Results of a phase I trial of sorafenib (BAY 43-9006) in combination with oxaliplatin in patients with refractory solid tumors, including colorectal cancer.
Sorafenib (BAY 43-9006), a multiple kinase inhibitor, has been shown to inhibit tumor growth and tumor angiogenesis by targeting Raf kinase, vascular endothelial growth factor receptor, and platelet-derived growth factor receptor. In phase I studies, sorafenib demonstrated single-agent activity in patients with advanced solid tumors and was successfully combined with oxaliplatin in preclinical studies. This phase I study investigated the safety, pharmacokinetics, and efficacy of sorafenib in combination with oxaliplatin. Twenty-seven patients with refractory solid tumors were enrolled in the initial dose-escalation part (cohorts 1, 2A, and 2B) and 10 additional patients with oxaliplatin-refractory colorectal cancer were subsequently enrolled in an extension part (cohort 3). Oxaliplatin 130 mg/m2 was given on day 1 of a 3-week cycle and oral sorafenib was administered continuously from day 4 of cycle 1 at 200 mg twice daily (cohort 1) or 400 mg twice daily (cohorts 2A, 2B, and 3). Adverse events were generally mild to moderate and the maximum tolerated dose was not reached. Common adverse events were diarrhea (52% of patients in the dose-escalation part and 20% in the extension part), sensory neuropathy (44% and 20%), and dermatologic toxicities (41% and 80%). No pharmacokinetic interaction between sorafenib and oxaliplatin was detectable. Two patients with gastric cancer had a partial response. Forty-three percent of patients in cohorts 1 and 2A/B and 78% of patients in cohort 3 exhibited stable disease for >or=10 weeks. Continuous oral sorafenib 400 mg twice daily was safely combined with oxaliplatin without detectable drug interactions and showed preliminary antitumor activity in this phase I study. This dose is recommended for phase II studies.


Intradural spinal seeding of a clival chordoma.
Following recent progress in radiological and surgical techniques for treating skull base tumours, more attention should be paid to the relatively rare phenomenon of intradural spinal seeding in cases of clival chordoma. A 53-year-old man harboured intradural spinal seeding from a clival chordoma 11 years after his initial surgical treatment. He had undergone skull base surgery five times with extra- and intradural approaches, and adjuvant radiotherapy. His complaints of lumbago, paraparesis, and sensory disturbance of the lower left extremity prompted us to carry out imaging studies of the spinal cord, which revealed multiple intradural, extramedullary mass lesions at the thoracic and lumbosacral spinal level. The tumours were histopathologically diagnosed as spinal seeding of the clival chordoma with high proliferative potential (Ki-67 labelling index of 8.4%). As long-term local control of skull base chordoma appears to be improving by the introduction of skull base surgical techniques and adjuvant radiotherapy, spinal intradural seeding might emerge as a potential complication. Close observation and timely imaging studies for spinal seeding of skull base chordoma would be required in long-term postoperative follow-ups, particularly for those patients with highly proliferative tumours.


Development of a novel reverse transcriptase polymerase chain reaction to determine the Gram reaction and viability of bacteria in clinical specimens.
To develop a novel RNA based assay to determine the Gram reaction and viability of bacteria in clinical specimens. Reverse transcriptase PCR (RT-PCR) targeting 16SrRNA region was optimized using Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 by using two novel sets of primers. Sixty clinical specimens consisting of 31 intraocular specimens (19 vitreous fluids and 12 aqueous humor), 11 peripheral blood specimens and 18 other clinical specimens were subjected to standard microbiological culture and RT-PCR to determine the Gram reaction and viability of bacteria. The amplified products were subjected to DNA sequencing to identify the bacterium. The sensitivity of RT-PCR was 0.4fg and the primers amplified bacterial cDNA. RT-PCR detected the presence of bacteria in 60 clinical specimens indicating the presence of viable bacteria. Concordant results were obtained with both primer sets. Seventy five bacterium comprising 52 single (69.3%) and 23 mixed bacteria (30.6%), both Gram positive and Gram negative were detected. These results correlated with the bacterial identity by PCR based DNA sequencing. RT-PCR is a reliable tool to identify the presence of viable bacteria and to precisely determine Gram reaction.


Hypoxia induced metabolism dysfunction of rat astrocytes in primary cell cultures.
In order to study the astroglial contribution to hypoxic injury on brain tissue metabolism, modifications of glutamine synthetase (GS) lactate dehydrogenase (LDH) enolase and malate dehydrogenase activity produced by reduced oxygen supply have been determined in primary cultures of astrocytes prepared from newborn rat cerebral cortex. Enzymatic activities were measured immediately after the hypoxic treatment (9 h) and during post injury recovery. GS level is significantly decreased in response to low oxygen pressure and increased above control value during the post hypoxic recovery period. The magnitude of GS reduction by hypoxia depends on the age of the cells in culture. Lactate dehydrogenase and enolase levels were significantly enhanced during the two periods considered. No modification of the MDH level was observed. The synthesis of LDH isoenzymes containing mainly M subunits is specifically induced by hypoxia. Our results suggest that astroglial cells may represent a particularly sensitive target toward hypoxia injury in brain tissue. Low oxygen pressure available may modify some fundamental metabolical functions of these cells such as glutamate turnover and lactic acid accumulation.


Assessment of generalized lymphadenopathy in AIDS research: the degree of clinical agreement.
The degree of clinical agreement amongst different physicians on the presence or absence of generalized lymphadenopathy was assessed in 32 randomly selected participants from a prospective study of male sexual contacts of men with AIDS or an AIDS-related condition (ARC). Three physicians completed the same standard examination that was developed for the main project and conducted the examination of the anatomic regions in the same order on each person, at approximately the same time, and in random order. One physician (Doctor A) was the physician responsible for conducting examinations on the main cohort from which these participants were selected. Intra-observer agreement was assessed by comparing Doctor A's examinations on these participants with those he had recently conducted within a one and a half month period in the main study. Acceptable levels of intra-observer agreement (kappa = 0.72) and interobserver agreement (kappa = 0.66) were demonstrated for the presence or absence of generalized lymphadenopathy for Doctor A and Doctor B, a physician who periodically replaced Doctor A in the main project. Agreement between Doctors A, B, and C, was less satisfactory (kappa of 0.45 and 0.39, respectively). Doctor C was the least experienced with the standardized examination. However, during the progress of this study, agreement between the three doctors improved (kappa values for the latter 16 participants ranged from 0.60 to 0.86) suggesting that experience with the criteria and the standardization of the examination may enhance agreement.(ABSTRACT TRUNCATED AT 250 WORDS)


Nonlinear mediation analysis with high-dimensional mediators whose causal structure is unknown.
With multiple possible mediators on the causal pathway from a treatment to an outcome, we consider the problem of decomposing the effects along multiple possible causal path(s) through each distinct mediator. Under a path-specific effects framework, such fine-grained decompositions necessitate stringent assumptions, such as correctly specifying the causal structure among the mediators, and no unobserved confounding among the mediators. In contrast, interventional direct and indirect effects for multiple mediators can be identified under much weaker conditions, while providing scientifically relevant causal interpretations. Nonetheless, current estimation approaches require (correctly) specifying a model for the joint mediator distribution, which can be difficult when there is a high-dimensional set of possibly continuous and noncontinuous mediators. In this article, we avoid the need to model this distribution, by developing a definition of interventional effects previously suggested for longitudinal mediation. We propose a novel estimation strategy that uses nonparametric estimates of the (counterfactual) mediator distributions. Noncontinuous outcomes can be accommodated using nonlinear outcome models. Estimation proceeds via Monte Carlo integration. The procedure is illustrated using publicly available genomic data to assess the causal effect of a microRNA expression on the 3-month mortality of brain cancer patients that is potentially mediated by expression values of multiple genes.


