reduced N forms in the dark.In addition to nitrate influx during the dark, it was shown that there was a concomitant loss of (15)N-labeled nitrogen compounds from previously (15)N-labeled plants to a natural abundance (15)N nutrient solution. Thus, evidence was obtained which indicated that light was not directly essential for flux and reduction of nitrate by intact soybean plants.


Staging dementia using Clinical Dementia Rating Scale Sum of Boxes scores: a Texas Alzheimer's research consortium study.
The Clinical Dementia Rating Scale Sum of Boxes (CDR-SOB) score is commonly used, although the utility regarding this score in staging dementia severity is not well established. To investigate the effectiveness of CDR-SOB scores in staging dementia severity compared with the global CDR score. Retrospective study. Texas Alzheimer's Research Consortium minimum data set cohort. A total of 1577 participants (110 controls, 202 patients with mild cognitive impairment, and 1265 patients with probable Alzheimer disease) were available for analysis. Receiver operating characteristic curves were generated from a derivation sample to determine optimal cutoff scores and ranges, which were then applied to the validation sample. Optimal ranges of CDR-SOB scores corresponding to the global CDR scores were 0.5 to 4.0 for a global score of 0.5, 4.5 to 9.0 for a global score of 1.0, 9.5 to 15.5 for a global score of 2.0, and 16.0 to 18.0 for a global score of 3.0. When applied to the validation sample, kappa scores ranged from 0.86 to 0.94 (P < .001 for all), with 93.0% of the participants falling within the new staging categories. The CDR-SOB score compares well with the global CDR score for dementia staging. Owing to the increased range of values, the CDR-SOB score offers several advantages over the global score, including increased utility in tracking changes within and between stages of dementia severity. Interpretive guidelines for CDR-SOB scores are provided.


Root surface roughness following Er:YAG laser irradiation at different radiation energies and working tip angulations.
The determination of roughness of root surfaces following treatment with 2.94 micro m Er:YAG laser radiation at different radiation energies and working tip angulations. The study sample comprised 85 extracted human molars, premolars, canines and incisors (n = 85). The source of laser radiation was an Er:YAG laser device (KAVO-Key II, System Aesculap Meditec) emitting pulsed infrared radiation at a wavelength of 2.94 micro m, with a pulse duration of 250 micro s, and a pulse repetition rate of 10 pps. The samples were randomly divided into three experimental units, for treatment with a constant amount of 380 laser pulses at a radiation energy of 60 mJ, 100 mJ, and 180 mJ. Each experimental unit was divided into five subgroups of five samples, which were irradiated at a working tip angulations of 15 degrees, 30 degrees, 45 degrees, 60 degrees, and 90 degrees. Five samples were treated mechanically using curettes. Five samples were left untreated as control. The mean (Ra) and maximum (Rmax) surface roughness of each sample was measured using a profilometer. The statistical analysis was undertaken using anova and Scheffé-test at a level of significance of 5% (p < 0.05). Er:YAG laser radiation led to an Ra which ranged from 0.52 micro m (+/- 0.10) to 0.81 micro m (+/- 0.26) and to an Rmax between 3.4 micro m (+/- 0.48) and 9.26 micro m (+/- 3.08). The Ra and Rmax for samples treated with curettes was 0.51 micro m (+/- 0.11) and 5.08 micro m (+/- 4.98), respectively. That for the untreated control samples were 0.53 micro m (+/- 0.15) and 7.07 micro m (+/- 5.48), respectively. The mean and maximum surface roughness of root surfaces following irradiation with Er:YAG laser was not significantly different to that obtained on samples treated with conventional hand instruments or left untreated. Furthermore, the surface roughness does not depend on the radiation energy and the angulation of the working tip.


Occupational exposure of animal caretakers to cyclophosphamide.
Little is known about the exposure of animal caretakers to toxic agents during the administration of such chemicals to laboratory animals. In this study, we have investigated the environmental contamination with cyclophosphamide (CP) in an animal laboratory where mice were housed and injected with this compound. Also the contamination of gloves, sleeve protectors, and masks used for personal protection was studied. The uptake of CP by the animal caretakers was determined by the analysis of unmetabolized CP in urine. For the estimation of CP in the air, air samples were taken and filters of the air-circulation system were analyzed. On the filters, amounts of CP were detected corresponding with < 0.1-1.0 microgram/day. Environmental contamination was also measured by analysis of wipe samples taken from different spots (objects and surfaces). The presence of CP was not only observed in the room where the mice were housed and treated with CP but also in adjacent rooms (< 0.02-44 ng/cm2). The gloves used during the injection of CP were always contaminated (2-199 micrograms/pair). No penetration of the gloves was established. The sleeve protectors were incidentally contaminated (< 0.3-10 micrograms) and on the masks no CP was found (< 0.2 microgram). Eighty seven urine samples from four animal caretakers were analyzed for unmetabolized CP. In one sample, CP was detected (0.7 microgram). The results show that in this particular study animal caretakers are exposed to CP during their work.


The effectiveness of three types of alternating pressure air mattresses in the prevention of pressure ulcers in Belgian hospitals.
To compare the effectiveness of multi-stage and one-stage alternating low-pressure air mattresses (ALPAM) and alternating pressure air mattress (APAM) overlays in preventing pressure ulcers among hospitalized patients, data were pooled (N = 617) from a study of patients allocated to multi-stage ALPAM (n = 252) or one-stage ALPAM (n = 264), and another study of patients allocated to APAM overlay (n = 101). Cumulative pressure ulcer incidence was 4.9% (n = 30) over 14 days. Fewer ulcers developed on multi-stage ALPAM compared with APAM overlay (OR = 0.33; 95% CI [0.11, 0.97]), but no difference was found between one-stage ALPAM and APAM overlay (OR = 0.40; 95% CI [0.14, 1.10]). Time to develop ulcers did not differ by mattress type.


Clinical characteristics, surgical strategies, and outcome of solid pseudopapillary tumor of the pancreas: retrospective analysis in a single center.
Solid pseudopapillary tumor of the pancreas (SPTP) is a rarely diagnosed, low-malignancy pancreatic neoplasm, which mostly can be cured by surgery. To investigate the surgical effect and prognosis of SPTP. The data of 39 patients diagnosed with SPTP and treated with surgery between 2013 and 2020 were analyzed retrospectively. The data included the clinical characteristics, surgical management, pathological findings and therapeutic outcome. The mean age of the patients was 34.0 ±12.1 years, and the female : male ratio was 32 : 7. Most of the patients were asymptomatic (48.7%). The mean diameter of the tumors was 4.81 ±2.36 cm. Operative procedures were conducted according to the location and size of the tumors. Laparoscopic surgery, especially laparoscopic distal pancreatectomy (LDP), provided a smaller incision, a shorter postoperative hospital stay and a shorter postoperative fasting time. There was no observed difference in the amount of blood loss or complication rate. The median follow-up was 24 months. One patient with 20% expression of Ki-67 developed liver metastasis after surgery. SPTP is a rare disease with low malignancy. Minimally invasive surgery, especially LDP, has been proven to be a feasible and safe treatment method for SPTP with early recovery. The prognosis of SPTP is favorable. Lifetime surveillance is necessary especially in patients with a high expression rate of Ki-67.


