Enzyme-responsive nano-drug delivery system for combined antitumor therapy.
Efficient drug loading, tumor targeting, intratumoral penetration, and cellular uptake are the main factors affecting the effectiveness of drug delivery systems in oncotherapy. Based on the tumor microenvironment, we proposed to develop Curcumin (Cur)-loaded matrix metalloproteinase (MMP)-responsive nanoparticles (Cur-P-NPs) by static electricity, to enhance tumor targeting, cellular uptake, and drug loading efficiency. These nanoparticles combine the properties of both PEG-peptides (cleaved peptide + penetrating peptide) and star-shaped polyester (DPE-PCL) nanoparticles. Cur-P-NPs displayed good entrapment efficiency, drug loading and biocompatibility. Additionally, they showed an enhanced release rate, cellular uptake, and anti-proliferative activity by activating peptides under the simulated tumor microenvironment. Furthermore, intraperitoneal injection of losartan (LST) successfully enhanced intratumoral drug penetration by collagen I degradation. In vivo studies based on the systematic administration of the synergistic LST + Cur-P-NPs combination to mice confirmed that combined antitumor therapy with LST and Cur-P-NPs could further improve intratumor distribution, enhance anticancer efficacy, and reduce the toxicity and side effects. Therefore, LST + Cur-P-NPs represent a new and efficient system for clinical oncotherapy.


SPECT/CT unequivocally depicts dilated cystic duct sign on hepatobiliary scintigraphy in acute cholecystitis.
The dilated cystic duct sign (DCDS) on hepatobiliary scintigraphy can lead to a false-negative test in patients with acute cholecystitis. Presented is a case of acute gangrenous cholecystitis with probable DCDS on the planar study that was unequivocally delineated on SPECT/CT imaging. SPECT/CT may be useful when planar findings suggest DCDS.


Morphology of the tongue dorsal surface of turbot (Scophthalmus maximus).
This study aimed to analyze the morphology of the tongue, which varies among fish species and has not been studied in turbot (Scophthalmus maximus), using macro-anatomical, light and scanning electron microscopy (SEM). As research materials, the tongues of eight adult turbot, which were used for consumption, were examined. The roughly triangular-shaped tongue of the turbot consisted of three parts: apex, body, and root. The thickness of the tongue continued to increase from the apex to the root. Although papillae were not observed in the macroscopic examination of the tongue, microscopic examination revealed the presence of cone-like shaped papillae on the submucosa. The tongue was composed of mucosa, submucosa, and hyaline cartilage. By SEM the presence of the taste buds (Types I and III), taste pores and papillae on the dorsal surface were demonstrated. Taste buds are the first descriptions in S. maximus. Therefore our results could add new data to the fish tongue anatomy. HIGHLIGHTS: Morphological and morphometric data of the turbot (Scophthalmus maximus) tongue were obtained in this study and compared with other species. In addition, the dorsal surface of the turbot tongue was described for the first time using SEM.


Magnesium homeostasis in Mammalian cells.
Magnesium, the second most abundant cation within the cell, plays an important role in numerous biological functions. Experimental evidence indicates that mammalian cells tightly regulate cellular magnesium ion content through specific mechanisms controlling Mg(2+) entry and efflux across the cell membrane and the membrane of various cellular organelles as well as intracellular Mg(2+) buffering under resting conditions and following hormonal and metabolic stimuli. This chapter will provide an assessment of the various mechanisms controlling cellular Mg(2+) homeostasis and transport, and the implications changes in cellular Mg(2+) content play under physiological and pathological conditions.


Protein phosphatase 1 activation and alternative splicing of Bcl-X and Mcl-1 by EGCG + ibuprofen.
Epigallocatechin-3-gallate (EGCG) and ibuprofen synergistically act to suppress proliferation and enhance apoptosis of prostate cancer cell lines, PC-3 and LNCaP. The purpose of this study was to investigate the mechanism of underlying this synergism. Most interestingly, EGCG + ibuprofen treatment in PC-3 cells resulted in altering the ratio of the splice variants of Bcl-X and Mcl-1, downregulating the mRNA levels of anti-apoptotic Bcl-X(L) and Mcl-1(L) with a concomitant increase in the mRNA levels of pro-apoptotic Bcl-X(s) and Mcl-1(s). However, there were no apparent changes in splicing variants in either ibuprofen or EGCG treated cells. Induction of alternative splicing was correlated with increased activity of protein phosphatase 1 (PP1) in EGCG + ibuprofen-treated cells, since pretreatment with calyculin A and tautomycin blocked EGCG + ibuprofen-induced alternative splicing in PC-3 cells in contrast to pretreatment with okadaic acid. On the other hand, EGCG + ibuprofen treatment in LNCaP cells did not alter splicing variants of Bcl-X and Mcl-1, despite the increase in protein phosphatase activity. In both cell lines, EGCG + ibuprofen inhibited cell proliferation synergistically. Taken together, this study demonstrate for the first time that EGCG + ibuprofen upregulated PP1 activity, which in turn induced alternative splicing of Bcl-X and Mcl-1 in a cell-type specific manner. Our study also demonstrates that the activation of PP1 contributes to the alternative splicing of Mcl-1.


Using visual processing training to enhance standard cognitive remediation outcomes in schizophrenia: A pilot study.
Approaches to cognitive remediation (CR) that address sensory perceptual skills before higher cognitive skills, have been found to be effective in enhancing cognitive performance in schizophrenia. To date, however, most of the conducted trials have concentrated on auditory processing. The aim of this study was to explore whether the addition of visual processing training could enhance standard cognitive remediation outcomes in a schizophrenia population. Twenty participants were randomised to either receive 20h of computer-assisted cognitive remediation alone or 20h of visual processing training modules and cognitive remediation training. All participants were assessed at baseline and at the end of cognitive remediation training on cognitive and psychosocial (i.e. self-esteem, quality of life) measures. At the end of the study participants across both groups improved significantly in overall cognition and psychosocial functioning. No significant differences were observed between groups on any of the measures. Of potential interest, however, was that the Cohen's d assessing the between group difference in the rates of change were moderate/large for a greater improvement in Visual Learning, Working Memory and Social Cognition for the visual training plus cognitive remediation group. On the basis of our effect sizes on three domains of cognition, we recommend replicating this intervention with a larger sample.


Efficacy of vaginal danazol treatment in women with menorrhagia during fertile age.
To evaluate the clinical efficacy and patient satisfaction of danazol delivered vaginally as treatment for young women with menorrhagia. Prospective study. University of Siena, Siena, Italy. Women with menorrhagia (n = 55; age range 25-35 years) after ultrasound and hysteroscopy. Low-dose danazol (200 mg/day) was daily administered by vaginal route for 6 months. Before and every month during the treatment women were requested: 1) to keep a diary of menstrual bleeding and to rate blood loss on a visual analog scale from 0 (no blood loss) to 10 (gushing-type bleeding); 2) to record side effects and their satisfaction with the therapy. Transvaginal ultrasound, blood count, serum chemistries, and serum concentration of LH, FSH, E(2), TSH, FT(3), FT(4), and PRL were evaluated before and after 6 months. The severity of blood loss was significantly reduced in all of the women after 2 months of treatment. Uterine volume was significantly reduced, and hematocrit, hemoglobin, and red blood cell count increased in all of the women after 6 months. The medical treatment did not affect hormonal parameters, and menstrual cycle remained unaffected; few local vaginal adverse effects were recorded. Vaginal danazol resulted in effective medical treatment in young women with menorrhagia, and, because of a lack of significant adverse effects, it may be proposed as an alternative treatment.


Regulation of macrophage number and gene transcript levels by activin A and its binding protein, follistatin, in the testes of adult mice.
The cytokine activin A is expressed throughout testicular development and is a critical regulator of macrophage function, but its effects on the testicular macrophages are not well-defined. Macrophage distribution and gene transcript levels were examined in testes of adult mice with reduced levels of either activin A (Inhba+/-), or its binding protein, follistatin (TghFST315). Macrophages were identified using F4/80 immunohistochemistry and enumerated by morphometry. Transcript levels were measured in testis extracts by qRT-PCR and Fluidigm ™ analyses. Interstitial macrophages were twice as numerous as peritubular macrophages in Inhba+/- and TghFST315 mice and their littermate controls. Macrophage numbers were significantly reduced in all regions of the Inhba+/- testis, and the volume density of peritubular and subcapsular macrophages was significantly reduced compared to littermate controls (by 52.9% and 36.3% respectively). Transcripts encoding macrophage chemokines, Csf1 and Ccl2, and receptor Csf1r, were elevated (by 35%, 44% and 27% respectively) in Inhba+/- testes, but Cx3cl1 and their receptors, Cx3cr1 and Ccr2, were not altered. Transcripts encoding MHC class II antigens and the co-stimulatory molecule Cd86, also increased (by 32% and 60% respectively), but other co-stimulatory molecules Cd80 and Cd274, and the scavenger receptor Mrc1 (CD206), were unaffected. In the follistatin-deficient testes, macrophage numbers and most macrophage-specific transcripts were not significantly affected, but Mrc1 expression was reduced by 35%. These data indicate that activin A maintains macrophage numbers, but selectively inhibits the levels of key transcripts associated with macrophage antigen-presentation, recruitment and differentiation in the adult mouse testis.


The value of the stentless biovalve prosthesis. An experimental study.
Clinical use of the stentless bioprosthesis has not yet been accepted world-wide. Experimental studies are scarce. In a study in growing pigs, 23 aortic valve replacements were performed (7 stented bioprostheses (STB), 10 stentless bioprostheses (SLB) and 6 cryopreserved homografts (CAH)). Valves were studied macroscopically and microscopically after explantation. Five animals died between 1.5 and 4 months after implantation. Two STBs showed extreme calcific degeneration. A third STB animal died suddenly: this STB showed only minimal leaflet calcification. One SLB was stenosed with a large fibrin deposit in one cusp, a second SLB showed slight cusp calcification and three were paracommissural tears in one cusp. In all other animals the valves were explanted after 5 to 6.5 months. All STBs showed severe calcific degeneration. Five SLBs showed mild calcific degeneration, while three others were unaltered after 6 months. All CAHs were free of calcific degeneration, three were perfect, two had a tear in a commisure and another was prolapsed with a fibrin degeneration. The speed and extent of valve degeneration were less than in the STB group. The stentless design is an important contribution which may result in a higher durability of bioprosthetic valves.