Emergence of higher levels of invasive and metastatic properties in the drug resistant cancer cell lines after the repeated administration of cisplatin in tumor-bearing mice.
To establish a more suitable model for reflecting biological aggressiveness in clinically recurrent cancers after chemotherapy, we made the in-vitro-established cisplatin-resistant cell lines, by exposing the parental tumor cell lines to cisplatin in a culture system, and also the in-vivo-established cisplatin-resistant cell lines by repeated cisplatin administration to parental tumor-bearing mice. Although both cell lines similarly demonstrated a clinically relevant low level of drug resistance (from 1.5 to 2.9 times more resistance to cisplatin than their parental cell lines), only the in-vivo-established cisplatin-resistant cell lines showed significantly enhanced metastatic properties with a 2.1- to 3.4-fold increase in the number of lung metastatic nodules. These enhanced metastatic properties were caused by tumor invasiveness in combination with various levels of enhancement of cell attachment, proteolytic enzyme activity and cell motility. We concluded that anticancer drugs such as cisplatin could promote tumor progression only in the drug-resistant cell lines established in vivo. As a result, these cell lines are considered to be a more faithful and useful model for expressing biological aggressiveness in clinically recurrent cancers after chemotherapy than the conventional drug-resistant cell lines established in vitro.


Analysis of Saccharides by the Addition of Amino Acids.
In this work, we present the detection sensitivity improvement of electrospray ionization (ESI) mass spectrometry of neutral saccharides in a positive ion mode by the addition of various amino acids. Saccharides of a broad molecular weight range were chosen as the model compounds in the present study. Saccharides provide strong noncovalent interactions with amino acids, and the complex formation enhances the signal intensity and simplifies the mass spectra of saccharides. Polysaccharides provide a polymer-like ESI spectrum with a basic subunit difference between multiply charged chains. The protonated spectra of saccharides are not well identified because of different charge state distributions produced by the same molecules. Depending on the solvent used and other ions or molecules present in the solution, noncovalent interactions with saccharides may occur. These interactions are affected by the addition of amino acids. Amino acids with polar side groups show a strong tendency to interact with saccharides. In particular, serine shows a high tendency to interact with saccharides and significantly improves the detection sensitivity of saccharide compounds. Graphical Abstract .


Colorimetric sensor array combined with chemometric methods for the assessment of aroma produced during the drying of tencha.
The aroma produced during drying is an important indicator of tencha and needs to be monitored. This study constructed an olfactory visualization system for assessing tencha aroma using colorimetric sensor array (CSA) combined with chemometric methods. The 16 chemically responsive dyes were selected to obtain aroma information of tencha samples and extracted image data of aroma information by machine vision algorithms. Subsequently, k-nearest neighbor, support vector machine, classification and regression tree, and random forest (RF), four qualitative models were applied to build the mathematical models. The RF model with nine principal components was preferred, with recognition rate of 100.00% and 91.07% for the training and prediction sets, respectively. The experimental results showed that CSA combined with the RF model can be effectively applied to assess tencha aroma. This study provided a scientific and novel method to maintain the stability of tencha quality in the production process.


Hazards of calcium gluconate therapy in the newborn infant: intra-arterial injection producing intestinal necrosis in rabbit ileum.
Five infants received 10% calcium gluconate via umbilical artery catheters, which resulted in intestinal bleeding and lesions of the buttock, anus, groin, and thigh. The effects of intra-arterial calcium gluconate in two animal models were investigated. Injection of calcium into the aorta in the region of the posterior mesenteric artery resulted in immediate hyperperfusion of the descending colon; this may be an early hemodynamic response to injury in the area of colon supplied by this vessel. Injections into the arterial arcade of the rabbit ileum resulted in intestinal necrosis and villous atrophy. The use of umbilical artery catheters for administration of calcium gluconate is potentially hazardous.


T cell activation in the elderly: evidence for specific deficiencies in T cell/accessory cell interactions.
We studied T cell activation in the healthy aged (greater than 70 years) by examining lymphocyte proliferative responses to various mitogenic stimuli in accessory cell (AC)-dependent and AC-independent systems. Results show that despite a near normal response to the anti-CD3 monoclonal antibody (mAb) OKT3, peripheral blood mononuclear cells (PBM) from the elderly exhibit a profound reduction in phytohaemagglutinin (PHA)-responsiveness (approximately 30% of young adults). This deficit becomes even more severe at suboptimal doses of PHA. Adding exogenous interleukin-2 (IL-2) or pretreating the AC population with gamma interferon (IFN-gamma) returns the level of proliferation to that seen with young adults. Furthermore, replacing "old" AC with AC from young adults or with U937 (a monocytic cell line) in T cell/AC cell-mixing experiments restores PHA-responsiveness in 70% of cases. On the other hand, AC from the aged fully support PHA responses in T cells from young adults. In AC-depleted cultures, purified T cells from the aged respond normally to the co-mitogenic stimuli, PHA + PMA. Taken together, these results suggest that the age-associated diminution in PHA-responsiveness is due, at least in part, to specific deficiencies in T cell/AC communication.


On the meaning of density dependence.
Despite a long history, the term "density dependence" lacks a generally accepted definition. A definition is offered that seems consistent with most other definitions and general usage, that is, a density-dependent factor is any component of the environment whose intensity is correlated with population density and whose action affects survival and reproduction. This definition is used in evaluating the role of territorial behavior, the availability of nest sites, and competition in determining the size of a population. Because neither territory size nor the number of nest sites is correlated with either density or with changes in the birth and death rates of these populations, these cannot be considered density-dependent factors. Competition determines who does breed and who does not rather than the number of breeders, and thus it is not a density-dependent factor determining a population's size.


Protective Effect of Lactobacillus casei on DMH-Induced Colon Carcinogenesis in Mice.
The administration of probiotics is a promising approach to reduce the prevalence of colon cancer, a multifactorial disease, with hereditary factors, as well as environmental lifestyle-related risk factors. Biogenic polyamines, putrescine, spermidine, and spermine are small cationic molecules with great roles in cell proliferation and differentiation as well as regulation of gene expression. Ornithine decarboxylase is the first rate-limiting enzyme for polyamine synthesis, and upregulation of ornithine decarboxylase activity and polyamine metabolism has been associated with abnormal cell proliferation. This paper is focused on studying the protective role of Lactobacillus casei ATCC 393 in a chemically induced mouse model of colon carcinogenesis, directing our attention on aberrant crypt foci as preneoplastic markers, and on polyamine metabolism as a possible key player in carcinogenesis. BALB/c mice were administered 1,2-dimethylhydrazine dihydrochloride (DMH) to induce colon cancer (20 mg/kg body weight, subcutaneous, twice a week for 24 weeks). L. casei ATCC 393 was given orally (106 CFU, twice a week), 2 weeks before DMH administration. Hematoxylin and eosin staining, high-performance liquid chromatography, and Western blotting were used to evaluate aberrant crypt foci, urinary polyamines, and ornithine decarboxylase expression in the colon. The experimental data showed that the preventive administration of L. casei ATCC 393 may delay the onset of cancer as it significantly reduced the number of DMH-induced aberrant crypt foci, the levels of putrescine, and the expression of ornithine decarboxylase. Hence, this probiotic strain has a prospective role in protection against colon carcinogenesis, and its antimutagenic activity may be associated with the maintenance of polyamine metabolism.


Transthoracic esophagectomy: a safe approach to carcinoma of the esophagus.
Transthoracic esophagogastrectomy is a safe operation. Mechanical staplers and a cervical anastomosis have been emphasized to avoid catastrophic consequences of anastomotic leaks in the chest. Transhiatal esophagectomy has been proposed to bring the anastomosis into the neck. It is meant to be a palliative procedure and consequently denies the patient the best chance for surgical cure. The emphasis should be on anastomotic technique and sound principles of surgical oncology. Since 1980, we have performed 104 esophagectomies for carcinoma of the esophagus. We used a left thoracoabdominal incision for distal tumors (64) and the Ivor Lewis technique (40) for more proximal tumors. A two-layer inverting interrupted silk suture technique was used for all anastomoses. More than 90% of the procedures were performed by resident staff. The operative mortality was 2.9% (3 patients). There were no anastomotic leaks. Five patients required between one dilation and three dilations postoperatively. A positive smoking history was present in 83 patients and substantial alcohol use, in 33. Median estimated blood loss was 500 ml, and 60% of patients required no transfusions. Major complications included pneumonia (12 patients) and reexploration for bleeding (2). Minor complications included atelectasis (71 patients), atrial fibrillation (9), ventricular arrhythmias (9), urinary tract infection (3), and wound infection (2). Squamous cancer was present in 31 patients and adenocarcinoma, in 73. Positive lymph node metastases were present in 75%. Anastomotic recurrence was documented in 6 patients. Standard techniques of esophagogastrectomy and a two-layer anastomosis will give excellent results with low mortality and acceptable morbidity.