Elevated expression of activation molecules by decidual lymphocytes in women suffering spontaneous early pregnancy loss.
The purpose of this study was to investigate, quantify and compare the expression of activation markers by decidual leukocytes in sporadic spontaneous early pregnancy loss and apparently normal first trimester human pregnancy. Decidua was obtained from 18 therapeutic abortions and 20 sporadic spontaneous abortions at 8-12 weeks gestational age. Cryostat sections were labelled by the avidin-biotin complex-peroxidase method using monoclonal antibodies specific for CD45, CD56, CD3, human leukocyte antigen (HLA) DR, CD69, CD25 and very late antigen (VLA)1. Positive cells were quantified and the results were analysed using the Mann-Whitney statistical test. Significantly increased numbers of CD69-positive and CD25-positive cells were detected in spontaneous abortion decidua, when compared with therapeutic abortion decidua. Approximately 50% of women experiencing spontaneous miscarriage also contained significantly elevated numbers of HLA DR-positive cells within decidua. Double immunohistochemical labelling studies demonstrated that the CD25-positive and CD69-positive cells in spontaneous abortion decidua were CD3-positive T cells rather than CD56-positive granulated lymphocytes. Immunological dysfunction within endometrium may account for a proportion of sporadic spontaneous abortions.


Gamma secretase inhibitor enhances sensitivity to doxorubicin in MDA-MB-231 cells.
Deregulated expression of molecular of the Notch signaling pathway is observed in malignant tumor. Notch signaling pathway is activated by a series of catalytic cleavage of the Notch receptor by gamma secretase. Gamma secretase inhibitor (GSI) have demonstrated clinical activity in patients with solid tumor. Triple negative breast cancer (TNBC) is related to poor prognosis and a high probability of lung and brain metastases. As first line therapy for TNBC, doxorubicin is partially effective in TNBC control. An understanding of the mechanisms for enhancing sensitivity to doxorubicin would be significant for future drug development. We hypothesized that a combination of cytotoxic chemotherapy doxorubicin to inhibit cell proliferation, together with GSI, would result in more effective outcome than either monotherapy alone. We treated MDA-MB-231 cell lines with doxorubicin and evaluated the monotherapy efficacy and in combination with GSI in both vitro and vivo. GSI-induced proliferation inhibition and apoptosis was achieved with an induction of PTEN and pro-apoptotic protein Bax expression and suppression of Notch-1, HES-1, CyclinD1 and anti-apoptotic protein Bcl-2. These results indicate that MDA-MB-231 cells are susceptible to a GSI, whether alone or in combination with doxorubicin, are correlated with changing of some surrogate marker. This study demonstrates practicability of combined use of GSI and doxorubicin, and together these results encourage new therapeutic method in triple negative breast cancer.


Recombinant calpain II: improved expression systems and production of a C105A active-site mutant for crystallography.
The bacterial production of recombinant rat calpain II has been improved greatly by the use of two compatible plasmids for the two subunits. The calpain small subunit C-terminal fragment (21 kDa) was expressed from a new A15-based vector created by cloning T7 control elements into pACYC177. This vector is compatible with the ColE1-based pET-24d(+) vector containing the calpain large subunit, and the yield of calpain activity was increased at least 16-fold by co-expression from these two vectors. A high level of activity was also obtained from a bicistronic construct containing both subunit cDNAs under the control of one T7 promoter. The addition of a C-terminal His-tag to the large subunit simplified purification without affecting subunit association or enzyme activity. The active-site cysteine 105 was mutated to alanine, causing complete loss of activity. The yield of purified C105A-calpain II (80 + 21 kDa) dimer following three column chromatography steps was 10 mg/l of cell culture. This provides a purified calpain, stable to autolysis and oxidation, which is likely to facilitate crystallization in both the presence and absence of calcium.


Assembly and Application of a Three-Dimensional Human Corneal Tissue Model.
The cornea provides a functional barrier separating the outside environment from the intraocular environment, thereby protecting posterior segments of the eye from infection and damage. Pathological changes that compromise the structure or integrity of the cornea may occur as a result of injury or disease and can lead to debilitating effects on visual acuity. Over 10 million people worldwide are visually impaired or blind due to corneal opacity. Thus, physiologically relevant in vitro approaches to predict corneal toxicity of chemicals or effective treatments for disease prior to ocular exposure, as well as to study the corneal effects of systemic, chronic conditions, such as diabetes, are needed to reduce use of animal testing and accelerate therapeutic development. We have previously bioengineered an innervated corneal tissue model using silk protein scaffolds to recapitulate the structural and mechanical elements of the anterior cornea and to model the functional aspects of corneal sensation with the inclusion of epithelial, stromal, and neural components. The purpose of this unit is to provide a step-by-step guide for preparation, assembly, and application of this three-dimensional corneal tissue system to enable the study of corneal tissue biology. © 2019 by John Wiley & Sons, Inc.


Spanish version of Bus Drivers' Job Demands Scale (BDJD-24).
Karasek and Theorell's Job Demands-Control Model argues that adverse health-related outcomes, both psychological and physiological, arise from a combination of high job demand and a low level of job control. The objective was to adapt Meijman and Kompier's Bus Drivers' Job Demands Scale (BDJD-24), which enables us to assess the job demands of bus drivers, to Spanish. The final version of the Spanish adaptation was applied to a sample made up of 287 bus drivers living in Spain (80.1% men and 19.9% women), whose average age was 40.44 (SD= 11.78). The results yielded a three-factor structure for the scale used: Time Pressure, Safety, and Passengers. These findings confirm that the Spanish version replicates the factor structure of the original English scale. The reliability of the three subscales was acceptable, ranging from .75 to .84. Furthermore, the subscales were also related to different external correlates and to other scales and showed good convergent and criterion validity. The present instrument can be used to evaluate job demands of bus drivers, as its psychometrics are substantially sound.


Identifying and reducing AFLP genotyping error: an example of tradeoffs when comparing population structure in broadcast spawning versus brooding oysters.
Phylogeographic inferences about gene flow are strengthened through comparison of co-distributed taxa, but also depend on adequate genomic sampling. Amplified fragment length polymorphisms (AFLPs) provide a rapid and inexpensive source of multilocus allele frequency data for making genomically robust inferences. Every AFLP study initially generates markers with a range of locus-specific genotyping error rates and applies criteria to select a subset for analysis. However, there has been very little empirical evaluation of the best tradeoff between culling all but the lowest-error loci to minimize overall genotyping error versus the potential for increasing population genetic signal by retaining more loci. Here, we used AFLPs to compare population structure in co-distributed broadcast spawning (Crassostrea virginica) and brooding (Ostrea equestris) oyster species. Using existing methods for almost entirely automated marker selection and scoring, genotyping error tradeoffs were evaluated by comparing results across a nested series of data sets with mean mismatch errors of 0, 1, 2, 3, 4 and >4%. Artifactual population structure was diagnosed in high-error data sets and we assessed the low-error point at which expected population substructure signal was lost. In both species, we identified substructure patterns deemed to be inaccurate at average mismatch error rates 2 and >4%. In the species comparison, the optimum data sets showed higher gene flow for the brooding oyster with more oceanic salinity tolerances. AFLP tradeoffs may differ among studies, but our results suggest that important signal may be lost in the pursuit of 'acceptable' error levels and our procedures provide a general method for empirically exploring these tradeoffs.