The HOXA1 A218G polymorphism and autism: lack of association in white and black patients from the South Carolina Autism Project.
A recent study has suggested that the A218G polymorphism in the homeobox A1 (HOXA1) gene may influence susceptibility to autism. We have determined the frequencies of the A and G alleles of the HOXA1 A218G polymorphism in both white and black patients from the South Carolina Autism Project (SCAP) and controls. Marked differences were found in allele frequencies between the races, but no deviations from Hardy-Weinberg equilibrium were seen in either white or black SCAP family members. More direct tests, comparing genotype frequencies between probands and controls and tracking transmission of the A versus G alleles to affected offspring, did not support the contention that allele status for the HOXA1 A218G polymorphism influences one's susceptibility to autism.


Frailty and Sarcopenia Assessment upon HospitalAdmission to Internal Medicine Predicts Length ofHospital Stay and Re-Admission: A ProspectiveStudy of 980 Patients.
Frailty and sarcopenia are associated with frequent hospitalizations and poor clinical outcomes in geriatric patients. Ascertaining this association for younger patients hospitalized in internal medicine departments could help better prognosticate patients in the realm of internal medicine. During a 1-year prospective study in an internal medicine department, we evaluated patients upon admission for sarcopenia and frailty. We used the FRAIL questionnaire, blood alanine-amino transferase (ALT) activity, and mid-arm muscle circumference (MAMC) measurements. We recruited 980 consecutive patients upon hospital admission (median age 72 years (IQR 65-79); 56.8% males). According to the FRAIL questionnaire, 106 (10.8%) patients were robust, 368 (37.5%) pre-frail, and 506 (51.7%) were frail. The median ALT value was 19IU/L (IQR 14-28). The median MAMC value was 27.8 (IQR 25.7-30.2). Patients with low ALT activity level (<17IU/L) were frailer according to their FRAIL score (3 (IQR 2-4) vs. 2 (IQR 1-3); p < 0.001). Higher MAMC values were associated with higher ALT activity, both representing robustness. The rate of 30 days readmission in the whole cohort was 17.4%. Frail patients, according to the FRAIL score (FS), had a higher risk for 30 days readmission (for FS > 2, HR = 1.99; 95CI = 1.29-3.08; p = 0.002). Frail patients, according to low ALT activity, also had a significantly higher risk for 30 days readmission (HR = 2.22; 95CI = 1.26-3.91; p = 0.006). After excluding patients whose length of stay (LOS) was ≥10 days, 252 (27.5%) stayed in-hospital for 4 days or longer. Frail patients according to FS had a higher risk for LOS ≥4 days (for FS > 2, HR = 1.87; 95CI = 1.39-2.52; p < 0.001). Frail patients, according to low ALT activity, were also at higher risk for LOS ≥4 days (HR = 1.87; 95CI = 1.39-2.52; p < 0.001). MAMC values were not correlated with patients' LOS or risk for re-admission. Frailty and sarcopenia upon admission to internal medicine departments are associated with longer hospitalization and increased risk for re-admission.


LC-MS/MS-based metabolite quantitation of the antiviral prodrug baloxavir marboxil, a new therapy for acute uncomplicated influenza, in human plasma: Application to a human pharmacokinetic study.
Baloxavir marboxil (BXM) is a novel orally administrated prodrug for the treatment of acute uncomplicated influenza. In the present study, a bioanalytical LC-MS/MS method was developed and validated for the quantification of baloxavir acid (BXA), the active form of baloxavir marboxil in plasma of healthy volunteers using dolutegravir as an internal standard (IS) following plasma protein precipitation with acetonitrile. BXA and the internal standard were chromatographically separated using Waters Xterra® MS C8 column (5 µm, 4.6 × 50 mm) and a mobile phase comprised of 10.0 mM ammonium formate pH 3.5 and acetonitrile (80:20, v/v) delivered at a flow rate of 0.6 mL/min. The transitions of m/z 484.00 → 247.0 and 420.30 → 277.1 for BXA and IS, respectively in multiple reaction monitoring (MRM) mode in a positive ESI interface were used for quantitation through triple-quad mass spectrometry, API 4000. The method linearity was proven across the concentration range of 0.5-200.0 ng/mL, adjusted, and validated completely in accordance with the bioanalytical guidelines of the United States-FDA. Finally, the present method was effectively applied for the pharmacokinetic study of BXA in healthy human volunteers with accepted reproducibility and ruggedness.


A three-dimensional reconstructive study of pelvic cavity in the red fox (Vulpes vulpes).
Three-dimensional (3D) reconstructive techniques are being used in many morphometric studies and clinical applications. The red fox is a wild animal localized in a wide geography. This study has been performed to determine morphometric properties of the pelvic cavity and reveal differences between sexes via 3D reconstruction obtained by multi-detector computed tomography images (MDCT) of the red fox. A total of 12 adult red foxes of both sexes were used in this study. Two-dimensional (2D) MDCT images of the pelvic cavity obtained and stored in DICOM format were used to reconstruct the 3D model. Mimics 13.1 program was used to perform the 3D reconstruction. A total of 24 dimensions were measured from the reconstructive model of the pelvic cavity. Twenty two dimensions were found to be significantly different (p < 0.05) between sexes. In conclusion, according to the pelvimetric data, it was determined that the red fox has sexual dimorphism and these data may be useful in wild animals morphological and gynaecological research. The authors hope to help in the diagnosis and treatment of the pelvic cavity-related orthopaedic and gynaecological disorders in the wild animals.


Roger Nicoll Tackles Learning/Disabilities.
Roger Nicoll was doubly honored by the Society for Neuroscience this year. He was awarded the Ralph W.


[Risk factors for developing pneumothorax in full-term neonates with respiratory distress].
To describe respiratory distress (RD) in full-term neonates hospitalized in the NICU and to determine risk factors in this population for pneumothorax. Retrospective inclusion for 4 years of full-term neonates hospitalized for RD before the 2nd day of life. Neonates were separated into Group I (RD with no pneumothorax) and Group II (RD with pneumothorax). Data collected from maternal and newborn medical records were obstetrical, perinatal, and postnatal. P<0.05 was set as the significance level. Ninety-six neonates were included. In this population, 64 (66.7%) were male, 45 (46.9%) were born by cesarean section, and 30 (31.3%) by elective cesarean section. Neonatal outcome was 4.6 days of hospital stay, 47.4% odds of mechanical ventilation, and 17.7% of persistent pulmonary hypertension of the neonate (PPHN). A central catheter was needed in 25% of the patients and amine treatment in 3.1%. The number of neonates born by cesarean section was lower as term increased. Those born by cesarean section were more likely to develop PPHN (26.7 vs 9.8%; P=0.03), and those born without labor were more likely to require oxygen (83.3 vs 63.6%; P=0.05). When comparing Group I and Group II (32 neonates), absence of labor (RR 1.5) and birth outside of a level III maternity unit (RR 1.6) were risk factors for pneumothorax. These results were confirmed in multivariate analysis. In Group II, birth before 39 weeks was a risk factor for bilateral pneumothorax (P=0.01). The median length of hospitalization was significantly longer in Group II than in Group I (5.8 days vs 4 days, P=0.03). RD at term exposes the infant to high morbidity and pneumothorax, especially if born outside of a level III maternity unit and absence of labor.


Toxicity Index, patient-reported outcomes, and persistence of breast cancer chemotherapy-associated side effects in NRG Oncology/NSABP B-30.
Adjuvant chemotherapy improves breast cancer survival but is associated with bothersome short- and long-term toxicity. Factors associated with toxicity, especially subacute toxicity up to 2 years following chemotherapy, have not been fully elucidated. The NRG Oncology/NSABP B-30 clinical trial compared 3 different doxorubicin-, cyclophosphamide-, and docetaxel-based chemotherapy regimens given over 3-6 months. Patients with hormone receptor-positive breast cancer received subsequent adjuvant endocrine therapy. From baseline through 24 months, 2156 patients completed questionnaires serially. We used multivariable probabilistic index models to identify factors associated with acute (>0-12 months) and subacute (>12-24 months) difficulties with pain, cognition, vasomotor symptoms, and vaginal symptoms. For all symptom domains, presence of symptoms prior to chemotherapy initiation were associated with symptoms in the subacute period (all p < 0.001). In addition, different combinations of patient factors and breast cancer treatments were associated with increased likelihood of pain, vasomotor, and vaginal symptoms in the subacute period. Consideration of pre-treatment symptoms and patient factors, as well as treatments for breast cancer, can facilitate identification of groups of patients that may experience symptoms following completion of chemotherapy. This information may be important for treatment-decision-making when alternative regimens are equivalent in benefit.


Use of DWI-only MR protocol for screening stroke mimics.
Patients presenting with focal neurological symptoms may suffer from stroke or stroke mimics. Diffusion weighted MRI (DWI) is highly sensitive for identifying acute ischemia. Therefore, we aimed to explore whether a DWI-only protocol would help differentiate stroke from stroke mimics. We identified all patients with possible but not definite stroke that underwent DWI-only MRI between 6/2010 and 8/2011. Patients with a positive DWI lesion were compared to those with negative DWI findings on demographics, risk factor profile, final discharge diagnoses, and outcome. A total of 124 patients were included with a median age of 63.5 (53% male). DWI MRI was positive for acute ischemia in 46 patients (37%). The most frequent stroke mimics were peripheral vertigo (n=19), acute confusion (n=10), seizures (n=9) and migraine with aura (n=8). Most ischemic lesions were small on DWI (<2 cm) and patients had minor disability (mean NIHSS 4.9±3.9) with 81% of patients having good outcomes (modified Rankin Score≤2) at 3 months. On univariate analysis patients with positive DWI studies had higher frequencies of having more than one clinical symptom (56% vs. 13% respectively; P<0.001) and this variable remained a significant predictor for stroke on multivariate analysis (OR 9.4 95% CI 3.8-23.5). A short DWI-only MRI protocol can effectively differentiate stroke from stroke mimics and could be used in settings of the emergency department as well as later on for diagnostic purposes. The chances for finding positive DWI lesions are increased in patients with multiple symptoms and signs.


Most Short Children with Cystic Fibrosis Do Not Catch Up by Adulthood.
Poor linear growth is common in children with cystic fibrosis (CF) and predicts pulmonary status and mortality. Growth impairment develops in infancy, prior to pulmonary decline and despite aggressive nutritional measures. We hypothesized that growth restriction during early childhood in CF is associated with reduced adult height. We used the Cystic Fibrosis Foundation (CFF) patient registry to identify CF adults between 2011 and 2015 (ages 18-19 y, n = 3655) and had height for age (HFA) records between ages 2 and 4 y. We found that only 26% CF adults were ≥median HFA and 25% were <10th percentile. Between 2 and 4 years, those with height < 10th percentile had increased odds of being <10th percentile in adulthood compared to children ≥ 10th percentile (OR = 7.7). Of HFA measured between the 10th and 25th percentiles at ages 2-4, 58% were <25th percentile as adults. Only 13% between the 10th and 25th percentile HFA at age 2-4 years were >50th percentile as adults. Maximum height between ages 2 and 4 highly correlated with adult height. These results demonstrate that low early childhood CF height correlates with height in adulthood. Since linear growth correlates with lung growth, identifying both risk factors and interventions for growth failure (nutritional support, confounders of clinical care, and potential endocrine involvement) could lead to improved overall health.