Modification of peptides and other drugs using lipoamino acids and sugars.
The scientific literature is full of new small molecules and larger peptides identified as potential pharmaceutical agents for a variety of diseases. The majority of these compounds, however, will never progress into the clinic because of poor oral absorption and low metabolic stability. The development of practical, economic, and widely applicable systems to improve the bioavailability of drugs is a highly sought-after goal. The conjugation of a drug with lipid and/or sugar units represents one of the most important strategies being investigated in this new field of drug delivery. This chapter describes one method of introducing lipidic groups to drugs via lipoamino acids and also provides useful procedures for the efficient incorporation of sugar units into drugs, particularly peptide drugs, via solid phase synthesis.


Plasma pharmacokinetics and faecal excretion of ivermectin (Eqvalan paste) and doramectin (Dectomax, 1%) following oral administration in donkeys.
Ivermectin (IVM- Eqvalan paste, 1.87%) and doramectin (DRM-Dectomax 1%) were each administered orally to donkeys at 200 microgkg(-1) bodyweight. Blood and faecal samples were collected at predetermined times over 30 days and plasma pharmacokinetics and faecal excretion determined. Maximum plasma concentrations (C(max)) of IVM (23.6 ngml(-1)) and DRM (33.9 ngml(-1)) were obtained at (t(max)) 19.2 and 24h, respectively. The area under the concentration curve (AUC) of DRM (228.9 ngdayml(-1)) was significantly larger than that of IVM (119.3 ngdayml(-1)) and mean residence time (MRT) was 6.5 days for IVM and 9.1days for DRM. The highest (dry weight) faecal concentrations (9.33 microgg(-1) - IVM, 12.12 microgg(-1) - DRM) were detected at 55.9 and 48.0 h, respectively and each compound was detected (0.05 microgg(-1)) in faeces between 11h and 9 days following oral administration in donkeys.


Zinc sulfate therapy for infertile male with or without varicocelectomy.
One hundred one men were treated for infertility with 440 mg of zinc sulfate daily for sixty days to two years. All subjects demonstrated low seminal plasma zinc concentrations prior to treatment. Patients were divided into two groups: group [Zn], 65 subjects who underwent zinc therapy alone and group [Var/Zn], 36 subjects who received zinc sulfate therapy after varicocelectomy. In both groups seminal zinc levels were significantly higher (P less than 0.05 and P less than 0.01, respectively) two months after the initiation of zinc therapy. There were no significant changes in sperm count or motility for the [Zn] patients. However, group [Var/Zn] showed a significant increase (P less than 0.05) of sperm motility at two months and twelve months after starting zinc sulfate therapy. We separated the [Var/Zn] patients into two sub-divisions: those who impregnated their wives after therapy, and those who remained infertile despite treatment. Only those patients who impregnated their wives showed a significant (P less than 0.05) increase in sperm motility two months after the initiation of zinc therapy. Those patients of the [Var/Zn] group who remained infertile despite treatment showed a significant increase (P less than 0.05) in sperm motility after twelve months of therapy. Of the [Zn] patients 27.7 per cent (18) successfully impregnated their wives, while the [Var/Zn] patients exhibited a pregnancy rate of 50 per cent (18 patients). These data indicate that zinc sulfate therapy for patients with low seminal zinc concentrations may be an effective treatment for infertile patients especially after varicocelectomy.


Association between perfluoroalkyl acids and liver function: Data on sex differences in adolescents.
The data herein presented show multivariate linear regressions performed to examine the association between individual serum perfluoroalkyl acids (PFAAs) [perfluorooctane sulfonic acid (PFOS); linear form of perfluorooctanoic acid (PFOA); perfluorohexane sulfonic acid (PFHxS); perfluorononanoic acid (PFNA)], and biomarkers of liver function (Sex Differences in the association between perfluoroalkyl compounds and liver function in US adolescents: analyses of NHANES 2013-2016). Data relate to male and female adolescents (ages 12-19 years) who participated to the 2013-2016 National Health and Nutrition Examination Survey. The outcome of interest was represented by changes in biomarkers of liver function, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transferase (GGT). Serum measurement values of ALT, AST and GGT were natural log-transformed. Data represent multivariate regression analyses with the single PFAA as β coefficients (and 95%CI), adjusted by age, race/ethnicity, body weight, education, income-to-poverty ratio, and exposure to smoking. Single PFAAs were used as continuous natural log-transformed predicted variables in males and females. Analyses were performed also with individuals PFAAs categorized via sex-specific weighted quartile, with cutoffs based on the weighted distribution of the single PFAA in the study population. Because the dependent variables (ALT, AST and GGT) were log-transformed, data were re-transformed by exponentiation of the β coefficients, and presented as percent differences estimated by comparing each of the upper three quartiles to the lowest quartile using the formula 100*(eβ -1). Together, these data can serve as a basis to analyze associations between liver function and PFAA exposure taking into account sex differences in adolescent populations.


Systemic acute-phase response after laparoscopic and open cholecystectomy.
Cytokines are the main mediators of inflammation and the response to trauma. The purpose of this study was to compare variations in cytokine levels following laparoscopic cholecystectomy (LC) and mini-laparotomy cholecystectomy (OC), since these two types of operations were considered to be a unique model for examining the role of local tissue injury in postoperative inflammatory reactions. A total of 40 patients were studied. Eighteen of them underwent LC; the remaining 22 were operated on using the open technique. Systemic concentrations of interleukin-6 (IL-6), interleukin-1 (IL-1), tumor necrosis factor (TNF), and C-reactive protein (CRP) were measured before and after the operation. In addition, we compared pre- and postoperative white blood cell (WBC) counts, postoperative body temperature, and length of postoperative hospitalization. There was no difference between the two groups in IL-1 and TNF response. The rise in plasma IL-6 levels (18.86 +/- 9.61 vs 5.00 +/- 0.0 pg/ml, p < 0.0001) and CRP (8.40 +/- 5.81 vs 1.43 +/- 1.30 mg/dl, p < 0.001) were more marked after open cholecystectomy than after the laparoscopic procedure. There was no correlation between serum CRP concentrations and the other postoperative parameters. The magnitude of the acute-phase response was less pronounced following laparoscopic cholecystectomy, consistent with a reduction in tissue trauma.


Improving the oxidative stability of fish oil nanoemulsions by co-encapsulation with curcumin and resveratrol.
Tuna fish oil (TFO), is a rich source of omega-3 fatty acids comprising particularly docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), which are essential for the human's health enhancement. However, their foremost problems are poor solubility, low bioavailability, and easy oxidization, which limit their wide range of applications in food and pharmaceutical products. The aim of this research is to develop TFO nanoemulsions encapsulating curcumin (CUR) and resveratrol (RES) as co-antioxidants, stabilized with different concentrations of two modified starches including Purity Gum 2000 (PG) and Purity Gum Ultra (PGU). The effect of diverse quantities of modified starches on droplet size, droplet charges, viscosity, and oxidative stability of produced nanoemulsions was assessed at different storage temperatures (4, 25, and 40 °C). Furthermore, to assess the antioxidant activity and capacity of loaded nanoemulsions, DPPH, and ABTS assays were used, respectively. Among various samples, PGU2.5 and PG9 emulsified by PGU and PG2000 had showed the premium results of stability during storage at all temperatures over other formulations. Even at 40 °C, 199.10 μL/mL of PGU2.5 and 258.59 μL/mL of PG9 were required to attain the level of DPPH IC50, which were the lowest concentration as compare to other nanoemulsions. Taken together, it was accredited that co-encapsulation of CUR and RES inside the TFO nanoemulsion-based delivery systems can be efficient for the production of functional foods.