ATF3 modulates calcium signaling in osteoclast differentiation and activity by associating with c-Fos and NFATc1 proteins.
Activating transcription factor 3 (ATF3), a member of the ATF/cAMP response element-binding protein family of transcription factors, has been implicated in the regulation of cell proliferation and differentiation. However, whether ATF3 is involved in osteoclast differentiation and activity has not been well-studied. In the present study, we examined the role of ATF3 in osteoclast differentiation and function. ATF3 expression was down-regulated during RANKL-induced osteoclast differentiation. Overexpression of ATF3 in bone marrow-derived monocyte/macrophage lineage cells (BMMs) promoted osteoclast differentiation and activity and strongly induced the expression of osteoclast genes encoding nuclear factor of activated T-cells c1 (NFATc1) and tartrate-resistant acid phosphatase (TRAP) compared to that in the control group. In contrast, small interfering RNA-mediated knockdown of ATF3 prevented the formation of multinucleated osteoclasts and markedly abrogated the expression of osteoclast marker genes. Mechanistically, ATF3 synergistically enhanced c-Fos- or NFAT-mediated transcriptional activity of the NFATc1 or TRAP promoter, respectively. Furthermore, ATF3 physically interacted with c-Fos and NFATc1 and enhanced the binding affinity of c-Fos and NFATc1 to the promoters. Interestingly, ATF3 is involved in calcium signaling during osteoclastogenesis. Taken together, these results suggest that ATF3 is a new co-factor of c-Fos and NFATc1 to activate osteoclast differentiation and activity.


Effects of a 10-Strain Oral Probiotic on Parameters of Vaginal Health and Microbial Community: A Pilot Clinical Study.
To investigate the clinical effects of a 10-strain probiotic on parameters of vaginal health in a pilot, open label study in women with intermediate Nugent score (NS) or vaginal pH >4.5. A total of 43 healthy premenopausal women, ages 18 to 50 years, with NS of 4-6 or vaginal pH >4.5 were enrolled. Participants consumed a probiotic formulation (Feminine Support™), containing 8 lactobacilli and 2 bifidobacteria strains, with a daily dose of 2.5×1010 CFU for 28 (subgroup 1) or 42 (subgroup 2) days. Investigational visits occurred at day 0, 14, 28 and 42 with assessment of vaginal pH, NS and vaginal microbiota, via next-generation sequencing. A total of 36 participants were included in the analysis set, with 24 and 12 participants included in subgroups 1 and 2, respectively. In the analysis set, there was a significant reduction in vaginal pH, from baseline, at day 28 (mean change=-0.19, P = 0.047). Participants in subgroup 1 achieved a significant reduction in vaginal pH from baseline, at day 28 (mean change=-0.23, P = 0.029) and day 42 (mean change=-0.29, P = 0.008), while participants in subgroup 2 achieved a significant and quantitatively greater reduction in vaginal pH from baseline to day 42 (mean change=-0.64, P = 0.008). No significant changes in NS were reported, due in part to highly diverse baseline levels. Vaginal microbial abundance exhibited a majority lactobacilli abundance at baseline, which was maintained over the study period. Vaginal pH was inversely associated with lactobacilli abundance throughout the study (P < 0.005). The product was well tolerated with high compliance. Two participants reported adverse events with suspected causality, which were mild and resolved during the study. This 10-strain probiotic formulation was well tolerated and helped reduce vaginal pH in women with intermediate NS or elevated vaginal pH. The study product warrants a randomized controlled trial to further assess efficacy.


Graphene-embedded first-order mode polymer Mach-Zender interferometer thermo-optic switch with low power consumption.
Two-dimensional-material integrated thermal optical switches have low power consumption; however, the devices are suffering from high propagation loss due to the two-dimensional material absorption. In this Letter, we present a graphene-embedded polymer Mach-Zender interferometer (MZI) thermo-optic switch, based on the E01x mode and designed to minimize both the loss and power consumption. Based on the symmetry of a three-dimensional structure and the E01x mode, the central embedded graphene electrode structure was simulated with an absorption loss of 0.06 dB/cm. Finite element method (FEM) simulations were run to find that the power consumption is 1.57 mW. Compared with the top heating electrode, the power consumption of the proposed graphene-embedded device is reduced by 74%. Further, the response speed of the graphene-embedded thermo-optic MZI switch is simulated to be 1.2 μs (rise) and 70.6 μs (down). This device may be applicable in the two-dimensional integrated low-power-consumption-mode division multiplexer field.


Identification of two independent transcriptional activation domains in the Autographa californica multicapsid nuclear polyhedrosis virus IE1 protein.
The Autographa californica multicapsid nuclear polyhedrosis virus immediate-early protein, IE1, is a 582-amino-acid phosphoprotein that regulates the transcription of early viral genes. Deletion of N-terminal regions of IE1 in previous studies (G. R. Kovacs, J. Choi, L. A. Guarino, and M. D. Summers, J. Virol. 66:7429-7437, 1992) resulted in the loss of transcriptional activation, suggesting that this region may contain an acidic activation domain. To identify independently functional transcriptional activation domains, we developed a heterologous system in which potential regulatory domains were fused with a modified Escherichia coli Lac repressor protein that contains a nuclear localization signal (NLacR). Transcriptional activation by the resulting NLacR-IE1 chimeras was measured with a basal baculovirus early promoter containing optimized Lac repressor binding sites (lac operators). Chimeras containing IE1 peptides dramatically activated transcription of the basal promoter only when lac operator sequences were present. In addition, transcriptional activation by NLacR-IE1 chimeras was allosterically regulated by the lactose analog, isopropyl-beta-D-thiogalactopyranoside (IPTG). For a more detailed analysis of IE1 regulatory domains, the M1 to T266 N-terminal portion of IE1 was subdivided (on the basis of average amino acid charge) into five smaller regions which were fused in various combinations to NLacR. Regions M1 to N125 and A168 to G222 were identified as independent transcriptional activation domains. Some NLacR-IE1 chimeras exhibited retarded migration in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels. As with wild-type IE1, this aberrant gel mobility was associated with phosphorylation. Mapping studies with the NLacR-IE1 chimeras indicate that the M1 to A168 region of IE1 is necessary for this phosphorylation-associated effect.