Expression of the bovine aromatase cytochrome P450 gene (Cyp19) is primarily regulated by promoter 2 in bovine follicles and by promoter 1.1 in corpora lutea.
Follicular development and differentiation are closely associated with increasing steroidogenesis. During the present study transcript concentration of Cyp19, Cyp11A1, and 3beta-hydroxysteroid dehydrogenase delta (3beta-HSD) encoding the steroidogenic enzymes P450(arom), P450(SCC), and 3beta-HSD were determined by real-time PCR in bovine granulosa cells (GC) as potential markers for follicular differentiation. Ovaries were collected from a local abattoir (experiment 1) and from synchronized animals at day 4 of estrus cycle (experiment 2). To study effects of luteinization, steroidogenic transcripts were also quantified in corpora lutea (CL) 4 and 20 days after fertilization. In most follicles, all three steroidogenic transcripts were detected, however, at very different concentration. Expression of 3beta-HSD and Cyp11A1 was highly significantly co-regulated and showed a significant correlation with follicular size. Contrary, Cyp19 expression was extremely variable even in follicles of similar size. Cyp19 transcripts were derived predominantly from promoter P2 and less abundant from promoters P1.1 and P1.5. After luteinization, the concentration of 3beta-HSD and Cyp11A1 transcripts increased (75-fold and fivefold, respectively) whereas the Cyp19 transcript level dropped (160-fold). Residual Cyp19 transcripts in CL were almost exclusively derived from P1.1. The data indicate that Cyp19 expression in GC is predominantly regulated by P2 and to a minor extend by P1.1, whereas P1.1 is almost exclusively responsible for residual Cyp19 expression in CL. Correlation analyses suggest that the expression of 3beta-HSD and Cyp11A1 primarily depend on the size of follicles whereas the concentration of P2 derived Cyp19 transcripts in GC is a marker for follicular differentiation towards selection and dominance.


[Pharmacokinetics of tacrolimus following topical application of tacrolimus ointment in adult and paediatric patients with atopic dermatitis].
Safety concerns related to systemic immunosuppressive therapy have raised questions regarding systemic exposure to topically applied tacrolimus. To summarise all currently available information on the pharmacokinetics of tacrolimus ointment and to compare it with the pharmacokinetics of systemic use of tacrolimus. Low and highly variable systemic exposure has been shown in pharmacokinetic studies of tacrolimus ointment performed in patients (adults and children) with moderate to severe atopic dermatitis. Patients with larger treatment areas tended to have higher exposure to the drug, but there was no evidence of systemic accumulation. Overall, 96, 92 and 97% of the blood samples assayed contained tacrolimus concentrations below 1 ng/ml, and 23, 17 and 20% of samples were below 0.025 ng/ml (the lower limit of quantification) in adults, children aged 6-12 and infants aged 3-24 months, respectively. Although tacrolimus applied topically is absorbed systemically, the overall exposure as measured by the area under the curve (AUC)0-24 is low and highly variable. The extent of systemic exposure is higher in patients with larger treatment areas, but even in patients with up to 75% of the body surface area treated, the systemic exposure is substantially lower than that measured in transplant patients. Systemic exposure to tacrolimus tends to decrease as the skin lesions heal and there is no evidence of systemic accumulation despite repeated applications of ointment. The tacrolimus ointment was efficacious and well-tolerated.


[External respiration function in patients with exudative pleurisy before and after video-thoracoscopic operations].
57 patients with exudative pleurisy were observed. Function of external respiration was examined with traditional spirometry and a new method--distant spirometry. Function of cardio-vascular system was examined with electrocardiography and echocardiography. The new method of monitoring permits to perform repeated and daily examinations of external respiration function. The method is highly informative, sabe, physiological, permits to evaluate general and isolated ventilation of the lungs simultaneously, and demonstrates dynamics of exudate accumulation that may help in choice of treatment policy.


Hepatitis C viral load, genotype, and increased risk of developing end-stage renal disease: REVEAL-HCV study.
The association between hepatitis C virus (HCV) infection and end-stage renal disease (ESRD) remains controversial without considering the role of HCV viral load and genotype. This study aimed to determine whether HCV RNA level and genotype affect the risk of developing ESRD. Between 1991 and 1992, 19,984 participants aged 30-65 years were enrolled in a community-based prospective cohort study in Taiwan. Chronic HCV infection was defined by detectable HCV viral load. ESRD was determined as the need for chronic dialysis or renal transplantation. Conventional Cox proportional hazard and competing risk models were used to determine the hazard ratio (HR) for ESRD. After a median follow-up of 16.8 years, 204 cases were detected during 319,474 person-years. The incidence rates of ESRD for nonchronically HCV-infected and chronically HCV-infected patients were 60.2 and 194.3 per 100,000 person-years, respectively. The multivariable HR was 2.33 (95% confidence interval [CI] 1.40-3.89) when comparing patients with and without chronic HCV infection. Patients with low and high HCV RNA levels were at higher risk of ESRD than those who were nonchronically HCV-infected (HR, 2.11, 95% CI 1.16-3.86, and HR, 3.06, 95% CI 1.23-7.58; Ptrend < 0.001). This association remained robust after taking pre-ESRD death as a competing event for ESRD. Patients with HCV genotype 1 tended to have a higher risk of developing ESRD (HR, 3.60 95% CI 1.83-7.07) compared with nonchronically HCV-infected subjects. This study reveals that chronic HCV infection is associated with an increased risk of developing ESRD and suggests that elevated serum levels of HCV RNA (>167,000 IU/mL) and HCV genotype 1 are strong predictors of ESRD, indicating clinical implications for the management of chronic HCV. (Hepatology 2017;66:784-793).


Characterization of soy-deprestatin, a novel orally active decapeptide that exerts antidepressant-like effects via gut-brain communication.
We found that the orally administered thermolysin digest of β-conglycinin exhibits antidepressant-like effects in tail suspension and forced swim tests in mice. A comprehensive peptide analysis of the digest using liquid chromatography/mass spectrometry was performed, and LSSTQAQQSY emerged as a candidate antidepressant-like peptide. Orally administered synthetic LSSTQAQQSY exhibited antidepressant-like effects at a dose of 0.3 mg/kg; therefore, we named the decapeptide soy-deprestatin. In contrast, intraperitoneally administered soy-deprestatin was ineffective. We then hypothesized that it acted on the gut, and its signal was transferred to the brain. Indeed, orally administered soy-deprestatin exhibited antidepressant-like activity in sham-treated, but not vagotomized, mice. Oral administration of soy-deprestatin also increased the c-Fos expression in the nucleus of the solitary tract, which receives inputs from the vagus nerve. These results suggested that the antidepressant-like effects were mediated by the vagus nerve. Thermolysin digest- and soy-deprestatin-induced antidepressant-like effects were also blocked by antagonists of serotonin 5-HT1A, dopamine D1, or GABAA receptors. We also clarified the order of receptor activation as 5-HT1A, D1, and GABAA, using selective agonists and antagonists. Taken together, soy-deprestatin may exhibit antidepressant-like effects after oral administration via a novel pathway mediated by 5-HT1A, followed by D1 and GABAA systems. This is the first orally active peptide demonstrating antidepressant-like effects via gut-brain communication.-Mori, Y., Asakura, S., Yamamoto, A., Odagiri, S., Yamada, D., Sekiguchi, M., Wada, K., Sato, M., Kurabayashi, A., Suzuki, H., Kanamoto, R., Ohinata, K. Characterization of soy-deprestatin, a novel orally active decapeptide that exerts antidepressant-like effects via gut-brain communication.


Hereditary fructose intolerance in childhood. Diagnosis, management, and course in 55 patients.
The early manifestations of hereditary fructose intolerance are described in a series of 55 patients. Management of this metabolic disorder depends on the severity of liver impairment. When the patients are given a fructose-free diet, the improvement is a dramatic but liver enlargement and fatty vacuolization of liver cells often persist. These hepatic findings were also observed in the five homozygous infants who were given a fructose-free diet from birth; this outcome may support the hypothesis that minimal amounts of fructose are esential for human beings.


Functional E3 ligase hotspots and resistance mechanisms to small-molecule degraders.
Targeted protein degradation is a novel pharmacology established by drugs that recruit target proteins to E3 ubiquitin ligases. Based on the structure of the degrader and the target, different E3 interfaces are critically involved, thus forming defined 'functional hotspots'. Understanding disruptive mutations in functional hotspots informs on the architecture of the assembly, and highlights residues susceptible to acquire resistance phenotypes. Here we employ haploid genetics to show that hotspot mutations cluster in substrate receptors of hijacked ligases, where mutation type and frequency correlate with gene essentiality. Intersection with deep mutational scanning revealed hotspots that are conserved or specific for chemically distinct degraders and targets. Biophysical and structural validation suggests that hotspot mutations frequently converge on altered ternary complex assembly. Moreover, we validated hotspots mutated in patients that relapse from degrader treatment. In sum, we present a fast and widely accessible methodology to characterize small-molecule degraders and associated resistance mechanisms.


Does temperature favour the spread of Raphidiopsis raciborskii, an invasive bloom-forming cyanobacterium, by altering cellular trade-offs?
As a tropical filamentous cyanobacterium, Raphidiopsis raciborskii has attracted much attention due to its expansion and toxin production. However, the mechanisms of its expansion to temperate regions have not been studied in detail. To address the potential strategies, the physiological and metabolomic profiles of R. raciborskii FACHB 1096 isolated from a temperate lake in China were determined and measured at different temperatures (10 °C, 15 °C, 20 °C, 25 °C, and 32 °C). The results demonstrated that temperature significantly changed cell viability, chlorophyll a content, specific growth rate, Chl a fluorescence, and filamentous shape of R. raciborskii. Low temperature decreased cell viability, specific growth rate, and photosynthetic efficiency, while the proportion of akinete and carbon fixation per unit cell were significantly increased compared with high temperature (32 °C). A constructed unimodal model indicated that filament length, cell volume, and cell length/width of R. raciborskii were significantly reduced in both high and low temperature environments. Under low-temperature conditions, R. raciborskii suffered different degrees of oxidative damage and produced corresponding antioxidant substances to resist oxidative stress, suggesting that low temperature changes the metabolic level of the cells, causing the cells to gradually switch from development to defense. Metabolomic data further confirmed that temperature change induced shifts in metabolic pathways in R. raciborskii, including starch and sucrose metabolic pathways, glutathione metabolic pathways, and the pentose phosphate pathways (PPP), as well as metabolic pathways related to the tricarboxylic acid (TCA) cycle. Our results indicated that the trade-offs of R. raciborskii cells among the growth, cell size, and metabolites can be significantly regulated by temperature, with broad implications for its global expansion in temperate waterbodies.