Wisconsin Card Sorting Test: Normative data and experience.
The Wisconsin Card Sorting Test (WCST) has been increasingly employed as a clinical neuropsychological instrument. However, in India the use of WCST is still in a relatively preliminary stage. To analyse the utility of WCST in the Indian population. Fifty-three subjects in the age group of 20-50 years, comprising both men and women, were recruited for the study. The normality was established by administering the General Health Questionnaire as a screening instrument to evaluate their health status. The WCST was administered and the norms for various dimensions were established; these were compared with those of normal healthy individuals from the West as per the WCST manual. The significance of differences and the experience of administration have been described. The present study found highly significant differences between the means on almost all WCST scores among the Western and the Indian sample, except for the number of correct responses.


Expression of the matrix metalloproteinases 2 and 9 in the rat small intestine during intrauterine and postnatal life.
The expressions of matrix metalloproteinases 2 and 9 have been described during the development, as an example in heart and tooth but not in the small intestine yet. In this context, this study aimed to evaluate the expressions of MMP-2 and MMP-9 in the small intestine of Wistar rats during intrauterine (IU) and postnatal (PN) life. Expressions were determined on the 15th and 18th days of IU life and the 3rd, 10th, 17th, 25th, and 32nd days of PN life. Intestinal samples obtained from six animals were submitted to zymography, immunohistochemistry, and staining with Masson's trichrome. The results showed that MMP-2 and MMP-9 were not expressed during IU life; however, after birth, MMP-9 was immunolocalized in the goblet and mast cells. In conclusion, our results showed that MMP-2 and MMP-9 were not expressed in absorptive epithelial cells during the IU period of the small intestine but after birth, MMP-9 was expressed in the goblet cells, and mast cells present in the lamina propria.


One hundred fold increased activity of Aeromonas aminopeptidase by sequential substitutions with Ni(II) or Cu(II) followed by zinc.
Full substitution of Cu(II) or Ni(II) for the two g-atom zinc in Aeromonas aminopeptidase hyperactivates the enzyme 6.5 and 25 fold respectively. Even greater enhancements of activity can be achieved with mixed metal substitutions. Thus, apoenzyme reactivated by first adding one g-atom zinc followed by one g-atom of either Cu(II) or Ni(II) is 15 and 22 times more active than the native enzyme. Reversing the order, i.e. by first adding either one g-atom Cu(II) or Ni(II) followed by one g-atom zinc, activates the enzyme nearly 100 fold. The order of metal addition is critical and suggests the existence of two non-identical metal sites, each with a different function.


Bone mineral content, corrected for height or bone area, measured by DXA is not reduced in children with chronic renal disease or in hypoparathyroidism.
The combination of poor growth and parathyroid and mineral disorders complicates the diagnosis of renal bone disease in children with chronic renal insufficiency (CRI), and the role of dual X-ray absorptiometry (DXA) is unclear. We aimed to examine the role of DXA in assessing variation in size-adjusted bone mineral content (BMC) in children with CRI and compare it with a cohort with hypoparathyroidism (HPT) and pseudo-hypoparathyroidism (PHPIa). In 29 patients with CRI (21 male) with a median age of 11 years (10th, 90th centiles 4.4, 14.6) and 10 patients with HPT and PHPIa (three male), with a median age of 13.7 years (7, 16) lumbar spine (LS) and total body (TB) BMC were measured by DXA. Age-, gender- and height-matched data allowed calculation of percentage predicted bone area for age and gender (pBAr) and percentage predicted BMC for bone area and height. In the CRI group, the median glomerular filtration rate (GFR) was 27.4 ml/min per 1.73 m2 (7.1, 69.5), and the median duration of illness was 9.3 years (2.1, 12.1). Median height standard deviation score (Ht SDS) was -1.6 (-3.0, 0.3), and, as expected, median LS and TB pBAr were low at 82% (68, 974) and 76% (63, 92), respectively. LS and TB predicted BMC (pBMC) SDS (corrected for bone size) were generally high, with a median value of 0.4 (-0.9, 1.4) and 0.4 (-0.1,0.9), respectively. Analysis of the prepubertal subset of children (n=15) showed that median percentage predicted LS BMC for height was 104% (80, 116), whereas the median TB BMC for height was 96% (74, 108). Median Ht SDS of the HPT and PHPIa cohort was -0.3 (-2.9, 0.3) and median LS and TB pBAr were 90% (66, 100) and 91% (76, 98), respectively. Median LS and TB pBMC SDS were 0.6 (-0.4, 1.8) and 0.7 (0.3, 1.1), respectively. Median percentage predicted LS and TB BMC for height were 102% (82, 114) and 102% (92, 122). There was no relationship between pBMC SDS and duration of illness, GFR, vitamin D dose, serum intact parathyroid hormone (PTH), serum calcium/phosphate product or serum total alkaline phosphatase (ALP) in the CRI or the HPT cohort. However, one of the highest pBMC SDSs was recorded in a child with PHPIa before she started on any treatment. In children with CRI, BMC, when adjusted for co-existing growth retardation, is similar to that observed in children with hypoparathyroidism. The correct reading of BMC needs a correction for bone size.


The identification of 18 nuclear genes required for the expression of the yeast mitochondrial gene encoding cytochrome c oxidase subunit 1.
Eighteen nuclear mutants of the yeast Saccharomyces cerevisae, each disturbed in the biosynthesis of the mitochondrially encoded cytochrome c oxidase subunit 1 (cox1) and each representing a distinct complementation group, have been examined to identify the level at which COX1 expression is affected. RNA blotting revealed that most have a defect in the processing of COX1 precursor-mRNA; only a few are defective in COX1 transcription and/or pre-mRNA stability. In most RNA-processing mutants, the absence of the COX1 messenger results from a defect in the splicing of one or more COX1 introns. In turn, this defect can be ascribed to a mutation in a nuclear gene which is either directly involved in splicing or else acts indirectly by impairing COX1 translation.


Thyroid dysfunction from inhibitor of fibroblast growth factor receptor.
Thyroid dysfunction has not been previously reported in clinical trials of selective fibroblast growth factor receptor (FGFR) inhibitors including AZD4547. Herein, we report a case of worsening hypothyroidism in a patient with advanced urothelial cancer treated with AZD4547. An 80-year-old Caucasian female with metastatic urothelial carcinoma failed first-line chemotherapy with gemcitabine and carboplatin and second-line treatment with atezolizumab, an inhibitor of programmed cell death ligand 1. She developed hypothyroidism at completion of atezolizumab treatment and responded to levothyroxine. Subsequently she was enrolled to a phase II study and received AZD4547 due to an actionable mutation at FGFR3 found in tumor biopsy. Two months later, she experienced recurrent hypothyroidism symptoms, and was hospitalized twice for small bowel obstruction. Her thyroid stimulating hormone level was significantly increased to 2957 uIU/mL (reference range 0.8-7.7 uIU/mL). Her levothyroxine dose was increased accordingly. Her thyroid function returned to normal 1 month afterwards, and small bowel obstruction did not recur. Further reports and studies will be needed to confirm the relationship between AZD4547 and hypothyroidism. Based on this observation and possible mechanisms for thyroid dysfunction discussed in this paper, routine thyroid function monitoring in patients receiving FGFR inhibitor should be considered.