Efficacy and safety of embolization in arteriovenous malformations of the extremities and head and neck: a retrospective study of 32 cases.
Definitive treatment of arteriovenous malformations (AVMs) consists of complete surgical excision. When not possible, embolization may be performed. We aimed to evaluate the efficacy and safety of embolization for AVMs of the extremities and head and neck. This retrospective study included all patients undergoing embolization for AVMs of skin and soft tissues on the limbs, head and neck, in the University Hospital Center of Tours between January 1996 and December 2009. The main outcome was efficacy, assessed by two independent assessors, as the percentage of AVM devascularized at the end of embolization. Secondary outcomes were patient satisfaction, evolution of symptoms and safety of embolization. We included 32 AVMs in 31 patients, for 66 embolizations evaluated. In 18 AVMs (56.3%), devascularization was greater than 75% of the initial vascularization. Efficacy was lower for AVMs of the upper limbs than other body parts (p = 0.003). For 18 patients who could be contacted by telephone, the mean global satisfaction was 6.0 ± 2.7 on a scale of 0 to 10, and 13 (72.2%) reported an improvement of the symptoms linked to the AVM. Severe adverse events were reported after 4 embolizations, all located on the head and neck. Embolization can lead to good devascularization and improve symptoms linked to AVMs, especially in lower limbs. Minor complications are frequent, and severe adverse events may occur, especially after procedures on the head and neck.


Painful pulp space of a pinky finger: A report of glomus tumor at an unusual site.
The long-standing pain in fingers is not only agonising and disabling to the patient but also affects the quality of life. Pointed fingertip pain is also inappropriately managed for years leading to further morbidity. Pinpoint tenderness, paroxysmal pain, and cold hypersensitivity at finger pulp space may be features of glomus tumor. Judicious use of investigations and surgical excision ensure an optimal outcome. We report a case of glomus tumor at an unusual location at the pulp space of distal phalanx of the little finger that was managed by en bloc removal and confirmed on biopsy. History of 6 years of clinical symptoms and multiple treatments underlines the importance of the knowledge of this lesion for early diagnosis.


Hepcidin-Dependent Regulation of Erythropoiesis during Anemia in a Teleost Fish, Dicentrarchus labrax.
Anemia is a common disorder, characterized by abnormally low levels of red blood cells or hemoglobin. The mechanisms of anemia development and response have been thoroughly studied in mammals, but little is known in other vertebrates, particularly teleost fish. In this study, different degrees of anemia were induced in healthy European sea bass specimens (Dicentrarchus labrax) and at pre-determined time points hematological parameters, liver iron content and the expression of genes involved in iron homeostasis and hematopoiesis, with particular attention on hepcidins, were evaluated. The experimental anemia prompted a decrease in hamp1 expression in all tested organs, in accordance to an increased need for iron absorption and mobilization, with slight increases in hamp2 in the kidney and intestine. The liver was clearly the major organ involved in iron homeostasis, decreasing its iron content and showing a gene expression profile consistent with an increased iron release and mobilization. Although both the spleen and head kidney are involved in erythropoiesis, the spleen was found to assume a more preponderant role in the recovery of erythrocyte levels. The intestine was also involved in the response to anemia, through the increase of iron transporting genes. Administration of Hamp1 or Hamp2 mature peptides showed that only Hamp1 affects hematological parameters and liver iron content. In conclusion, the molecular mechanisms of response to anemia present in sea bass are similar to the ones described for mammals, with these results indicating that the two hepcidin types from teleosts assume different roles during anemia.


[Minimycetomas caused by N. brasiliensis. Histological study of 13 cases].
We present a preliminary communication of the histological findings in 16 cases of mini-mycetoma by N. brasiliensis. We found little PAS positive granules, fully evident with the Gram stain, localized at the center of the microabscesses. Ten cases showed Russell bodies, 33% showed multinucleated giant cells and the 50%, epithelioid cells. All cases showed abundant capillary vessels and intense fibrosis.


[Nitric oxide--basal biochemistry and physiological aspects].
Nitric oxide (NO) is a naturally occurring molecule found in a variety of cell types and organ systems, including the cardiovascular, immune and nervous system. NO is normally produced in the endothelium from L-arginine by the constitutive isoform of the NO synthase (cNOS). Thereby, NO is an important regulator of vascular tone, prevents platelet adhesion, aggregation and activation, limits leukocyte adhesion to the endothelium and regulates myocardial contractility. This physiological production of NO is important for blood pressure regulation, blood flow distribution and tissue perfusion. Following injury or certain inflammatory stimuli, the expression of an inducible NO synthase (iNOS) can occur in a great variety of cells. In the last decade research on NO has suggested new treatment strategies for several diseases. In this review we discuss the biochemistry of NO and its basal physiological implications, with special emphasis on NO produced in the endothelium.


Impaction collection and slurry sampling for the determination of arsenic, cadmium, and lead in sidestream cigarette smoke by inductively coupled plasma-mass spectrometry.
The slurry sampling technique has been applied for the determination of arsenic, cadmium and lead in sidestream cigarette smoke condensate (SS CSC) by inductively coupled plasma mass spectrometry (ICP-MS). The smoke collection system consists of a "fishtail" chimney in conjunction with an impaction trap and a mixed cellulose ester (MCE) backup filter for the SS CSC collection. The smoke condensates, collected at three different regions of the system (i.e. the inner wall of the chimney, the impaction trap and the MCE filter), were individually prepared and analyzed and showed that about 70 and 20% of Cd and Pb, respectively, were trapped by the impaction trap and the MCE filter with less than 5% trapped by the chimney. In contrast, about 60% of As was trapped by the impactor and a relatively high percentage of As, about 30%, was deposited onto the inner wall of the chimney. The uneven distribution of trace metals at each collection stage strongly suggested that the composition of smoke aerosol and the associated physical form of the analytes may be different. The yields of As, Cd and Pb for the sidestream cigarette smoke of the 1R4F reference cigarette were 29.5 +/- 2.4, 421 +/- 6.6 and 46.4 +/- 0.9 ng cigarette(-1)(n= 3) respectively, when the cigarette was smoked according to the smoking regime: 45 mL puff volume of 2-s puff duration at an interval of 30 s with 50% ventilation holes covered. The analytical results for the slurry sampling technique were also compared with other sample preparation techniques and show good agreement.


The wrath of grapes versus the self-medication hypothesis.
Dr. Khantzian's SMH was initially developed to enhance an etiologic understanding of substance use disorders and to promote appropriate and empathic therapeutic responses through careful listening to the human suffering of addicted patients. In the present review he has combined an intensive look at patients with a full exploration of the modern literature on comorbidity and a discussion of the ways in which this more recent work may support or contradict the SMH. Additional longitudinal research is needed to confirm and delineate the SMH. There is a danger that clinicians could use the SMH in overly reductionistic ways. They must be alert to potential rationalization and retrospective distortion while listening to their patients' accounts of the causes of their substance abuse. In the cart/horse problem, it can be clinically useful to tease apart the temporal sequence of onset of each disorder, the ways in which the two disorders interact, and the ways in which each may amplify the suffering caused by the other. In my view, it is still the wrath of grapes that leads to the greater part of suffering in substance use disorders.


Parasitic Hematuria: Six Cases in a Row in a Single Centre in Spain.
Most cases of urogenital parasitosis are registered in Africa. However, migration movements and travellers moving from developed to developing countries are responsible for leading to an increased incidence of genitourinary infections caused by parasites in the western world including Spain having serious economic and health implications. The importance of its early detection and treatment also results from its potential risk for infertility, susceptibility for HIV infection and the development of bladder cancer. The most common presentation symptom is terminal haematuria, and when diagnosed, praziquantel is the treatment of choice. We report a series of 6 cases of urinary schistosomiasis that happened in a single centre in Spain and reminds the importance of having the infection in mind in certain cases of haematuria study.