Supraceliac aortomesenteric bypass for intestinal ischemia.
The supraceliac aorta has received little attention as an inflow site of bypass graft origin for mesenteric arterial reconstruction; this article describes our experience with its use in 10 patients. Ten patients underwent bypass grafts (three vein and seven prosthetic grafts) from the supraceliac aorta incorporating the celiac axis to the superior mesenteric artery. All patients had two- or three-vessel mesenteric arterial occlusion. No operative deaths occurred. Nine patients were studied with postoperative arteriography. Nine of ten patients have had satisfactory results on follow-up from 5 to 99 months (mean 43 months), although two patients required operative revision to achieve this. The operative technique is described, including approach to the aorta in the lower mediastinum via upper abdominal exposure and pancreatic displacement to expose the superior mesenteric artery. This method permits avoidance of the infrarenal aorta, if desired because of atherosclerotic disease or previous operation. Antegrade supraceliac aortomesenteric grafts avoid turbulence and compression of conventional "retrograde" bypass beneath the mesentery. We conclude that supraceliac aortic grafts provide a useful visceral artery reconstruction technique.


Snowmelt periods as hot moments for soil N dynamics: a case study in Maine, USA.
The vernal transition represents the seasonal transition to spring, occurring as temperatures rise at the end of winter. With rapid snowmelt, microbial community turnover, and accelerated nutrient cycling, this is a critical but relatively under-studied period of ecosystem function. We conducted a study over two consecutive winters (2015-2016) at the Bear Brook Watershed in Maine to examine how changing winter conditions (warming winters, reduced snow accumulation) altered soil nitrogen availability and stream N export during winter and the vernal transition, and how these patterns were influenced by ecosystem N status (N-enriched vs. N-limited). Of the two study years, 2016 had a warmer winter with substantially less snow accumulation and a discontinuous snowpack-and as a result, had a longer vernal transition and a snowpack that thawed before the vernal transition began. Across both years, snowmelt triggered a transition, signaled by increased ammonium concentrations in soil, decreased soil nitrate concentrations due to flushing by meltwater, and increased stream nitrate exports. Despite the contrasting winter conditions, both years showed similar patterns in N availability and export, differing only in the timing of these transitions. The vernal transition has conventionally been considered a critical period for biogeochemical cycling, because the associated snowmelt event triggers physicochemical and biochemical changes in soil systems. This was consistent with our results in 2015, but our data for 2016 show that this may not always hold true, and instead, that warmer, low-snow winters may demonstrate a temporal asynchrony between snowmelt and the vernal transition. We also show that ecosystem N status is a strong driver of the seasonal N pattern, and the interaction of N status and changing climate must be further investigated to understand ecosystem function under our current predicted trajectory of warming winters, declining snowfall, and winter thaw events.


A quality of life survey of individuals with urinary incontinence who visit a self-help website: implications for those seeking healthcare information.
Urinary Incontinence (UI) affects 200 million people worldwide with annual direct costs in the US alone estimated at $16.3 billion. Those with UI have reported a decrease in general quality of life with symptoms of depression, anxiety, low self-esteem, poor body image, and social stigmatization. The purpose of this study was to examine the feasibility of collecting self-reported quality of life data in a self-selected sample of individuals who visited a website providing information, education, and management suggestions regarding UI. Participants included 374 individuals with UI who responded to a solicitation for enrollment in a "Continence Comprehensive Health and Life Assessment" survey posted on The Simon Foundation for Continence website (www.simonfoundation.org). Types of problems and events associated with UI, including social connectivity and quality of life, are discussed along with limitations of the study and implications for future research. Given that 13.01% of respondents had not spoken to a healthcare provider about their UI symptoms, 24.73% had never seen a healthcare professional who "specializes in bladder problems," and 75% said they were not currently using any active approach to managing symptoms, use of such information is discussed in terms of how to construct internet healthcare information to maximize seeking appropriate healthcare services and preparing internet-based information regarding incontinence diagnosis and treatment.


Multispectral imaging detects gastritis consistently in mouse model and in humans.
Gastritis constitutes the initial step of the gastric carcinogenesis process. Gastritis diagnosis is based on histological examination of biopsies. Non-invasive real-time methods to detect mucosal inflammation are needed. Tissue optical properties modify reemitted light, i.e. the proportion of light that is emitted by a tissue after stimulation by a light flux. Analysis of light reemitted by gastric tissue could predict the inflammatory state. The aim of our study was to investigate a potential association between reemitted light and gastric tissue inflammation. We used two models and three multispectral analysis methods available on the marketplace. We used a mouse model of Helicobacter pylori infection and included patients undergoing gastric endoscopy. In mice, the reemitted light was measured using a spectrometer and a multispectral camera. We also exposed patient's gastric mucosa to specific wavelengths and analyzed reemitted light. In both mouse model and humans, modifications of reemitted light were observed around 560 nm, 600 nm and 640 nm, associated with the presence of gastritis lesions. These results pave the way for the development of improved endoscopes in order to detect real-time gastritis without the need of biopsies. This would allow a better prevention of gastric cancer alongside with cost efficient endoscopies.


A 65-Year-Old Groundskeeper With High Fever, Pulmonary Nodules, and Thoracic Lymphadenopathy.
A 65-year-old man with treated latent tuberculous infection presented with 1 week of fevers (up to 39.6°C), chills, headache, lightheadedness, and malaise. He reported a chronic, nonproductive cough without hemoptysis but denied other localizing symptoms, sick contacts, or recent travel. He lived in an urban area in eastern Colorado and owned one healthy dog but otherwise denied known animal exposures. He was a retired oil driller who had worked in southern Arizona, New Mexico, and northern Mexico (Sonora region). Other travel included 3 years in the early 1970s working as a military aircraft mechanic in Vietnam, Laos, and Thailand. Six weeks prior to admission, he began work as a groundskeeper on a golf course that had experienced recent flooding, using a riding mower and exposing himself to airborne dust and organic debris. He smoked a pipe daily for 30 years but quit 2 months prior to presentation, although he continued to smoke marijuana weekly. He denied intravenous drug use.


Medial Protrusio Technique Versus Structural Autologous Bone-Grafting Technique in Total Hip Arthroplasty for Crowe Type II to III Hip Dysplasia.
It is unclear whether acetabular reconstruction techniques have any impact on clinical outcomes. This study aimed to determine (1) whether acetabular reconstruction techniques influenced the position of the acetabular cup and (2) whether clinical outcomes based on the acetabular reconstruction techniques differ in patients undergoing total hip arthroplasty (THA) with Crowe II to III developmental dysplasia of the hip. This was a retrospective analysis of prospectively collected data from 69 patients (74 hips) who were treated with cementless THA using medial protrusio technique (MPT) or structural autologous bone-grafting technique (SABT). There were 39 patients (41 hips) included in the MPT group and 30 patients (33 hips) in the SABT group. Clinical and radiographic outcomes were evaluated. All patients were followed up for at least 3 years. There were similar results between the 2 groups in terms of blood loss, Harris hip score, leg length discrepancy, cup inclination, cup anteversion, and proportion of cup coverage (P > .05). The operative time was significantly longer in the SABT group compared with the MPT group (P < .001). The postoperative vertical center of rotation was significantly higher in the MPT group compared with the SABT group (P = .001), and postoperative horizontal center of rotation was significantly shallower in the SABT group compared with the MPT group (P < .001). The MPT and SABT provide similar clinical and radiographic outcomes in the management of Crowe II to III developmental dysplasia of the hip by cementless THA. However, the MPT has the advantage of a shorter operative time, whereas the SABT is more conducive to placing the acetabular cup in an anatomic position. Level III, Therapeutic, Case-Control Study.


Impact of intermittent portal clamping on the early recurrence of hepatocellular carcinoma after surgery.
To evaluate the impact of intermittent portal clamping (IPC) during surgery on the early recurrence of hepatocellular carcinoma (HCC). The subjects of this retrospective study were 266 patients who underwent curative liver resection for HCC. The patients were grouped as follows: an intermittent portal clamping (IPC) group, n = 78; a continuous portal clamping (CPC) group, n = 128; and a non-portal clamping (NPC) group, n = 60. The median recurrence-free interval within 2 years of follow-up was significantly shorter in the IPC group (14.2 ± 4.6 months) than in the CPC group (18.0 ± 4.8 months, P = 0.008) or the NPC group (19.04 ± 4.1 months, P = 0.023). Moreover, 2-year recurrence-free survival was much lower in the IPC group than in the CPC group (63.6 vs. 75.8 %, P = 0.025) or the NPC group (63.6 vs. 78.0 %, P = 0.030). However, the 2-year OS rate among the three groups was comparable (72.7 vs. 79.9 %; P = 0.101) and 83.1 %, (P = 0.125). According to univariable analysis, tumor size (>5 cm), tumor number (≥2), tumor grade (low/undifferentiated), TNM stage (III), vascular infiltration, blood transfusion, and IPC were significantly associated with the early postoperative recurrence of HCC. After multivariate analysis, significance of tumor grade (low/undifferentiated) and TNM stage (III) disappeared, whereas tumor size (>5 cm), tumor number (≥2), vascular infiltration, blood transfusion, and IPC remained significant. IPC is an independent risk factor for the early recurrence of HCC after surgery.


Pituitary adenylate cyclase activating peptide deficient mice exhibit impaired thymic and extrathymic regulatory T cell proliferation during EAE.
We have shown that mice deficient in pituitary adenylate cyclase-activating polypeptide (PACAP, gene name ADCYAP1) manifest enhanced sensitivity to experimental autoimmune encephalomyelitis (EAE), supporting the anti-inflammatory actions described for this neuropeptide. In addition to an increased proinflammatory cytokine response in these mice, a reduction in regulatory T cell (Treg) abundance in the lymph nodes (LN) was observed, suggesting altered Treg kinetics. In the present study, we compared in PACAP deficient (KO) vs. wild type mice the abundances and rates of proliferation FoxP3(+) Tregs in three sites, the LN, central nervous system (CNS) and thymus and the relative proportions of Th1, Th2, and Th17 effector subsets in the LN and CNS. Flow cytometry analyses revealed a decrease in Treg proliferation and an increased T effector/Tregs ratio in the LN and CNS of PACAP KO mice. In the thymus, the primary site of do novo natural Treg production, the total numbers and proliferative rates of FoxP3(+) Tregs were significantly reduced. Moreover, the expression of IL-7, a cytokine implicated in thymic Treg expansion during EAE, failed to increase at the peak of the disease in the thymus and LN of PACAP KO mice. In addition to these Treg alterations, a specific reduction of Th2 cells (about 4-fold) was observed in the lymph nodes in PACAP KO mice, with no effects on Th1 and Th17 subsets, whereas in the CNS, Th1 and Th17 cells were increased and Th2 decreased. Our results suggest that endogenous production of the neuropeptide PACAP protects against EAE by modulating Treg expansion and Th subsets at multiple sites.