Paracellular fluid transport by epithelia.
The evidence that a major fraction of water crosses the paracellular route during isotonic fluid transfer is reviewed together with a description of the theory and experimental results derived from extracellular probe studies. Four transporting epithelia which have been studied using the method are gallbladder, intestine, Malpighian tubule, and salivary gland. It is concluded that paracellular probe flows are not due to simple convection generated by osmotic flow through the junctions but are generated by active fluid transport within the junction: a mechano-osmotic process. The geometry of the pathway involved would indicate that some salt accompanies the paracellular fluid, representing a hypo-osmotic flow. Transport of salt by the cell route, which may be accompanied by some water, represents a hypertonic flow. The problem then becomes one of balancing the two to produce an isotonic transportate. We suggest, using recent data from knockout mice, that some aquaporins are functioning in different epithelial tissues as osmo-comparators within a feedback loop that regulates the paracellular fluid flow rate. This results in an overall quasi-isotonic transport by the epithelium. The model is applied to forward-facing systems such as proximal tubule and backward-facing systems such as exocrine glands.


Synthesis of sulfonamide, amide and amine hybrid pharmacophore, an entry of new class of carbonic anhydrase II inhibitors and evaluation of chemo-informatics and binding analysis.
Selective inhibition of carbonic anhydrase (CA) enzyme is an active area of research for medicinal chemists. In the current account, a hybrid pharmacophore approach was employed to design sulfonamide, amide and amine containing new series of potent carbonic anhydrase II inhibitors. The aromatic fragment associated with pharmacophore was altered suitably in order to find effective inhibitors of CA-II. All the derivatives 4a-4m showed better inhibition compared to the standard acetazolamide. In particular, compound 4l exhibited significant inhibition with IC50 value of 0.01796 ± 0.00036 µM. The chemo-informatics analysis justified that all the designed compounds possess <10 HBA and <5 HBD. The ligands-protein binding analyses showed that 4l confined in the active binding pocket with three hydrogen bonds observed with His63, Asn66 and Thr197 residues.


Effects of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in zebrafish: general and reproductive toxicity.
Mixed-sex populations of young adult zebrafish (~2-month-old) were exposed to measured RDX concentrations of 0, 1 or 9.6 ppm for up to 12 weeks followed by a 15-day rearing period in untreated water. RDX caused high mortality at 9.6 ppm, with most deaths occurring within the first 8 weeks of exposure. RDX at 9.6 ppm caused lower body weights at 4 and 8 weeks of exposure; and at 1 ppm, lower body weight was observed only at 4 weeks. Fish length was not affected by treatment at any time during the exposure period. The bioconcentration factor for RDX seemed to be influenced by time of exposure but not by water RDX concentration; its overall values were 1.01+/-0.13, 0.91+/-0.06 and 2.23+/-0.04 at 4, 8 weeks and 12 weeks, respectively. RDX was not detected in fish collected after the 15-day recovery period. In a separate experiment, adult females and males were separately exposed to RDX at measured concentrations of 0, 0.5 and 3.2 ppm for a period of 6 weeks. Reproductive performance was evaluated by biweekly breeding of the fish and measuring packed-egg volume (PEV) as index of fecundity. At 0.5 ppm, RDX caused elevated PEV levels relative to the control value at 2 weeks but not at 4 or 6 weeks, whereas no significant effects were noted at 3.2 ppm. Egg fertilization and embryo hatching rates were not affected by RDX at any of the concentrations tested. In conclusion, RDX at sublethal concentrations causes short-term negative effects on growth and, at 0.5 ppm, positive effects on fecundity.


Anterior Shoulder Instability in the Professional Athlete: Return to Competition, Time to Return, and Career Length.
Anterior shoulder instability is a common condition in professional athletes, yet little is known about the success of surgery. Return to competition (RTC) is a metric indicative of a successful outcome for professional athletes who undergo anterior shoulder stabilization surgery. To determine the rate of RTC, time to RTC, recurrence rate, and length of career after surgery in professional athletes who had undergone surgical treatment for anterior shoulder instability. Case series; Level of evidence, 4. We evaluated professional athletes who underwent surgical treatment for anterior shoulder instability by a single surgeon between 2007 and 2018. Data from patients' medical records, a patient data registry, basic search engines, sports websites, and individual team websites were used to determine length of professional play before injury, duration of career after surgery, and RTC level. Overall, 23 professional athletes (25 shoulders from 12 contact and 13 noncontact athletes) were identified. The mean age at the time of surgery was 24.3 ± 4.9 years (range, 16-35 years). Primary procedures included arthroscopic Bankart repair (76%; 19/25), open Latarjet (20%; 5/25), and bony Bankart repair (4%; 1/25). Of the 23 athletes, 22 returned to their previous level of competition (96%; 95% CI, 78%-100%). The mean time between surgery and RTC was 4.5 months (range, 3-8 months). There was no difference in time to RTC between contact and noncontact athletes (4.1 vs 4.4 months). There was no difference in RTC rates and time to return for players who received a Bankart repair versus a Latarjet procedure (4.6 vs 4.2 months). A total of 12 participants were still actively engaged in their respective sport at an average of 4.3 years since surgery, while 11 athletes went on to retire at an average of 4.8 years. Duration of play after surgery was 3.8 years for contact athletes and 5.8 years for noncontact athletes (P > .05). In this series, professional athletes who underwent surgical shoulder stabilization for the treatment of anterior glenohumeral instability returned to their presurgical levels of competition at a high rate. No differences in RTC rate or time to RTC were observed for contact versus noncontact athletes or for those who received arthroscopic Bankart repair versus open Latarjet. However, contact athletes had shorter careers after surgery than did noncontact athletes.


Lactoferrin gene polymorphisms associated with clinical mastitis in Honduran Holstein inheritance cows.
Lactoferrin (LTF) is an iron-binding glycoprotein found in milk and other exocrine secretion with antibacterial activity proposed as an alternative to mastitis treatment or prevention. LTF has been proposed as a candidate gene for mastitis resistance selection. The aim of this paper was to assess LTF promotor to explore variations with potential association to mastitis resistance in dairy cows from Honduras. A resequencing of promotor and Exon I of LTF gene in extreme mastitis susceptibility cows (126 Holstein and Holstein crossbred) was performed. Eight polymorphisms were found in promotor region, four of them were novel variations. Two were important by frequency among extreme groups, but a polymorphism in - 421 A/T position was significantly (P = 0.0188) associated to mastitis susceptibility. Results support the key role of regulatory region of LTF gene. Some candidate genes are proposed in association with mastitis traits and implications are discussed.


Body temperature patterns before, during, and after semi-natural hibernation in the European ground squirrel.
Ground squirrels undergo extreme body temperature fluctuations during hibernation. The effect of low body temperatures on the mammalian circadian system is still under debate. Using implanted temperature loggers, we recorded body temperature patterns in European ground squirrels kept in an enclosure under natural conditions. Although hibernation onset was delayed, hibernation end corresponded closely to that measured in a field population. Circadian body temperature fluctuations were not detected during deep torpor, but indications of circadian timing of arousal episodes at higher temperatures were found at the beginning and end of hibernation. One male exhibited synchronised arousals to a relatively constant phase of the day throughout hibernation. All animals first entered torpor in the afternoon. Daily body temperature fluctuations were decreased or distorted during the first days after hibernation. We hypothesise that hibernation may affect the circadian system by either decreasing the expression of the circadian oscillator, or by decreasing the amplitude of the circadian oscillator itself. possibly due to gradual, temperature dependent, internal desynchronisation. The latter mechanism may be beneficial because it might facilitate post-hibernation re-entrainment rates.


Simulation of the stomatal conductance of winter wheat in response to light, temperature and CO2 changes.
The stomata are a key channel of the water cycle in ecosystems, and are constrained by both physiological and environmental elements. The aim of this study was to parameterize stomatal conductance by extending a previous empirical model and a revised Ball-Berry model. Light and CO(2) responses of stomatal conductance and photosynthesis of winter wheat in the North China Plain were investigated under ambient and free-air CO(2) enrichment conditions. The photosynthetic photon flux density and CO(2) concentration ranged from 0 to 2000 micro mol m(-2) s(-1) and from 0 to 1400 micro mol mol(-1), respectively. The model was validated with data from a light, temperature and CO(2) response experiment. By using previously published hyperbolic equations of photosynthetic responses to light and CO(2), the number of parameters in the model was reduced. These response curves were observed diurnally with large variations of temperature and vapour pressure deficit. The model interpreted stomatal response under wide variations in environmental factors. Most of the model parameters, such as initial photon efficiency and maximum photosynthetic rate (P(max)), have physiological meanings. The model can be expanded to include influences of other physiological elements, such as leaf ageing and nutrient conditions, especially leaf nitrogen content.