Sensori-motor function in older persons with diabetes.
Twenty-five persons with diabetes (aged 55-83 years) who were living independently in the community, and 40 age- and sex-matched non-diabetic controls were assessed for tactile sensitivity, vibration sense, proprioception, quadriceps strength and body sway. In both men and women, those with diabetes performed significantly worse in tests of body sway on firm and compliant surfaces compared with the control subjects after controlling for weight and body mass index. The female diabetic subjects also performed significantly worse in tests of peripheral sensation and strength compared with controls. Age-related declines in sensori-motor function were greater in the diabetic group (r = 0.55-0.75) than in the controls (r < 0.44), while within the diabetic group, duration of diabetes and vibration sense were significantly correlated with sway on a compliant (foam rubber) surface with the eyes open (partial r = 0.52, p < 0.01 and r = 0.55, p < 0.01, respectively). The study findings provide evidence that older people with diabetes have problems with stability and related sensori-motor factors which may place them at increased risk of falls.


Laparoscopic sleeve gastrectomy reduces the predicted coronary heart disease risk and the vascular age in obese subjects.
Obesity is associated with high prevalence of coronary heart disease (CHD) and long term increased cardiovascular morbi-mortality. There are no data regarding the effect of laparoscopic sleeve gastrectomy (LSG) on long-term CHD - risk. It is known that "a man is as old as his arteries" and this concept is illustrated by Framingham coronary risk score, which can predict vascular age. To assess the 10-year CHD risk in patients with obesity, preoperatively, and 6 and 12 months after LSG. 47 consecutive obese subjects (44.7% males, mean age 39.8 years) scheduled for LSG were prospectively studied before and 6 and 12 months after surgery. The 10 years CHD risk and corresponding vascular age were calculated using Framingham risk score. The body mass index (BMI) decreased from 44.6 ± 10.6 kg m2 preoperatively to 32.2 ± 6.9 kg m2 and to 29.4 ± 5.4 kg m2 at 6 and 12 months follow-up (both p 0.05). Mean excessive weight loss (EWL) was 67.3 ± 23.7% and 78.3 ± 23.4% at 6 and 12 months postoperatively. At 6 and 12 months after LSG, there was a marked improvment of lipid profile(decrease of total cholesterol, LDL-cholesterol, triglycerides and increase of HDL-cholesterol) and a significant decrease in prevalence of diabetes mellitus, systemic hypertension and smoking. The 10-year CHD risk reduced from 10.1% preoperatively to 3.5% and to 2.2% at 6 and 12 months after surgery (both p 0.05). Patients' mean vascular age was 65.6 years preoperatively and decreased to 45.8 years 6 month spostoperatively (p 0.05) and to 40.7 years one year after LSG (p 0.05 vs. 6 months postoperatively, p=NS vs.chronological age). In obese subjects, CHD risk is significantly reduced early, beginning with 6 months after LSG and is diminished with 80% one year postoperatively. Despite the fact that not all patients had achieved the ideal weight yet,mean vascular age is similar to their chronological age one year after surgery.


Recent advances in head and neck free tissue transfer.
Free tissue transfer is a versatile and valuable method for reconstructing select head and neck defects following trauma or oncologic resection. Microvascular reconstructive cases are among the longest, most technically challenging, and most labor-intensive operations performed by departments of otolaryngology/head and neck surgery. However, technical advances, increased experience, and robust training programs have permitted realization of microvascular success rates in excess of 97% at most high-volume centers. Given this unprecedented degree of success, research emphasis has shifted to advancing techniques, expanding indications, and increasing efficiencies. Although numerous topics are important for discussion, this update focuses on recent notable advances in reconstruction. These include expanding utility of the anterolateral thigh free flap in soft tissue reconstruction, prefabricated plating for fibula free flap mandibular reconstruction, use of venous couplers, and postoperative free tissue monitoring techniques. Improvements in technique, technology, and monitoring continue to improve success rates, reduce operative time and associated morbidity, improve overall functional outcomes, and improve patient-specific quality of life. These highlighted recent advances, amongst others, promote further advancement and simplification of reconstructive capabilities.


Hyping Hypnosis: The Myth That Made Capturing the Friedmans Persuasive.
Those promoting the idea of "false memory syndrome" often invoke the specter of hypnosis to discredit those making accusations of sexual abuse and anyone they might have spoken to for investigative or therapeutic purposes. Capturing the Friedmans demonstrates that accusations of hypnosis have strong rhetorical value, even when they are not true. The film, classified as a documentary, tells the story of a family that is shattered when the father and son both plead guilty to sexually abusing boys in the after-school classes in their basement. Using tropes about hypnosis and misrepresenting the actual facts in the case, the movie persuaded many people that Jesse Friedman was actually innocent. A detailed Conviction Integrity Review that was prompted by the movie demonstrates that the conviction was sound, and that the movie is suspect.


Analysis of abnormalities of capillary CO2 exchange in vivo.
Capillary CO2 exchange in vivo is affected by several interdependent reactions and transport processes. A mathematical model that includes all the significant chemical and transport events that are presumed to occur during capillary gas exchange has been used to investigate the effect of inhibition of 1) erythrocyte HCO(3-)-Cl- exchange, 2) lung carbonic anhydrase (CA) activity with access to plasma, and 3) erythrocyte CA activity on overall pulmonary CO2 excretion (VCO2) during rest and moderate exercise. Any decrement in VCO2 due to inhibition of HCO(3-)-Cl- exchange and/or CA activity, should result in compensatory alterations in cardiac output and/or an increase in the mixed venous blood-to-alveolar PCO2 gradient [(delta PCO2)V-A] to restore steady-state VCO2. Our computations show that complete inhibition of erythrocyte anion exchange would require a compensatory increment in cardiac output of approximately 30-40% or an increase in (delta PCO2)V-A from 6 to 8.3 Torr at rest and from 12 to 15.6 Torr during moderate exercise, if lung CA activity is intact. In the absence of availability of lung CA activity to plasma, the necessary (delta PCO2)V-A is 10.5 Torr at rest and 19.5 Torr during moderate exercise. Complete inhibition of lung and erythrocyte CA activity is predicted to require (delta PCO2)V-A of 39.1 Torr at rest and 74.2 Torr during moderate exercise. These results suggest that HCO(3-)-Cl- exchange might not be vital to maintenance of CO2 transfer and perhaps has a more important role in minimizing the changes in plasma pH associated with microvascular gas exchange in vivo.