[Agar tube method for the bioassay of antibiotics (author's transl)].
A simplified bioassay system for antibiotics has been presented. Glass tubes with open ends and pits are filled with agar medium containing test microbes by immersing vertically into the agar medium while it is warm. These agar tubes are inserted in test solutions and incubated. The growth-inhibitory zones appear in respective tubes due to upward diffusion of the antibiotic. The heights of such zones from the bottom of agar tubes are measured.


R loops: from transcription byproducts to threats to genome stability.
RNA:DNA hybrid structures known as R loops were thought to be rare byproducts of transcription. In the last decade, however, accumulating evidence has pointed to a new view in which R loops form more frequently, impacting transcription and threatening genome integrity as a source of chromosome fragility and a potential cause of disease. Not surprisingly, cells have evolved mechanisms to prevent cotranscriptional R loop formation. Here we discuss the factors and cellular processes that control R loop formation and the mechanisms by which R loops may influence gene expression and the integrity of the genome.


The Burden and Management of Dental Caries in Older Children.
Dental caries is endemic in children and adolescents and has significant morbidity. This complex and chronic disease has both genetic and environmental etiologic factors. In children the preponderance of caries affects tooth surfaces with pits and fissures despite these representing only a small portion of the tooth surfaces that are at risk. Pit and fissure sealants are effective in preventing and managing noncavitated caries lesions in these surfaces. A variety of materials are clinically effective, and health care guidelines recommend the use of pit and fissure sealants as part of a comprehensive dental caries prevention program.


Analysis of age-associated alternation of SCSA sperm DNA fragmentation index and semen characteristics of 1790 subfertile males in China.
It has been identified that incidence of infertility was about 20% among couples worldwide, about 50% caused by male elements. However, conventional semen laboratory detections could not handle clinical needs, which led to more comprehensive parameters for male fertility evaluation. We aimed to investigate the clinical relationship of age-linked changes and the sperm chromatin structure assay (SCSA) sperm DNA fragmentation index (DFI), and routine semen characteristics among subfertile Chinese males. 1790 clinical semen specimens were enrolled from February 2018 to October 2019. Clinical and laboratory data including routine semen analyses, sperm DFI, and sperm morphology were collected and showed age-related alterations in semen parameters. Our results, displayed an increase in sperm DFI with age, were demonstrated in three age-groups, particularly within the ≥35-year cohort. There were positive and inverse correlations of sperm DFI with abnormal semen characteristics and with normal morphological parameters, respectively. Furthermore, age, sperm morphology, concentration, and progressive motility, immotile sperm percentage, semen volume, sperm survival, and high acridine orange DNA stainability (indicating immature forms) were found to be independent risk factors affecting sperm DNA integrity. Likewise, men aged ≥35 years had a higher sperm DFI than did normozoospermic men in the overall cohort. Routine semen characteristics, sperm DFI, and morphology tended to alter with age. The SCSA sperm DFI showed the greatest clinical application in the assessment of male fertility in this study, which should help infertility clinics decide on reproductive options for the treatment of older infertile couples.


Lymphocyte function-associated antigens one (LFA-1) on B and on T lymphocytes bind a monoclonal antibody with different affinities.
The kinetics of the binding of an anti-LFA-1 monoclonal antibody to lymphocytes of the B and T lineages was studied. A Kd approximately ten times lower was observed when binding the antibody to B splenocytes compared to T splenocytes. Using Fab fragments and measuring the dissociation rate constants gave a similar difference in binding kinetics of anti-LFA-1 between the different splenocytes. This difference of cell-binding affinity was independent of the state of stimulation of the cells by lipopolysaccharide or concanavalin A. Thymocytes reacted with the antibody with the same Kd as T splenocytes. The combined results indicate that LFA-1 from B and from T lymphocytes have a different conformation. Binding with a higher affinity to B and a lower affinity to T lymphocytes could also be demonstrated using liposomes coated with anti-LFA-1 antibody.


Mechanisms that match ATP supply to demand in cardiac pacemaker cells during high ATP demand.
The spontaneous action potential (AP) firing rate of sinoatrial node cells (SANCs) involves high-throughput signaling via Ca(2+)-calmodulin activated adenylyl cyclases (AC), cAMP-mediated protein kinase A (PKA), and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII)-dependent phosphorylation of SR Ca(2+) cycling and surface membrane ion channel proteins. When the throughput of this signaling increases, e.g., in response to β-adrenergic receptor activation, the resultant increase in spontaneous AP firing rate increases the demand for ATP. We hypothesized that an increase of ATP production to match the increased ATP demand is achieved via a direct effect of increased mitochondrial Ca(2+) (Ca(2+)m) and an indirect effect via enhanced Ca(2+)-cAMP/PKA-CaMKII signaling to mitochondria. To increase ATP demand, single isolated rabbit SANCs were superfused by physiological saline at 35 ± 0.5°C with isoproterenol, or by phosphodiesterase or protein phosphatase inhibition. We measured cytosolic and mitochondrial Ca(2+) and flavoprotein fluorescence in single SANC, and we measured cAMP, ATP, and O₂ consumption in SANC suspensions. Although the increase in spontaneous AP firing rate was accompanied by an increase in O₂ consumption, the ATP level and flavoprotein fluorescence remained constant, indicating that ATP production had increased. Both Ca(2+)m and cAMP increased concurrently with the increase in AP firing rate. When Ca(2+)m was reduced by Ru360, the increase in spontaneous AP firing rate in response to isoproterenol was reduced by 25%. Thus, both an increase in Ca(2+)m and an increase in Ca(2+) activated cAMP-PKA-CaMKII signaling regulate the increase in ATP supply to meet ATP demand above the basal level.


Comparative glycomics of connective tissue glycosaminoglycans.
Homeostasis of connective joint tissues depends on the maintenance of an extracellular matrix, consisting of an integrated assembly of collagens, glycoproteins, proteoglycans, and glycosaminoglycans (GAGs). Isomeric chondroitin sulfate (CS) glycoforms differing in position and degree of sulfation and uronic acid epimerization play specific and distinct functional roles during development and disease onset. This work profiles the CS epitopes expressed by different joint tissues as a function of age and osteoarthritis. GAGs were extracted from joint tissues (cartilage, tendon, ligment, muscle, and synovium) and partially depolymerized using chondroitinase enzymes. The oligosaccharide products were differentially stable isotope labeled by reductive amination using 2-anthranilic acid-d(0) or -d(4) and subjected to amide-hydrophilic interaction chromatography (HILIC) online LC-MS/MS. The analysis presented herein enables simultaneous profiling of the expression of nonreducing end, linker region, and Delta-unsaturated interior oligosaccharide domains of the CS chains among the different joint tissues. The results provide important new information on the changes to the expression of CS GAG chains during disease and development.


Historical records of heavy metal accumulation in sediments and the relationship with agricultural intensification in the Yangtze-Huaihe region, China.
The extensive use of fertilizers on farmlands during the past several decades in China has led to a rapid deterioration of environmental water quality in recent years. An agricultural watershed with high-yield grain production and characterized by multipond systems in the Yangtze-Huaihe region of China was selected to establish the historic records of heavy metal pollution by (137)Cs-dated sediment cores. The experimental results indicated that the contents of most of the heavy metals investigated, such as Cd, Cr, Cu, Ni, Pb, and Zn, continuously increased in the multipond sediments throughout the past three decades. An inflection point appeared in the 1980s, prior to which all heavy metal contents showed little or no increase with time. Thereafter, the heavy metal contents increased dramatically due to the extensive application of phosphate fertilizers. The mean Cd content in 1980 was 0.13 microg/g which increased rapidly to 0.33 microg/g in 2004, but prior to 1980, the level was only 0.08 microg/g. Similar trends were also found for other heavy metals. The enrichment factor (EF) values indicated that Cd was not only the most anthropogenic metal in the multipond sediments but also aggravated anthropogenic impacts on the watershed environment during the past two decades. The high proportion of chemical reactive forms of Cd (10%-30%) implied a moderately high ecological risk. Our results clearly reveal that the extensive use of fertilizers has resulted in significant heavy metal pollution in this watershed, which threatens the water quality of the watershed and downstream water bodies.


Phthalates, physical activity, and diet, which are the most strongly associated with obesity? A case-control study of Chinese children.
Phthalate esters have been a research hotspot recently owing to potential obesogenic activity, but conflicting results have been reported. This case-control study was designed to investigate whether there was an association between phthalate metabolites and childhood obesity in China. A total of 240 pairs of obese/overweight children and age- (±3 months) and gender-matched controls were recruited. Nine phthalate metabolites were analyzed in the first morning urine sample. Physical activity and dietary intake were recorded using validated questionnaires. In monofactor analysis, the levels of monomethyl phthalate (MMP) and monobutyl phthalate (MnBP) in controls were significantly higher than those of overweight/obese children (p < 0.05). Moderate physical activity (p = 0.004), consumption of vegetables, fruits, and tonic were significantly higher in controls (all p < 0.05), and consumption of fried food, western fast food, carbonated drinks, and juice were higher in cases (all p < 0.05). After adjusting for physical activity and dietary intake, neither MMP [OR = 0.825, (95% CI: 0.559-1.217)] nor MnBP [(OR = 0.808, 95% CI: 0.556-1.176)], were significantly associated with obesity. In all models, moderate physical activity was negatively associated and high glucose high fat dietary patterns were positively associated with the risk of childhood obesity (p < 0.01). Diet and physical activity, but not phthalate metabolites were associated with childhood obesity. Further studies are needed to verify our findings. The trial was registered at clinicaltrials.gov as NCT05622513.


Genomic profile of Pancoast syndrome due to hepatocellular carcinoma: A case report.
Hepatocellular carcinoma (HCC) is a common cancer and is frequently diagnosed at a late and unresectable stage with limited effective treatment options. Here, we present the fifth reported case of a 77 year-old male with metastatic HCC presenting as a symptomatic superior sulcus lung tumor and discuss the genomic profile of this rare presentation of HCC for the first time, which included multiple classic mutations in HCC such as TERT, TP53, and WNT/β-catenin signaling as well as in the DNA repair gene ATM. The patient was treated with palliative radiotherapy to the Pancoast tumor followed by atezolizumab plus bevacizumab and passed away 6 months after diagnosis. This rare case highlights the need for effective treatment in aggressive and unresectable HCC and the utility of early genomic studies to allow for targeted therapy such as poly (ADP-ribose) polymerase (PARP)-inhibitors.