Sorbitol metabolism in retina studied in vitro.
A new method is described which allows study of the retinal sorbitol metabolism in vitro. Bovine retinal tissue was incubated in tissue culture medium for 24 h and after this time showed only small morphological changes. The sorbitol content doubled at a glucose level of 5.5 mM compared with freshly prepared noncultured retina. Increasing glucose concentrations led to a gradual increase of the sorbitol content. Supplementation of the culture medium with fructose likewise enhanced the retinal sorbitol accumulation. An aldose reductase inhibitor (ICI 128436; Statil) significantly decreased the sorbitol content.


An fMRI protocol for administering liquid incentives to human participants.
This protocol describes the materials and approaches for administering liquid incentives to human participants during fMRI scanning. We first describe preparation of the liquid solutions (e.g., neutral solution and saltwater) and liquid delivery setups. We then detail steps to connect the setups to the computer-controlled syringe pump in the MRI control room, followed by procedures for testing the syringe pump dispensing a liquid bolus during the task. Description of custom software and required adapters for implementing the liquid setup are included. For complete details on the use and execution of this protocol, please refer to Yee et al. (2021).


Flocculation kinetics and mechanisms of microalgae- and clay-containing suspensions in different microalgal growth phases.
Interplays between microalgae and clay minerals enhance biologically mediated flocculation, thereby affecting the sedimentation and transportation of suspended particulate matter (SPM) in water and benthic environments. This interaction forms larger flocs with a higher settling velocity and enhances SPM sinking. The aim of this study was to investigate the flocculation kinetics of microalgae and clay in suspension and to elucidate the mechanisms associated with such interactions. Standard jar test experiments were conducted using various mixtures of kaolinite and microalgal samples from batch cultures (Chlorella vulgaris) to estimate biologically mediated flocculation kinetics. The organic matter (OM) composition secreted by the microalgae was characterized using a liquid chromatography - organic carbon detection system, and quantitative analysis of transparent exopolymer particles was conducted separately. A two-class flocculation kinetic model, based on the interaction between flocculi and flocs, was also adopted to quantitatively analyze the experimental data from flocculation. Results from the flocculation kinetic tests and OM analyses, in association with other data analyses (i.e., floc size distribution and flocculation kinetic model), showed that flocculation increased with OM concentration during the growth phase (10-20 d). However, on day 23 during the early stationary phase, flocculation kinetics started decreasing and substantially declined on day 30, even though the amount of OM (mainly biopolymers) continued to increase. Our results indicate that an adequate quantity of biopolymers produced by the microalgal cells in the growth phase enhanced floc-to-floc attachment and hence flocculation kinetics. In contrast, an excessive quantity of biopolymers and humic substances in the stationary phase enhanced the formation of polymeric backbone structures and flocculation via scavenging particles but simultaneously increased steric stabilization with the production of a large number of fragmented particles.


Testosterone inhibits bradykinin-induced intracellular calcium kinetics in rat aortic endothelial cells in culture.
Sex steroids have been associated with cardiovascular diseases and the modification of the risk of coronary artery disease (CAD). We cultured aortic endothelial cells from young adult male rats and loaded them with Fura 2 in order to evaluate the direct effects of testosterone on endothelial cells and the probable regulation of bradykinin-induced effects on intracellular calcium ([Ca(2+)](i)) kinetics, effects that are mediated through an increase in intracellular [IP(3)], which in turn stimulates the rapid release of Ca(2+) from ER stores. Our results show that testosterone had no direct effects on [Ca(2+)](i) kinetics, but did block bradykinin-induced increases in intracellular calcium concentration in endothelial cells. This effect was concentration-dependent; the steroid was applied only 30 s before bradykinin application and thus, the effect can be considered nongenomic in origin. Membrane localization of a putative androgen receptor in endothelial cells could be responsible for this effect. In summary, testosterone can modulate the effects induced by activation of membrane-bound bradykinin receptors.


Treatment of fragility ankle fractures using hindfoot nail, systemic review.
With an aging population, the prevalence of fragility ankle fractures is rising. The surgical management of these injuries is challenging and associated with high rates of complications. An extensive literature review (inception of data until September 2019) was undertaken to locate previous studies that have addressed the same topic. Seven studies met the inclusion criteria. The post-operative Olerud and Molander scores were comparable/slightly lower than pre-injury scores. Bony union was achieved in 90.3%-100% of cases. Hindfoot nails, when used to treat fragility ankle fractures, can facilitate early rehabilitation and restoration of function among elderly osteoporotic patients.


The Role of Pythagorean Self-Awareness Intervention in Obstructive Sleep Apnea. A Randomized Controlled Trial.
Obstructive sleep apnea (OSA) is a chronic disease causing daytime sleepiness and poor sleep and life quality. So far, its repercussions on psychological health have been poorly addressed in the available literature. The aim of this study was to investigate the effect of a cognitive-based stress management technique, called the Pythagorean Self-Awareness Intervention (PSAI), in patients with OSA. In this randomized controlled (parallel usual care group) trial with a 1:1 allocation ratio, patients in the intervention group (N = 30, 59.7 ± 9.4 years old, 15 females) received PSAI for 8 weeks along with continuous positive airway pressure (CPAP) treatment, while patients in the control group (N = 30, 58.9 ± 9.3, 22 females) received CPAP alone. The primary endpoint included OSA symptoms. Secondary endpoints were sleepiness, sleep quality, depression-anxiety-stress, and affect. All 30 patients in the intervention group showed 100% compliance with PSAI. There was a statistically significant reduction in OSA symptoms in the intervention group compared to the control group (p = 0.021). With regard to secondary endpoints, there were statistically significant improvements in sleep quality (p = 0.001) and positive (p = 0.001) and negative affects (p < 0.001) in the PSAI group versus controls. No side effects were reported by the patients. PSAI may be useful as a complementary tool for the management of patients suffering from OSA. Larger randomized controlled trials are required to validate the results of this study.


ERRα Regulates OTUB1 Expression to Promote Colorectal Cancer Cell Migration.
Ovarian tumor domain-containing ubiquitin aldehyde binding protein 1 (OTUB1) is overexpressed in many cancers and plays an important role in tumor progression and metastasis. However, the molecular mechanisms underlying OTUB1 overexpression are not clear. In this study, we found that estrogen-related receptor alpha (ERRα, also called NR3B1) binds to OTUB1 promoter and regulates its expression in colorectal cancer. Furthermore, ERRα promoted the migration of CRC cells by inducing vimentin expression via OTUB1. Our data show that OTUB1 is a novel target of ERRα and indicate that ERRα-OTUB1 signaling may play a significant role in CRC metastasis.


Teaching cells new tricks.
The direct conversion of one differentiated cell type into another--a process referred to as transdifferentiation--would be beneficial for producing isogenic (patient's own) cells to replace sick or damaged cells or tissue. Adult stem cells display a broader differentiation potential than anticipated and might contribute to tissues other than those in which they reside. As such, they could be worthy therapeutic agents. Recent advances in transdifferentiation involve nuclear transplantation, manipulation of cell culture conditions, induction of ectopic gene expression and uptake of molecules from cellular extracts. These approaches open the doors to new avenues for engineering isogenic replacement cells. To avoid unpredictable tissue transformation, nuclear reprogramming requires controlled and heritable epigenetic modifications. Considerable efforts remain to unravel the molecular processes underlying nuclear reprogramming and evaluate stable of the changes in reprogrammed cells.