Serine protease 35 regulates the fibroblast matrisome in response to hyperosmotic stress.
Hyperosmotic stress occurs in several diseases, but its long-term effects are largely unknown. We used sorbitol-treated human fibroblasts in 3D culture to study the consequences of hyperosmotic stress in the skin. Sorbitol regulated many genes, which help cells cope with the stress condition. The most robustly regulated gene encodes serine protease 35 (PRSS35). Its regulation by hyperosmotic stress was dependent on the kinases p38 and JNK and the transcription factors NFAT5 and ATF2. We identified different collagens and collagen-associated proteins as putative PRSS35 binding partners. This is functionally important because PRSS35 affected the extracellular matrix proteome, which limited cell proliferation. The in vivo relevance of these findings is reflected by the coexpression of PRSS35 and its binding partners in human skin wounds, where hyperosmotic stress occurs as a consequence of excessive water loss. These results identify PRSS35 as a key regulator of the matrisome under hyperosmotic stress conditions.


Disorders linked to insufficient androgen action in male children.
Virilization of the external genitalia in the male fetus requires testosterone and dihydrotestosterone (DHT), which is formed from testosterone by the action of the enzyme, 5alpha-reductase type 2 (5alphaR-2). Mediation of the effects of both testosterone and DHT requires a functional androgen receptor (AR) located in the cytoplasmic compartment of target cells. DHT (or testosterone) binding induces a conformational change which facilitates AR nuclear transport, phosphorylation and dimerization, ultimately regulating of the rate of transcription of androgen-dependent genes. Any event which impairs DHT formation (mutation within the 5alphaR-2 gene or 5alphaR-2 inhibitors) or normal function of the AR (mutation in the AR gene, antiandrogens) may result in insufficient androgen action in the male fetus and in subsequent undervirilization in the newborn. Hypospadias may be due to a defect in androgen action due to mutation of the 5alphaR-2 or of the AR gene. Mutation of unidentified genes is likely to underlie this displacement of the urethral meatus from the tip to the ventral side of the phallus. An aetiological role for environmental chemical products has been postulated, since ethnic as well as geographical differences in the incidence of hypospadias have been noted. Increasing evidence has been gathered indicating that widely used industrial and agricultural chemicals have deleterious effects on normal male sexual differentiation. Cryptorchidism and micropenis may represent an intersex phenotype, even if they are isolated. Aetiological factors include 5alphaR-2 gene mutation, AR gene mutation or environmental hormonal disruptors. In conclusion, several phenotypes have been attributed to insufficient androgen action during fetal life. Whereas mutations in the 5alphaR-2 gene and AR gene are natural, attention should be focused on environmental endocrine disruptors that are able to mimic steroid 5alpha-reductase deficiency or partial androgen insensitivity syndrome.


Ligamentotaxis for Barton's and paediatric distal radial fractures.
To establish a consensus regarding immobilisation of the wrist following reduction of Barton's and paediatric distal radial fractures. Questionnaires were distributed to orthopaedic surgeons at the European Federation of National Associations of Orthopaedics and Traumatology meeting in Lisbon in 2005. Questions included the surgeon's country of practice, hospital, professional grade, years of experience, sub-specialty, and preferred position of wrist immobilisation after (1) a volar Barton's fracture, (2) a dorsal Barton's fracture, (3) a paediatric Salter-Harris type-II injury to the distal radius with volar displacement, and (4) the same injury but with dorsal displacement. Of 148 questionnaires distributed, 118 were returned. The specialist-to-trainee ratio was 45:73. In volar Barton's fractures, only 20% (29% specialists and 15% trainees) would immobilize the wrist in palmar flexion, as per recommendations. In dorsal Barton's fractures, only 25% (33% specialists and 21% trainees) would immobilize the wrist in dorsiflexion, as per recommendation. In paediatric Salter Harris type-II injury to the distal radius with volar displacement, 87% (100% specialists and 79% trainees) would immobilize the wrist in dorsiflexion or in a neutral position, as per recommendation. In the same injury but with dorsal displacement, 84% (89% specialists and 81% trainees) would immobilize the wrist in palmar flexion or in a neutral position, as per recommendation. In all 4 types of fractures, 26% to 30% of respondents would immobilize the wrist in a neutral position. Most respondents deviate from the recommended immobilisation positions in treating Barton's fractures. Understanding of the anatomy or biomechanics of ligamentotaxis are crucial for conservative treatments.


Physical fitness status modulates the inflammatory proteins in peripheral blood and circulating monocytes: role of PPAR-gamma.
The aim of this study was to analyze the metabolic and molecular profile according to physical fitness status (Low or High VO2max) and its impacts on peripheral and cellular inflammatory responses in healthy men. First (Phase I), inflammatory profile (TNF-α, IL-6, IL-10) was analyzed at baseline and post-acute exercise sessions performed at low (< 60% VO2max) and high (> 90% VO2max) intensities considering the individual endotoxin concentrations. Next (Phase II), monocyte cell cultures were treated with LPS alone or associated with Rosiglitazone (PPAR-γ agonist drug) to analyze cytokine production and gene expression. Monocyte subsets were also evaluated by flow cytometry. A positive relationship was observed between LPS concentrations and oxygen uptake (VO2max) (r = 0.368; p = 0.007); however, in the post-exercise an inverse correlation was found between LPS variation (Δ%) and VO2max (r = -0.385; p = 0.004). With the low-intensity exercise session, there was inverse correlation between LPS and IL-6 concentrations post-exercise (r = -0.505; p = 0.046) and a positive correlation with IL-10 in the recovery (1 h post) (r = 0.567; p = 0.011), whereas with the high-intensity exercise an inverse correlation was observed with IL-6 at pre-exercise (r = -0.621; p = 0.013) and recovery (r = -0.574; p = 0.016). When monocyte cells were treated with LPS, High VO2max individuals showed higher PPAR-γ gene expression whereas Low VO2max individuals displayed higher IL-10 production. Additionally, higher TLR-4, IKK1, and PGC-1α gene expression were observed in the High VO2max group than Low VO2max individuals. In conclusion, even with elevated endotoxemia, individuals with High VO2max exhibited higher IL-6 concentration in peripheral blood post-acute aerobic exercise and lower IL-10 concentration during recovery (1 h post-exercise). The anti-inflammatory effects linked with exercise training and physical fitness status may be explained by a greater gene expression of IKK1, TLR-4, and PGC-1α, displaying an extremely efficient cellular framework for the PPAR-γ responses.


Isolation of dominant Arabidopsis seiv mutants defective in VND7-induced xylem vessel cell differentiation.
The plant-specific NAC transcription factor VASCULAR-RELATED NAC-DOMAIN 7 (VND7) functions in xylem vessel cell differentiation in Arabidopsis thaliana. To identify novel factors regulating xylem vessel cell differentiation, we previously performed ethyl methanesulfonate mutagenesis of a transgenic 35S::VND7-VP16-GR line in which VND7 activity can be induced post-translationally by glucocorticoid treatment. We successfully isolated mutants that fail to form ectopic xylem vessel cells named seiv (suppressor of ectopic vessel cell differentiation induced by VND7) mutants. Here, we isolated eight novel dominant seiv mutants: seiv2 to seiv9. In these seiv mutants, ectopic xylem vessel cell differentiation was inhibited in aboveground but not underground tissues. Specifically, the shoot apices of the mutants, containing shoot apical meristems and leaf primordia, completely lacked ectopic xylem vessel cells. In these mutants, the VND7-induced upregulation of downstream genes was reduced, especially in shoots compared to roots. However, endogenous xylem vessel cell formation was not affected in the seiv mutants. Therefore, the seiv mutations identified in this study have repressive effects on cell differentiation in shoot meristematic regions, resulting in inhibited ectopic xylem vessel cell differentiation.