Graphene and tricobalt tetraoxide nanoparticles based biosensor for electrochemical glutamate sensing.
An amperometric biosensor based on tricobalt tetraoxide nanoparticles (Co3O4), graphene (GR), and chitosan (CS) nanocomposite modified glassy carbon electrode (GCE) for sensitive determination of glutamate was fabricated. Scanning electron microscopy was implemented to characterize morphology of the nanocomposite. The biosensor showed optimum response within 25 s at pH 7.5 and 37 °C, at +0.70 V. The linear working range of biosensor for glutamate was from 4.0 × 10-6 to 6.0 × 10-4 M with a detection limit of 2.0 × 10-6 M and sensitivity of 0.73 μA/mM or 7.37 μA/mMcm2. The relatively low Michaelis-Menten constant (1.09 mM) suggested enhanced enzyme affinity to glutamate. The glutamate biosensor lost 45% of its initial activity after three weeks.


Regulation in the targeting of TRAIL receptor 1 to cell surface via GODZ for TRAIL sensitivity in tumor cells.
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors, TRAIL-R1 (DR4) and TRAIL-R2 (DR5), promote the selective clearing of various malignancies by inducing apoptosis, holding the promise as a potent therapeutic agent for anticancer. Though DR4 and DR5 have high sequence similarity, differential regulation of both receptors in human tumor cells remains largely unexplored. Here, we repot that golgi-specific Asp-His-His-Cys (DHHC) zinc finger protein (GODZ) regulates TRAIL/DR4-mediated apoptosis. Using the SOS protein recruitment-yeast two-hybrid screening, we isolated GODZ that interacted with the death domain of DR4. GODZ binds to DR4, but not to DR5, through the DHHC and the C-terminal transmembrane domain. Expression level of GODZ affects apoptosis of tumor cells triggered by TRAIL, but not that induced by TNF-α/cycloheximide (CHX) or DNA-damaging drugs. In parallel, GODZ functions to localize DR4 to the plasma membrane (PM) via DHHC motif. Also, introduction of mutation into the cysteine-rich motif of DR4 results in its mistargeting and attenuates TRAIL- or GODZ-mediated apoptosis. Interestingly, GODZ expression is highly downregulated in Hep-3B tumor cells, which show resistance to TRAIL. However, reconstitution of GODZ expression enhances the targeting of DR4 to cell surface and sensitizes Hep-3B cells to TRAIL. Taken together, these data establish that GODZ is a novel DR4-selective regulator responsible for targeting of DR4 to the PM, and thereby for TRAIL-induced apoptosis.


Redirecting photosynthetic electron flux in the cyanobacterium Synechocystis sp. PCC 6803 by the deletion of flavodiiron protein Flv3.
Oxygen-evolving photoautotrophic organisms, like cyanobacteria, protect their photosynthetic machinery by a number of regulatory mechanisms, including alternative electron transfer pathways. Despite the importance in modulating the electron flux distribution between the photosystems, alternative electron transfer routes may compete with the solar-driven production of CO2-derived target chemicals in biotechnological systems under development. This work focused on engineered cyanobacterial Synechocystis sp. PCC 6803 strains, to explore possibilities to rescue excited electrons that would normally be lost to molecular oxygen by an alternative acceptor flavodiiron protein Flv1/3-an enzyme that is natively associated with transfer of electrons from PSI to O2, as part of an acclimation strategy towards varying environmental conditions. The effects of Flv1/3 inactivation by flv3 deletion were studied in respect to three alternative end-products, sucrose, polyhydroxybutyrate and glycogen, while the photosynthetic gas fluxes were monitored by Membrane Inlet Mass Spectrometry (MIMS) to acquire information on cellular carbon uptake, and the production and consumption of O2. The results demonstrated that a significant proportion of the excited electrons derived from photosynthetic water cleavage was lost to molecular oxygen via Flv1/3 in cells grown under high CO2, especially under high light intensities. In flv3 deletion strains these electrons could be re-routed to increase the relative metabolic flux towards the monitored target products, but the carbon distribution and the overall efficiency were determined by the light conditions and the genetic composition of the respective pathways. At the same time, the total photosynthetic capacity of the Δflv3 strains was systematically reduced, and accompanied by upregulation of oxidative glycolytic metabolism in respect to controls with the native Flv1/3 background. The observed metabolic changes and respective production profiles were proposedly linked with the lack of Flv1/3-mediated electron transfer, and the associated decrease in the intracellular ATP/NADPH ratio, which is bound to affect the metabolic carbon partitioning in the flv3-deficient cells. While the deletion of flv3 could offer a strategy for enhancing the photosynthetic production of desired chemicals in cyanobacteria under specified conditions, the engineered target pathways have to be carefully selected to align with the intracellular redox balance of the cells.


Flexible cystoscopy and retrograde catheterization in renal transplant patients.
Catheterizing the ureteral orifice of a renal transplant patient can be difficult using traditional rigid instruments. This report describes our experience with 4 patients who underwent cystoscopy and attempted retrograde catheterization of a transplanted ureter. In all cases attempted catheterization using rigid instruments was unsuccessful, while in 3 of the 4 patients, successful cannulation was accomplished using the flexible cystoscope. These results are attributed to the expanded range of motion achieved with the tip of the flexible cystoscope. The increased mobility afforded by this instrument facilitates access to the ureteral orifice, which is often positioned on the distensible portion of the bladder.


Results of a randomized double-blind crossover trial of scopolamine versus placebo administered by transdermal patch for the control of cisplatin-induced emesis.
We tested the efficacy of scopolamine delivered by transdermal patch in preventing emesis induced by cisplatin as a single agent or in combination. The administration of scopolamine by transdermal patch resulted in acceptable side effects but was ineffective in controlling cisplatin-induced emesis.


Biometry of cataractous eyes using partial coherence interferometry: clinical feasibility study of a commercial prototype I.
To evaluate the clinical feasibility of the prototype version of a commercial partial coherence interferometry instrument (axial length measurement, ALM, Carl Zeiss Jena) for noninvasive, high-precision biometry in cataractous eyes. Department of Ophthalmology, Vienna General Hospital, and Institute of Medical Physics, University of Vienna, Austria. The preoperative axial length in 49 eyes of 37 cataract patients was measured with the commercial (ALM) and laboratory (PCI) prototypes of the partial coherence interferometry instrument, as well as with immersion ultrasound (IUS). Axial length measurements with the ALM and PCI did not differ significantly (P = .23). Both prototypes assessed longer axial lengths than the IUS technique (P < .0001; median 203 microm; range -476 to +635 microm). The precision of the axial length measurement was 18 microm, 28 microm, and 54 microm with the PCI, ALM, and IUS, respectively. Partial coherence tomography is a high-precision, high-resolution, noncontact biometric technique.The commercial PCI prototype is practical in clinical use, with improved comfort for patients, no need for anesthesia, and a reduced risk of infection. However, the difference between the PCI and IUS in axial length measurement must be considered when using the constants supplied by intraocular lens (IOL) manufacturers for IOL power calculations.


Annexin V binding assay as a tool to measure apoptosis in differentiated neuronal cells.
We describe a rapid and reliable method to quantitate the extent of apoptosis in neuronal cell cultures. Based on their annexin V-affinity, resulting from phosphatidylserine (PS) exposure at the outer leaflet of the plasma membrane, apoptotic cells can be distinguished from annexin V-negative living cells, by using microscopic and flow cytometric procedures. When combined with propidium iodide (PI) the double labeling procedure allows a further distinction of necrotic (annexin V+/PI+), apoptotic (annexin V+/PI-) cells. Furthermore, when the cells are incubated with annexin V prior to harvesting, the former cell populations can be separated from cells damaged during isolation (annexin V-/PI+). In the present paper, we show that the annexin V-binding assay is also applicable to differentiated neuronal cells with fragile neurite outgrowths.


[Mycophenolic acid induced apoptosis in T lymphocytic cells Molt-4 and its mechanism].
To determine whether mycophenolic acid (MPA) exerts an apoptotic effect on human leukemic T cells Molt-4 and to elucidate its possible mechanism. Cell morphology, DNA fragmentation, cell cycle,percentage of annexin V positive cells and enzymatic activity of caspase-3 were measured by microscopic, electrophoretic and flow cytometric techniques respectively. Human leukemic B cell line Raji was used as control. MPA could induce apoptosis of Molt-4 cells with dose-and time-dependent manners; cells were blocked in the S phase. The activity of caspase-3 was enhanced in a dose-dependent manner in Molt-4 cells treated with MPA for 24 h. MPA could not induce apoptosis in Raji cells. MPA can induce apoptosis in Molt-4 cells which may be involved in the activation of caspase-3.


Does early use of enzyme replacement therapy alter the natural history of mucopolysaccharidosis I? Experience in three siblings.
Enzyme replacement therapy is widely used as treatment for mucopolysaccharidosis I (MPS I), and there is evidence that this produces improvement in certain clinical domains. There does appear to be variation in the response of clinical features to treatment once these are established. In a reported sibling pair, when enzyme replacement therapy was commenced pre-symptomatically in the younger child, the natural history of the condition appeared to be affected. We present data from three siblings treated with enzyme replacement therapy at different ages which supports this finding.


Enterprising organizations.
Here's an inside--and detailed--look at how three hospital systems achieved HIMSS Analytics Stage 7, an objective measure of progress toward EMR implementation.