Exploration of the Shared Hub Genes and Biological Mechanism in Osteoporosis and Type 2 Diabetes Mellitus based on Machine Learning.
A substantial amount of evidence suggests a close relationship between osteoporosis (OP) and Type 2 Diabetes Mellitus (T2DM), but the mechanisms involved remain unknown. Therefore, we conducted this study with the aim of screening for hub genes common to both diseases and conducting a preliminary exploration of common regulatory mechanisms. In the present study, we first screened genes significantly associated with OP and T2DM by the univariate logistic regression algorithm. And then, based on cross-analysis and random forest algorithm, we obtained three hub genes (ACAA2, GATAD2A, and VPS35) and validated the critical roles and predictive performance of the three genes in both diseases by differential expression analysis, receiver operating characteristic (ROC) curves, and genome wide association study (GWAS) analysis. Finally, based on gene set enrichment analysis (GSEA) and the construction of the miRNA-mRNA regulatory network, we conducted a preliminary exploration of the co-regulatory mechanisms of three hub genes in two diseases. In conclusion, this study provides promising biomarkers for predicting and treating both diseases and offers novel directions for exploring the common regulatory mechanisms of both diseases.


Wing dimorphism in Gryllus rubens: genetic basis of morph determination and fertility differences between morphs.
The genetic basis of wing morph determination and fertility differences between wing morphs were studied in the wing-dimorphic cricket, Gryllus rubens. Using pair corsses, a significant effect of genotype on morph determination was documented in F3 progeny of field-collected crickets. The effect of genotype was significantly stronger in females than in males. Results are consistent with an earlier study (Zera and Tiebel 1988) showing that wing development is more strongly buffered from environmental variation in females. Segregation patterns were consistent with a polygenic mode of inheritance and provided no evidence for the existence of genes of major effect, maternal effects, or sex linkage. Only a weak morph x sex association was observed. These results contrast those of Walker (1987) where sex-linked loci of major effect on morph determination were identified in crosses between long-wing and short-wing-selected strains of G. rubens. Short-winged female G. rubens began ovipositing earlier and oviposited significantly more eggs than long-winged females during the first 24 days after adult eclosion. The greater reproductive output of the short-winged morph was due entirely to greater oviposition during the first two weeks after adult eclosion. Preliminary results indicate that flight may further accentuate the reduced reproductive output of long-winged versus short-winged females. These data provide the foundation for investigating the endocrine basis of morph-associated fertility variation in G. rubens and its mechanistic relationship with morph determination.


Cyclic nucleotide-dependent protein kinase activity in acinar cells from guinea pig pancreas.
Acinar cells from guinea pig pancreas possess two distinct protein kinase activities. Cyclic GMP-dependent kinase elutes as a single peak on diethylaminoethyl (DEAE)- cellulose chromatography, is not inhibited by protein kinase inhibitor, and has a greater affinity for cyclic GMP (half-maximal activation at 20 nM) than for cyclic AMP (half-maximal activation at 100 nM). Cyclic AMP-dependent kinase elutes as two peaks on DEAE-cellulose chromatography, is inhibited by protein kinase inhibitor, and has a greater affinity for cyclic AMP (half-maximal activation at 20 nM) than for cyclic GMP (half-maximal activation at 7 micrometer). Binding of cyclic 3H-nucleotides to the enzyme preparation was rapid, specific, temperature-dependent, and reversible, and there was a close correlation between the ability of a particular cyclic nucleotide to inhibit binding of cyclic 3H-nucleotide and its ability of a particular cyclic nucleotide to inhibit binding of cyclic H-nucleotide and its ability to activate protein kinase. Binding of cyclic 3H--nucleotide could not be described as a simple bimolecular reaction and native cyclic nucleotides accelerated the dissociation of bound, labeled cyclic nucleotide. Vasoactive intestinal peptide or secretin, each of which increases cellular cyclic AMP, caused endogenous activation of protein kinase and inhibition of cyclic [3H]AMP binding but did not alter bindings of cyclic [3H]GMP or cyclic [3H]AMP.


Levels of agreement between cardiac magnetic resonance and conductance catheter measurements of right ventricular volumes after pulmonary artery banding.
Pressure-volume analysis is the gold standard for quantifying pump function of the right ventricle (RV); however, volume measurements based on a conductive catheter may be imprecise. The reference method for volume assessment is cardiac magnetic resonance (CMR). To determine the levels of agreement between RV volume measurements obtained by cine CMR, phase-contrast CMR (PC CMR), and a conductance catheter in an animal model. CMR was performed in 20 sheep three months after pulmonary artery banding. Ejection fraction (EF), end-diastolic (EDV), end-systolic (ESV), and stroke volumes (SV) were obtained by cine CMR and conductance catheter. Statistically significant differences between cine CMR and conductance catheter derived volume measurements were found for EDV (P < 0.001), ESV (P < 0.05), and SV (P < 0.05). Bland-Altman analysis showed very poor agreement between the two methods: EDV, bias 36.27 mL, agreement of limits 1.96-70.57 mL; ESV, bias 15.33 mL, agreement of limits -6.89-37.55 mL; and SV, bias 20.69 mL, agreement of limits 8.01-49.10 mL. Good agreement was found for SV between cine CMR and PC CMR (bias -7.0 mL, agreement of limits -24.01-9.98 mL), while SV derived from PC CMR measurements showed poor agreement with conductance catheter (bias 27.76 mL, agreement of limits -3.84-59.26 mL). Poor agreement between the conductance catheter and CMR RV volume measurements was found. PC CMR and cine CMR measurements of SV agreed well.


A quantitative study of seasonal foraging by the grass harvesting termite, Trinervitermes geminatus (Wasmann), (Isoptera, Nasutitermitinae) in Southern Guinea savanna, Mokwa, Nigeria.
Trinervitermes geminatus (Wasmann) harvests standing grass tussocks which are cut into pieces and carried back to the nest. During this activity a certain proportion is left on the soil surface as litter whilst some is ingested. Foraging is mainly accomplished during the dry season (mid-October to the end of April) for periods of 2-4 h daily, either early in the morning, in the evening, or occasionally during both morning and evening. The duration and daily pattern of foraging is partly dependent on temperature, with a lower temperature threshold of 20° C and an upper threshold of 35° C below or above which foraging is restricted.The quantity of grass cut down and carried back to the nest by a population of 737 m-2 (3.08 g m-2) was estimated at 60.3 kg ha-1a-1 with approximately 18 kg ha-1a-1 cut and left as litter. Estimated consumption whilst foraging was approximately 20.4 kg ha-1a-1, giving a total consumption of about 81 kg ha-1a-1. Compared with a total estimated grass production of 3157 kg ha-1a-1 and consumption by cattle of 1404 kg ha-1a-1, the quantity of grass removed by T. geminatus, amounting to only 3.1% of the net primary production, did not appear to be economically significant in this locality.


Involvement of 5-HT(2) serotonergic receptors of the nucleus raphe magnus and nucleus reticularis gigantocellularis/paragigantocellularis complex neural networks in the antinociceptive phenomenon that follows the post-ictal immobility syndrome.
The post-ictal immobility syndrome is followed by a significant increase in the nociceptive thresholds in animals and men. In this interesting post-ictal behavioral response, endogenous opioid peptides-mediated mechanisms, as well as cholinergic-mediated antinociceptive processes, have been suggested. However, considering that many serotonergic descending pathways have been implicated in antinociceptive reactions, the aim of the present work is to investigate the involvement of 5-HT(2)-serotonergic receptor subfamily in the post-ictal antinociception. The analgesia was measured by the tail-flick test in seven or eight Wistar rats per group. Convulsions were followed by statistically significant increase in the tail-flick latencies (TFL), at least for 120 min of the post-ictal period. Male Wistar rats were submitted to stereotaxic surgery for introduction of a guide-cannula in the rhombencephalon, aiming either the nucleus raphe magnus (NRM) or the gigantocellularis complex. In independent groups of animals, these nuclei were neurochemically lesioned with a unilateral microinjection of ibotenic acid (1.0 microg/0.2 microL). The neuronal damage of either the NRM or nucleus reticularis gigantocellularis/paragigantocellularis complex decreased the post-ictal analgesia. Also, in other independent groups, central administration of ritanserin (5.0 microg/0.2 microL) or physiological saline into each of the reticular formation nuclei studied caused a statistically significant decrease in the TFL of seizing animals, as compared to controls, in all post-ictal periods studied. These results indicate that serotonin input-connected neurons of the pontine and medullarly reticular nuclei may be involved in the post-ictal analgesia.