Heparin-binding epidermal growth factor-like growth factor/diphtheria toxin receptor in normal and neoplastic hematopoiesis.
Heparin-binding EGF-like growth factor (HB-EGF) belongs to the EGF family of growth factors. It is biologically active either as a molecule anchored to the membrane or as a soluble form released by proteolytic cleavage of the extracellular domain. HB-EGF is involved in relevant physiological and pathological processes spanning from proliferation and apoptosis to morphogenesis. We outline here the main activities of HB-EGF in connection with normal or neoplastic differentiative or proliferative events taking place primitively in the hematopoietic microenvironment.


[Anti-arrhythmic action of N-(aminoalkylene) derivatives of indoline-2,3-diones and 1,2-benzene-dicarboxiamides].
The aim of study was to modify the structure of indoline-2,3-diones and 1,2-benzenedicarboximides using the pharmacophores of new generation antiarrhythmics and to assess the impact of their structure upon the acute toxicity and antiarrhythmic action. The quaternary derivatives of N-aminoalkylindoline-2,3-diones and N-aminoalkyl-1,2-benzenedicarboximides were synthesized. The acute toxicity of compounds for white mice was tested. Using the calcium chloride- and aconitine-induced arrhythmia models in rats the antiarrhythmic action of derivatives was assessed. It was shown, that the antiarrhythmic action of N-aminoalkyl-1,2-benzenedicarboximides increases by lengthening of alkylene chain from one methylene group to two or by the presence of methanesulfonamide group in benzene ring. These structural changes, especially the presence of methanesulfonamido group, cause the decrease of acute toxicity. Among the N-aminoalkylindoline-2,3-diones the most antiarrhythmic action and minimal toxicity demonstrates the compound containing the bromo-substituted benzene ring.


Effects of orthodontic magnets on cutaneous epithelial thickness and tibial bone growth in rats.
The use of permanent magnets in orthodontic treatment has increased significantly over the past years, triggered by the introduction of rare earth magnetic alloys. Concerns about possible side effects have been expressed relating to their use in orthodontic treatment. In previous experimental studies evidence of a reduction in bone formation and epithelial turnover close to rare earth magnets has been presented. The aim of the present study was twofold: to confirm earlier results and to study whether the observed changes are reversible. One hind leg in each of 45 rats was fitted with aluminum rings with Co5Sm magnets, whereas the other hind leg was fitted with sham rings. After 8 weeks 25 rats were killed, and the rings in the remaining rats were removed. The latter group was killed after another 11 weeks. The epithelial and bone reactions were evaluated histomorphometrically. Previous results showing thinner epithelium and retarded rate of bone formation close to Co5Sm magnets were confirmed. These effects were also shown to be largely reversible. It was concluded on the basis of this and the magnitude of the effects that any local side effects from orthodontic rare earth magnets appear negligible when extrapolated to the clinical situation.


Basal Values of Biochemical and Hematological Parameters in Elite Athletes.
The purpose of this study was to show how continuous exercise affects the basal values of biochemical and hematological parameters in elite athletes. A total of 14,010 samples (male = 8452 and female = 5558 (March 2011-March 2020)) from 3588 elite athletes (male = 2258 and female = 1330, mean age 24.9 ± 6.9 vs. 24.1 ± 5.5 years, respectively) from 32 sport modalities, were studied over 9 years to check the variation of basal biochemical and hematological parameter values. There were differences seen in the basal values of creatine kinase (CK), urea, creatinine, aspartate transaminase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), potassium, total bilirubin, and eosinophil percentage compared to reference population data. However, other analytes showed narrow ranges of variation like glucose, total protein, albumin, sodium, hemoglobin, mean cell volume (MCV), and platelet count. Exercise produces changes in biochemical and hematological basal values of athletes compared to the general population, with the greatest variation in CK, but AST, ALT, LDH, potassium, and total bilirubin (TBil) show high values in serum, only with a wider distribution of values. The data here reflects the effect of exercise on biochemical and hematological parameter baseline ranges in elite athletes. As clinical laboratories use reference intervals to validate clinical reports, these "pseudo" reference intervals should be used when validating laboratory reports.


De Novo Maltotriose Biosynthesis from the Reducing End by Spinacia oleracea L. Chloroplasts.
The distribution of (14)C in the various glucose residues of maltotriose was studied as a function of time of photosynthesis of isolated chloroplasts of spinach (Spinacia oleracea L.) using (14)CO(2). The distribution of label showed that the reducing-end glucose residue was labeled first and the label subsequently distributed to the second and third glucose residues at approximately equal rates.A mechanism for the distribution of label and the synthesis of maltotriose from the reducing end is presented. The mechanism has postulated to be the same as that for the maltose synthase recently described by Schilling. Maltose biosynthesis from alpha-d-glucose-1-phosphate was characterized as involving two glucosyl intermediates by a double displacement mechanism with inversion of configuration. The mode of enzymic action by which maltosyl intermediates were transferred to glucosyl intermediates was consistent with the fractional distribution of radioactivity found in each glucose unit of maltotriose during short term photosynthesis experiments.


Monoclonal antibodies against protease inhibitors: the case of the bovine pancreatic trypsin inhibitor.
A monoclonal antibody (MoAb) directed against bovine basic pancreatic trypsin inhibitor (BPTI or aprotinin), a small protein made up by 58 aminoacids, has been produced. Mice were immunized with the native form of the protein and gave low antibody response. After somatic hybridization of spleen cells from immunized mice, few clones secreting antibodies against BPTI have been found and just two of them were stable. Both secreted IgM. One (ICI) produces a monoclonal antibody which binds to BPTI with an equilibrium dissociation constant, Kd, of 6.1 x 10(-7) M at pH 7.4. Competition experiments demonstrated that ICI recognizes an epitope close to the reactive site of BPTI. Furthermore, Kd of ICI with an isoinhibitor similar to BPTI (S.I. II) but with few differences at the active site is higher, confirming specificity of binding.


Polygenic dissection of the bipolar phenotype.
Recent data provide strong support for a substantial common polygenic contribution (i.e. many alleles each of small effect) to genetic susceptibility for schizophrenia and overlapping susceptibility for bipolar disorder. To test hypotheses about the relationship between schizophrenia and psychotic types of bipolar disorder. Using a polygenic score analysis to test whether schizophrenia polygenic risk alleles, en masse, significantly discriminate between individuals with bipolar disorder with and without psychotic features. The primary sample included 1829 participants with bipolar disorder and the replication sample comprised 506 people with bipolar disorder. The subset of participants with Research Diagnostic Criteria schizoaffective bipolar disorder (n = 277) were significantly discriminated from the remaining participants with bipolar disorder (n = 1552) in both the primary (P = 0.00059) and the replication data-sets (P = 0.0070). In contrast, those with psychotic bipolar disorder as a whole were not significantly different from those with non-psychotic bipolar disorder in either data-set. Genetic susceptibility influences at least two major domains of psychopathological variation in the schizophrenia-bipolar disorder clinical spectrum: one that relates to expression of a 'bipolar disorder-like' phenotype and one that is associated with expression of 'schizophrenia-like' psychotic symptoms.