Neighborhood and school ethnic structuring and cultural adaptations among Mexican-origin adolescents.
The ethnic and racial structuring of U.S. neighborhoods may have important implications for developmental competencies during adolescence, including the development of heritage and mainstream cultural orientations. In particular, living in highly concentrated Latino neighborhoods during early adolescence-which channels adolescents into related school environments-may promote retention of the ethnic or heritage culture, but it also may constrain adaptation to the mainstream U.S. culture. We tested these hypotheses longitudinally in a sample of 246 Mexican origin adolescents (50.8% girls) and their parents. Data were collected 4 times over 8 years, with adolescents averaging 12.5 (SD = .58) to 19.6 (SD = .66) years of age across the period of the study. Latino ethnic concentration in early adolescents' neighborhoods promoted the retention of Mexican cultural orientations; Latino ethnic concentration in middle schools undermined the development of mainstream U.S cultural orientations. Findings are discussed in terms of integrating cultural-developmental theory with mainstream neighborhood theory to improve understandings of neighborhood and school ethnic concentration effects on adolescent development. (PsycINFO Database Record


Effectiveness of "Stop the Bleed" Courses: A Systematic Review and Meta-analysis.
Our object was to comprehensively analyze the existing body of evidence to evaluate the Stop the Bleed (STB) course effectiveness and satisfaction and find the direction of improvement for the future. A literature search with the term "Stop the Bleed" in the electronic databases PubMed, Web of Science, EMBASE, Cochrane Library was performed, retrieving records from January 1, 2013 to April 13, 2022 based on Preferred Reporting Items for Systematic Reviews and Meta-Analyses flow diagram. In addition, all selected papers' references were examined for qualified studies that were missed during the first search. Original publications were included that reported on (1) clinical studies of the STB course implementation; and (2) studies comparing students' hemostasis ability and attitude (comfort, confidence, and willingness) before and after the STB course. The literature search and data extraction were done independently by 2 writers. To establish consensus, disagreements will be handled with the help of a third reviewer. For data synthesis, the most inclusive data from studies with repeated data were abstracted. Changes in hemostasis questionnaire scoring and operation evaluation after the STB course were the main outcomes. This systematic review and meta-analysis includes 36 trials with a total of 11,561 trainees. Thirty-one of them were undertaken in the USA, while the other 5, accounting for 13.9%, were conducted in other regions. Among various evaluation methods, 3 trials with 927 trainees indicated that scores of correct uses of tourniquet significantly increased after the STB course (mean difference of post versus pre groups, 44.28; 95% CI 41.24-47.32; p < 0.001). Significant difference was also observed in the willingness to apply a hemostatic dressing in a real-world situation (risk ratio for post versus pre groups, 1.28; 95% CI 1.08-1.52; p = 0.004) (7 studies and 2360 participants). The results indicate that hemostasis knowledge and skills after the STB course had improved, but statistics indicated that STB courses implemented in the USA were more effective than other regions. Meta-analysis showed that comparison before and after the STB course were significantly different. However, the outcome measures in each study were different and could not, therefore, be compiled in all cases. The effectiveness and worth of implementation of STB in different countries should be continuously evaluated in the future.


Mediterranean Diet In Healthy Aging.
The World elderly population is expected to double before 2050. Unhealthy habits and unhealthy lifestyles are commonly associated with age-related diseases or their worsening. Modification in daily lifestyle and diet may help preventing age-related diseases onset and efficiently affecting their evolution, thus promoting the Healthy Aging process, concept recently coined to describe the disease-free aging process. This review highlights the role of nutrition science in promoting healthy aging. Since the Mediterranean Model demonstrated to be a useful style in supporting healthy aging, promotion of this correct lifestyle by health policies seems to be the best approach to achieve this target.


Membrane feeding for infection of the reduviid bug Triatoma infestans with Blastocrithidia triatomae (Trypanosomatidae) and pathogenic effects of the flagellate.
To investigate the pathogenic effects of the trypanosome Blastocrithidia triatomae on Triatoma infestans, a method for direct infection by in vitro feeding was developed. After isolation of B. triatomae cysts from infected dead bugs, a suspension of cysts was disinfected with Desogen and Chloramin T, then mixed with sterile blood, spread on a sterile, parallel-grooved glass plate or a hammered aluminium plate and covered with a thin, sterile silicone membrane. The blood and plate were warmed to 36 degrees-38 degrees C by a heating plate. For investigation of the pathological effects, first instars of T. infestans were allowed to feed through the membrane on a mixture containing 10(6) cyst stages/cm3 blood. Development of about 30% of the third and fourth instar larvae was retarded as compared with that in uninfected control groups, and mortality rates increased in fourth and/or fifth instar larvae. Whereas total mortality rates of about 5% occurred in control groups, about 85% of the larvae died in the infected groups. These effects are very similar to those obtained in previous studies with coprophagic infections.


Proliferation and LDL binding of cultured intimal smooth muscle cells from rabbits.
Cultured intimal smooth muscle cells (SMC) were prepared from rabbits in which a cannula had been inserted into the abdominal aorta 4 weeks previously. The patterns of growth proliferative rates, and lipoprotein metabolism of cultures of intimal and medial SMC from intact aortic media were compared. Intimal SMC proliferated more rapidly than medial SMC. Both low density lipoprotein (LDL) and acetylated LDL bound to intimal SMC, whereas only LDL bound to medial SMC. These findings suggest a phenotypic difference between intimal and medial SMC.


Intermittent hypercapnia-induced phrenic long-term depression is revealed after serotonin receptor blockade with methysergide in anaesthetized rats.
What is the central question of this study? Intermittent hypercapnia is a concomitant feature of breathing disorders. Hypercapnic stimuli evoke a form of respiratory plasticity known as phrenic long-term depression in experimental animals. This study was performed to investigate the putative role of serotonin receptors in the initiation of phrenic long-term depression in anaesthetized rats. What is the main finding and its importance? Phrenic nerve long-term depression was revealed in animals pretreated with the serotonin broad-spectrum antagonist, methysergide. This study highlights that serotonin receptors modulate respiratory plasticity evoked by acute intermittent hypercapnia in anaesthetized rats. This study was performed to test the hypothesis that intermittent hypercapnia can evoke a form of respiratory plasticity known as long-term depression of the phrenic nerve (pLTD) and that 5-HT receptors play a role in the initiation of pLTD. Adult male urethane-anaesthetized, vagotomized, paralysed, mechanically ventilated Sprague-Dawley rats were exposed to an acute intermittent hypercapnia protocol. One group received i.v. injection of the non-selective 5-HT receptor antagonist methysergide and another group received i.v. injection of the selective 5-HT1A receptor antagonist WAY-100635 20 min before exposure to intermittent hypercapnia. A control group received i.v. injection of saline. Peak phrenic nerve activity and respiratory rhythm parameters were analysed at baseline (T0), during each of five hypercapnic episodes, and 15, 30 and 60 min (T60) after the last hypercapnia. Intravenous injection of methysergide before exposure to acute intermittent hypercapnia induced development of amplitude pLTD at T60 (decreased by 46.1 ± 6.9%, P = 0.003). Conversely, in control and WAY-100635-pretreated animals, exposure to acute intermittent hypercapnia did not evoke amplitude pLTD. However, a long-term decrease in phrenic nerve frequency was evoked both in control (42 ± 4 breaths min(-1) at T0 versus 32 ± 5 breaths min(-1) at T60; P = 0.036) and in methysergide-pretreated animals (42 ± 2 breaths min(-1) at T0 versus 32 ± 3 breaths min(-1) at T60; P = 0.028). In WAY-100635 pretreated animals, frequency pLTD was prevented. These results suggest that 5-HT receptors modulate respiratory plasticity induced by acute intermittent hypercapnia in anaesthetized rats.


Temporal variation of methane flux from Xiangxi Bay of the Three Gorges Reservoir.
Three diel field campaigns and one monthly sampling campaign during June 2010-May 2011 were carried out to investigate the CH4 flux across the water-gas interface in Xiangxi Bay of the Three Gorges Reservoir, China. The average CH4 flux was much less than that reported from reservoirs in tropic and temperate regions. The photosynthesis of phytoplankton dominated the diel gas fluxes during alga bloom in spring and summer. The maximum monthly flux occurred in June 2010 and corresponded to the lowest water level. Water temperature, sediment temperature, and TOC did not have significant correlation with the monthly CH4 fluxes. Continuously decreasing hydrostatic pressure and the low water level resulted in more CH4 emission at the sediment-water during the discharging period, and thus increases the CH4 effluxes because the diffusion time through a thin water column is shorter and less CH4 may be oxidized compared with that in a long water column.


Progression to AIDS in relation to clinical factors and clotting product consumption. A 14-year follow-up of a cohort of 52 Dutch HIV-1-infected haemophilic patients.
In order to describe the progression of HIV disease in a cohort of Dutch HIV-1-infected patients with haemophilia and to study the influence of clinical and clotting factors, we performed a 15-year longitudinal clinical and laboratory study. The progression to AIDS in relation to type of haemophilia, antibodies against factor VIII, age, cytomegalovirus (CMV) infection, and type and total amount of clotting factor consumption was studied in 52 haemophilic patients infected with HIV-1 between 1981 and July 1985. The progression to AIDS was faster in the group of patients with antibodies against factor VIII (inhibitors) and in the group of patients with haemophilia B than in the group of patients with haemophilia A without inhibitors. We concluded that HIV-infected patients with haemophilia A with inhibitors and patients with haemophilia B show a significantly faster progression to AIDS than do HIV-infected patients with haemophilia A without inhibitors. These differences are independent of the total amount of clotting factor used and other known risk factors such as age at seroconversion and CMV infection and may be due to the type of clotting product used.


Artificial insemination in canids: a useful tool in breeding and conservation.
Artificial insemination (AI) and semen freezing have become services available to dog owners worldwide, and the demand for services to freeze semen is increasing. In other canids such as the fox, the fur industry utilizes fresh or frozen semen to artificially inseminate vixens to produce pelts. Clearly, AI facilitates the use of a male to sire several females by diluting the ejaculate, increases breeding hygiene, and allows crossing between species with slightly different breeding seasons. The African wild dog (Lycaon pictus) is currently considered by the World Conservation Union (IUCN) as one of most endangered canids. In captive populations of African wild dogs, semen has been frozen with encouraging results, using a standard cryopreservation protocol for domestic dogs, but successful AI has not been reported. In wolves, there is one report regarding the live birth of an offspring after intravaginal AI of a deslorelin-induced estrous female. In 2005, three Mexican gray wolf females were artificially bred by intrauterine insemination with freshly collected semen from unrelated males, and all females whelped. Artificial insemination may be vaginal, intrauterine or intratubal, and the semen may be fresh, fresh and chilled (diluted), or frozen-thawed, and the source of semen may be epididymal or ejaculated. In the domestic dog, the results are good to excellent for AI with all three types of processed semen when the source is ejaculated semen, whereas epididymal sperm still yields poorer results. Species differences in female physiology, as well as differences in the cryotolerance of the sperm from various canid species, warrant further research and development.


Myxoma virus sensitizes cancer cells to gemcitabine and is an effective oncolytic virotherapeutic in models of disseminated pancreatic cancer.
Myxoma virus (MYXV) is a novel oncolytic virus that has been shown to replicate in pancreatic cancer cells, but its efficacy in animal models of pancreatic cancer has not been determined. The efficacy of MYXV as monotherapy or in combination with gemcitabine was evaluated in intraperitoneal dissemination (IPD) models of pancreatic cancer. The effects of an intact immune system on the efficacy of MYXV therapy was tested by comparing immunodeficient versus immunocompetent murine models and combination therapy with gemcitabine was also evaluated. In cell culture, MYXV replication was robust in a broad range of pancreatic cancer cells and also showed increased oncolysis in combination with gemcitabine. In animal models, MYXV treatment conferred survival benefits over control or gemcitabine-treated cohorts regardless of the cell line or animal model used. MYXV monotherapy was most effective in an immunocompetent IPD model, and resulted in 60% long-term survivors. In Pan02 engrafted immunocompetent IPD models, sequential treatment in which MYXV was administered first, followed by gemcitabine, was the most effective and resulted in 100% long-term survivors. MYXV is an effective oncolytic virus for pancreatic cancer and can be combined with gemcitabine to enhance survival, particularly in the presence of an intact host immune system.


Comparison of selected disinfectants efficiency against Listeria monocytogenes biofilm formed on various surfaces.
Listeria monocytogenes is a main etiological factor of listeriosis, spread mainly by food products. In recent years, an increasing number of patients with listeriosis and an augmentation in L. monocytogenes antibiotic resistance, e.g. to penicillin and ampicillin, has been reported. The aim of the study was to characterise the L. monocytogenes strains isolated from fish-processed food products. Species identification, based on the multiplex-PCR reaction, was performed, and the genetic similarity of the isolates was analysed with the RAPD technique. The strains, in the form of planktonic cells and a biofilm, were subjected to drug-susceptibility analysis, and the effect of disinfectants on the bacillus cells was evaluated. All of the analysed strains were of the Listeria monocytogenes species. Three genetically distant strains were detected, i.e. Lm I, Lm II and Lm III. Approximately 66.6% penicillin-resistant and 66.6% cotrimoxazole-resistant strains were found. No erythromycin-resistant strain was detected. The Lm II strain was simultaneously resistant to four antibiotics, i.e. penicillin, ampicillin, meropenem and cotrimoxazole. The strongest biofilm was formed on aluminium foil and the weakest on rubber. The tested disinfectant antibiofilm effectiveness was related to the type of surface. The most effective agent was paracetic acid and hydrogen peroxide (elimination rate 5.10-6.62 log CFU × cm-2 and 5.70-7.39 log CFU × cm-2 after 1- and 5-min exposure, respectively) and the least-sodium hydroxide (elimination rate 0.52-1.20 log CFU × cm-2 and 0.98-1.81 log CFU × cm-2 after 1- and 5-min exposure, respectively). Further studies on a greater number of L. monocytogenes strains are recommended.


Transcriptional analysis of major chaperone genes in salt-tolerant and salt-sensitive mesorhizobia.
Salinity is an important abiotic stress that limits rhizobia-legume symbiosis, affecting plant growth, thus reducing crop productivity. Our aims were to evaluate the tolerance to salinity of native chickpea rhizobia as well as to investigate the expression of chaperone genes groEL, dnaKJ and clpB in both tolerant and sensitive isolates. One hundred and six native chickpea mesorhizobia were screened for salinity tolerance by measuring their growth with 1.5% and 3% NaCl. Most isolates were salt-sensitive, showing a growth below 20% compared to control. An association between salt tolerance and province of origin of the isolates was found. The transcriptional analysis by northern hybridization of chaperone genes was performed using tolerant and sensitive isolates belonging to different Mesorhizobium species. Upon salt shock, most isolates revealed a slight increase in the expression of the dnaK gene, whereas the groESL and clpB expression was unchanged or slightly repressed. No clear relationship was found between the chaperone genes induction and the level of salt tolerance of the isolates. This is the first report on transcriptional analysis of the major chaperones genes in chickpea mesorhizobia under salinity, which may contribute to a better understanding of the mechanisms that influence rhizobia salt tolerance.


Hysteroscopic management of cornual ectopic pregnancy.
Cornual ectopic pregnancies have traditionally been treated with systemic methotrexate, cornual resection, or hysterectomy. A 36-year-old newly gravida presented with an 8-week history of amenorrhea and a positive home pregnancy test. A transabdominal sonogram revealed a left cornual ectopic pregnancy. The patient was treated with multiple methotrexate doses, but the gestational sac persisted. Through the hysteroscope, the sac was ruptured, and the placental tissue was removed from the left cornu under sonographic guidance. Two weeks postoperatively, the patient's beta-human chorionic gonadotropin level was negative, and she had a normal pelvic examination and sonogram. Hysteroscopic removal under sonographic guidance after methotrexate treatment is a conservative option for the treatment of cornual ectopic pregnancy in some patients.


Hypothalamic AMPKα2 regulates liver energy metabolism in rainbow trout through vagal innervation.
Hypothalamic AMPK plays a major role in the regulation of whole body metabolism and energy balance. Present evidence has demonstrated that this canonical mechanism is evolutionarily conserved. Thus, recent data demonstrated that inhibition of AMPKα2 in fish hypothalamus led to decreased food intake and liver capacity to use and synthesize glucose, lipids, and amino acids. We hypothesize that a signal of abundance of nutrients from the hypothalamus controls hepatic metabolism. The vagus nerve is the most important link between the brain and the liver. We therefore examined in the present study whether surgical transection of the vagus nerve in rainbow trout is sufficient to alter the effect in liver of central inhibition of AMPKα2. Thus, we vagotomized (VGX) or not (Sham) rainbow trout and then intracerebroventricularly administered adenoviral vectors tagged with green fluorescent protein alone or linked to a dominant negative isoform of AMPKα2. The inhibition of AMPKα2 led to reduced food intake in parallel with changes in the mRNA abundance of hypothalamic neuropeptides [neuropeptide Y (npy), agouti-related protein 1 (agrp1), and cocaine- and amphetamine-related transcript (cartpt)] involved in food intake regulation. Central inhibition of AMPKα2 resulted in the liver having decreased capacity to use and synthesize glucose, lipids, and amino acids. Notably, these effects mostly disappeared in VGX fish. These results support the idea that autonomic nervous system actions mediate the actions of hypothalamic AMPKα2 on liver metabolism. Importantly, this evidence indicates that the well-established role of hypothalamic AMPK in energy balance is a canonical evolutionarily preserved mechanism that is also present in the fish lineage.


Differences in dihydropyrimidine dehydrogenase activities between gastric and colorectal cancer.
We studied the relation between clinicopathological factors and dihydropyrimidine dehydrogenase (DPD) activity in gastric and colorectal carcinomas. Specimens obtained by surgery from 27 gastric and 17 colorectal carcinomas and their normal mucosa were examined. The levels of DPD activity in the gastric carcinomas and their normal mucosa were significantly higher than those in colorectal carcinomas and their normal counterparts, respectively (both P's < 0.0001). The gastric carcinomas had significantly higher DPD activities than their normal mucosa (P = 0.028), but the colorectal carcinomas did not. Among the clinicopathological factors, which included invasion/metastatic status and staging, the only effect was that of the histological differences of gastric cancer on DPD activity. That is, the level of DPD activity of the histologically undifferentiated gastric carcinoma was significantly higher than that of the differentiated type. No prognostic predictive values of DPD were recognized in either gastric and colorectal cancer. In conclusion, the higher DPD activity in gastric cancer than colorectal cancer may be due to the higher DPD activity in the background mucosa of origin, and the higher population of undifferentiated type of histological classification, compared to the colorectal counterparts.


Nanoprobes for in vitro diagnostics of cancer and infectious diseases.
The successful and explosive development of nanotechnology is significantly impacting the fields of biology and medicine. Among the spectacular developments of nanobiotechnology, interest has grown in the use of nanomaterials as nanoprobes for bioanalysis and diagnosis. Herein, we review state-of-the-art nanomaterial-based probes and discuss their applications in in vitro diagnostics (IVD) and challenges in bringing these fields together. Major classes of nanoprobes include quantum dots (QDs), plasmonic nanoparticles, magnetic nanoparticles, nanotubes, nanowires, and multifunctional nanomaterials. With the advantages of high volume/surface ratio, surface tailorability, multifunctionality, and intrinsic properties, nanoprobes have tremendous applications in the areas of biomarker discovery, diagnostics of infectious diseases, and cancer detection. The distinguishing features of nanoprobes for in vitro use, such as harmlessness, ultrasensitivity, multiplicity, and point-of-care use, will bring a bright future of nanodiagnosis.


Effect of indomethacin on adenosine-induced bronchoconstriction.
The exact mechanism of adenosine-induced bronchoconstriction in patients with asthma is unknown. Adenosine contraction of guinea pig trachea was antagonized by inhibitors of cyclooxygenase. The aim of this study was to investigate the effect of indomethacin (100 mg/day) on adenosine-induced bronchoconstriction in 14 asymptomatic patients with asthma. Airway response was evaluated as FEV1, and adenosine was administered as an aerosol diluted in 0.9% saline to produce a concentration range of 0.125 to 4 mg/ml. The dose of adenosine producing a 20% change in FEV1 (PD20) was calculated from the individual semilogarithmic dose-response curve; the results of PD20 were converted to log values for statistical analysis (Student's paired t test). The study was performed on 3 separate days. On the first day, the adenosine challenge was performed, and on subsequent days patients were pretreated with either placebo or indomethacin in a randomized, double-blind manner. Inhaled adenosine caused bronchoconstriction with a geometric mean PD20 of 0.71 mg (95% confidence limits, 0.44 to 1.16). After placebo, a geometric mean PD20 of 0.91 mg (95% confidence limits, 0.53 to 1.58) was obtained. Indomethacin pretreatment decreased adenosine hyperresponsiveness and shifted the dose-response curves of adenosine challenge to the right with a geometric mean PD20 of 1.28 mg (95% confidence limits, 0.64 to 2.56). The effect of indomethacin on adenosine bronchoconstriction (p less than 0.01 versus baseline; p less than 0.05 versus placebo) suggests an indirect mechanism of adenosine on inducing release of arachidonic acid derivatives. Inflammatory mediators inhibited by indomethacin may be involved in adenosine bronchoconstriction, even if this mechanism is not relevant.(ABSTRACT TRUNCATED AT 250 WORDS)


[How Histone Deacetylase 3 Controls Hepcidin Expression and Hepatitis C Virus Replication].
The key role of histone deacetylases (HDAC) in the regulation of the cellular response to infection with the hepatitis C virus (HCV) was first demonstrated in 2008. When studying the metabolism of iron in the liver tissues of patients with chronic hepatitis C, the authors found that the expression of the hepcidin gene (HAMP), a hormone regulator of iron export, is markedly reduced in hepatocytes under conditions of oxidative stress caused by viral infection. HDAC were involved in the regulation of hepcidin expression through the control of acetylation level of histones and transcription factors, primarily STAT3, associated with the HAMP promoter. The purpose of this review was to summarize current data on the functioning of the HCV-HDAC3-STAT3-HAMP regulatory circuit as an example of a well-characterized interaction between the virus and the epigenetic apparatus of the host cell.