Phosphorylation of c-Fos at the C-terminus enhances its transforming activity.
c-Fos is phosphorylated by MAP kinase and the 90 kDa-ribosomal S6 kinase (RSK) in vitro at serines 362 and 374 (rat) which we demonstrate are major in vivo phosphorylation sites in early G1. We have constructed c-Fos mutants with these serines changed to aspartic acid residues (FosD) to mimic phosphorylation or to alanine residues (FosA) to prevent phosphorylation. Cells expressing FosD exhibited a more extensive transformed phenotype than those expressing either FosA or wild type c-Fos (FosWT). We also observed that FosA has a reduced half-life in comparison with FosD in G1. Furthermore, we observed enhanced AP-1 transactivation activity in cells expressing FosD. These results indicate that phosphorylation of c-Fos at its extreme carboxyterminus, possibly by MAP kinase and RSK, supports the proliferative response by increasing c-Fos stability and/or by increasing its transactivation activity. Under conditions in which the MAP kinase pathway is constitutively activated, c-Fos phosphorylation probably contributes to cellular transformation. The highly conserved nature of these phosphorylation sites in other c-fos family members suggests that these may also be targets of MAP kinase and RSK.


Results Of Immediate Facial Nerve Reconstruction In Patients Undergoing Parotid Tumour Resection.
Facial nerve is usually sacrificed in total parotidectomy. The objective of this study is to present results of immediate reconstruction of facial nerve in total parotidectomy cases where facial nerve is sacrificed. This is a prospective study done in patients who had total parotidectomy including facial nerve and immediate reconstruction was done with inter-positional nerve grafts (sural n=12 and greater auricular n=10) from December 2017 till February 2018 by single surgeon (MR). Wounds were closed primarily (n=15), local flap (n=2) and free flap (n=5). Clinical evaluation was done at four months minimum follow up (those operated in January to February 2018) and eight months maximum follow up (those operated in December 2017), for facial nerve functional recovery using House and Brackmann grading system by single author (MR). Total of 22 (male n=7, female n=15) patients included in study from December 2017 till February 2018. Sural nerve grafts were used in 54% (n=12) and greater auricular nerve grafts in 45% (n=10) patients for reconstruction of facial nerve. On clinical evaluation using House and Brackmann grading system, showed grade V (n=4), grade IV (n=7), grade III (n=8) and grade II (n=3) repairs. Although primary end to end facial nerve repair is ideal but in situation where a significant segment of nerve is lost or where the repair is under tension, inter-positional nerve grafting is a simple and reliable reconstructive technique with good outcomes.


De novo generation of optically active small organic molecules using Monte Carlo tree search combined with recurrent neural network.
Optically active small organic molecules are computationally designed using the ChemTS python library developed by Tsuda and collaborators, which utilizes a combined Monte Carlo tree search (MCTS) and recurrent neural network model. Geometry optimization and excited-state calculations are performed for each generated molecule, following which the excitation energy and dissymmetry factors are computed to evaluate the score function in the MCTS process. Using this procedure, molecules not contained in existing databases are generated. Molecules having either high dissymmetry factors or high transition dipole strengths can be generated depending on the choice of the score function. In a single trajectory with 100,000 trials, mutually similar high-scoring molecules are generated frequently after the initial 15,000-20,000 trials. This indicates that it is better to sample high-scoring molecules from several trajectories having a modest number of trials each than from a single trajectory having a large number of trials.


Effect of small amounts of hydrogen added to argon glow discharges: hybrid Monte Carlo-fluid model.
A hybrid Monte Carlo-fluid modeling network is developed for an argon-hydrogen mixture, to predict the effect of small amounts of hydrogen added to a dc argon glow discharge. The species considered in the model include the Ar gas atoms, electrons, Ar(+) ions and fast Ar atoms, ArH(+), H(+), H(2)(+) and H(3)(+) ions, and H atoms and H(2) molecules, as well as Ar metastable atoms, sputtered Cu atoms, and the corresponding Cu(+) ions. Sixty-five reactions between these species are incorporated in the model. The effect of hydrogen on various calculation results is investigated, such as the species densities, the relative role of different production and loss processes for the various species, the cathode sputtering rate and contributions by different bombarding species, and the dissociation degree of H(2) and the ionization degree of Ar and Cu. The calculation results are presented and discussed for 1% H(2) addition, and comparison is also made with a pure argon discharge and with only 0.1% H(2) addition.


Overexpression of heparan sulfate 6-O-sulfotransferases in human embryonic kidney 293 cells results in increased N-acetylglucosaminyl 6-O-sulfation.
Heparan sulfate (HS) interacts with a variety of proteins and thus mediates numerous complex biological processes. These interactions critically depend on the patterns of O-sulfate groups within the HS chains that determine binding sites for proteins. In particular the distribution of 6-O-sulfated glucosamine residues influences binding and activity of HS-dependent signaling molecules. The protein binding domains of HS show large structural variability, potentially because of differential expression patterns of HS biosynthetic enzymes along with differences in substrate specificity. To investigate whether different isoforms of HS glucosaminyl 6-O-sulfotransferase (6-OST) give rise to differently sulfated domains, we have introduced mouse 6-OST1, 6-OST2, and 6-OST3 in human embryonic kidney 293 cells and compared the effects of overexpression on HS structure. High expression of any one of the 6-OST enzymes resulted in appreciably increased 6-O-sulfation of N-sulfated as well as N-acetylated glucosamine units. The increased 6-O-sulfation was accompanied by a decrease in nonsulfated as well as in iduronic acid 2-O-sulfated disaccharide structures. Furthermore, overexpression led to an altered HS domain structure, the most striking effect was the formation of extended 6-O-sulfated predominantly N-acetylated HS domains. Although the effect was most noticeable in 6-OST3-expressing cells, these results were largely independent of the particular 6-OST isoform expressed and mainly influenced by the level of overexpression.


Monitoring multiple distances within a single molecule using switchable FRET.
The analysis of structure and dynamics of biomolecules is important for understanding their function. Toward this aim, we introduce a method called 'switchable FRET', which combines single-molecule fluorescence resonance energy transfer (FRET) with reversible photoswitching of fluorophores. Typically, single-molecule FRET is measured within a single donor-acceptor pair and reports on only one distance. Although multipair FRET approaches that monitor multiple distances have been developed, they are technically challenging and difficult to extend, mainly because of their reliance on spectrally distinct acceptors. In contrast, switchable FRET sequentially probes FRET between a single donor and spectrally identical photoswitchable acceptors, dramatically reducing the experimental and analytical complexity and enabling direct monitoring of multiple distances. Our experiments on DNA molecules, a protein-DNA complex and dynamic Holliday junctions demonstrate the potential of switchable FRET for studying dynamic, multicomponent biomolecules.