Segregation of the isozymes of flax genotrophs.
The segregation of isozymes of peroxidase and acid phosphatase in progenies of crosses between large (L) and small (S and L6) flax genotrophs has been determined. The peroxidase isozymes segregated as expected on a simple Mendelian model with a dominant and a recessive allele and with the L genotroph being a homozygous dominant. All the peroxidase isozymes which differed segregated together, so the isozymes are controlled by either a single locus or closely linked loci. The acid phosphatase isozymes in the F1 were all L type, but the segregations observed in the F2 were not always consistent with a simple Mendelian model.


Bone motion analysis from dynamic MRI: acquisition and tracking.
For diagnosis, preoperative planning and postoperative guides, an accurate estimate of joint kinematics is required. It is important to acquire joint motion actively with real-time protocols. We bring together MRI developments and new image processing methods in order to automatically extract active bone kinematics from multi-slice real-time dynamic MRI. We introduce a tracking algorithm based on 2D/3D registration and a procedure to validate the technique by using both dynamic and sequential MRI, providing a gold standard bone position measurement. We present our technique for optimizing jointly the tracking method and the acquisition protocol to overcome the trade-off in acquisition time and tracking accuracy. As a case study, we apply this methodology on a human hip joint. The final protocol (bFFE, TR/TE 3.5/1.1 ms, Flip angle 80 degrees , pixel size 4.7 x 2.6 mm, partial Fourier reduction factor of 0.65 in read direction, SENSE acceleration factor of 2, frame rate = 6.7 frames/s) provides sufficient morphological data for bone tracking to be carried out with an accuracy of 3 degrees in terms of joint angle.


Immunometabolism and Its Potential to Improve the Current Limitations of Immunotherapy.
The last century of research in tumor immunology has culminated in the advent of immunotherapy, most notably immune checkpoint inhibitors. These drugs have shown encouraging results across a multitude of malignancies and have shifted the paradigm of cancer treatment. However, no more than 40% of patients treated with these immune checkpoint blockade inhibitors respond. Thus, resistance is a barrier to therapy that remains poorly understood. All cells require energy and biosynthetic precursors for survival, growth, and functioning, where multiple metabolic pathways allow for flexibility in how nutrients are utilized. A defining hallmark of many cancers is altered cellular metabolism, creating an imbalanced demand for nutrients within the tumor microenvironment. Immunometabolism is increasingly understood to be vital to the functions and phenotypes of a myriad of immune cell subsets. In tumors, the high demand for nutrients by the tumor drives competition between tumor cells and infiltrating immune cells, culminating in dysfunctional immune responses. This chapter discusses the recent successes in cancer immunotherapy and highlights challenges to therapy. We also outline the major metabolic processes involved in the generation of an immune response, how this can become dysregulated in the context of the tumor microenvironment, and how this contributes to resistance to immunotherapy. Finally, we explore the potential for targeting immunometabolic pathways to improve immunotherapy, and examine current trials targeting various aspects of metabolism in an attempt to improve the outcomes from immunotherapy.


Oblique medial malleolar osteotomy for the management of talar dome lesions.
The purpose of this retrospective study was to determine the efficacy of the oblique medial malleolar osteotomy for the management of medial talar dome lesions. Arthroscopy remains a viable option for the management of these lesions; however, the central and posteromedial lesions are often difficult to gain access to with arthroscopy and may require open arthrotomy. Fourteen oblique medial malleolar osteotomies were performed for the surgical management of medial talar dome lesions. Subjective data were collected through the process of questionnaire. Objective criteria consisted of radiographic evaluation and retrospective medical record review. Seven of the 14 patients related a history of trauma preoperatively. The oblique medial malleolar osteotomy demonstrated osseous union at an average of 6.6 weeks. There were no delayed unions or nonunions. None of the patients required a second operative procedure. Nine patients reported excellent postoperative results, two patients had good results, two had fair results, and one patient related a poor postoperative outcome with only 50% relief of pain. Patients greater than the age of 30 were found to have less favorable results. Follow-up ranged from 6 to 72 months, with a mean follow-up of 34 months.


A novel mutation in the fibrinogen Aα chain (Gly13Arg, fibrinogen Nanning) causes congenital dysfibrinogenemia associated with defective peptide A release.
Dysfibrinogenemia is characterized by blood coagulation dysfunction induced by an abnormal molecular structure of fibrinogen. Here, we describe a new case. A 32-year-old female was suspected of having dysfibrinogenemia during routine laboratory screening, based on her decreased functional fibrinogen level, normal fibrinogen antigen level, and prolonged thrombin time. We extracted DNA and performed polymerase chain reaction and DNA sequencing to identify genetic mutation. Fibrin polymerization, the kinetics of the fibrinopeptide release, scanning electron microscopy, mass spectrometric analysis, fibrin cross-linking, sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blot were conducted. DNA sequencing identified a heterozygous point mutation, Gly13Arg in Aα chain. Fibrin polymerization was markedly impaired (prolonged lag phase and decreased final turbidity). The rate and extent of fibrinopeptide A release from the patient were abnormal and reduced. The mass spectrometry analysis revealed the presence of mutant fibrinogen chains in the patient's circulation. Electron micrographs revealed abnormal fibrin clots. Fibrin cross-linking was normal. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blot showed no difference. We report a new case with a mutation in the fibrinopeptide A region, AαGly13Arg. These results indicated that the functional abnormalities were related to delayed and defective fibrinopeptide A cleavage and likely impaired thrombin binding.


Lateral habenula dysfunctions in Tm4sf2-/y mice model for neurodevelopmental disorder.
Mutations in the TM4SF2 gene, which encodes TSPAN7, cause a severe form of intellectual disability (ID) often comorbid with autism spectrum disorder (ASD). Recently, we found that TM4SF2 loss in mice affects cognition. Here, we report that Tm4sf2-/y mice, beyond an ID-like phenotype, display altered sociability, increased repetitive behaviors, anhedonic- and depressive-like states. Cognition relies on the integration of information from several brain areas. In this context, the lateral habenula (LHb) is strategically positioned to coordinate the brain regions involved in higher cognitive functions. Furthermore, in Tm4sf2-/y mice we found that LHb neurons present hypoexcitability, aberrant neuronal firing pattern and altered sodium and potassium voltage-gated ion channels function. Interestingly, we also found a reduced expression of voltage-gated sodium channel and a hyperactivity of the PKC-ERK pathway, a well-known modulator of ion channels activity, which might explain the functional phenotype showed by Tm4sf2-/y mice LHb neurons. These findings support Tm4sf2-/y mice as useful in modeling some ASD-like symptoms. Additionally, we can speculate that LHb functional alteration in Tm4sf2-/y mice might play a role in the disease pathophysiology.


Practice pearls in the management of lung cancer in the elderly.
Lung cancer is a disease that afflicts the elderly. It is a leading cause of cancer mortality worldwide. Treatment of lung cancer which was predominantly combination chemotherapy was initially thought to be too toxic for older patients with cancer due to their frail state. However a number of recent studies have shown that this is not necessarily true and many elderly can actually tolerate combination chemotherapy and derive just as much benefit from it as younger patients with lung cancer do. More recently it has been found that a significant proportion of lung cancer patients have tumors that harbor mutations that are targetable by molecularly targeted therapy (MTT). These targeted therapies have a much better tolerated side effect profile, hence have been used in elderly patient with lung cancer with great success. A new generation of drugs called immune checkpoint inhibitors have now come into the fray with exciting results in the second line treatment of lung cancer with a low side effect profile. A key element in deciding whether an elderly patient with lung cancer can tolerate treatment involves a detailed assessment using the comprehensive geriatric assessment (CGA). A number of CGA and clinical factors have also been found to be able to predict chemotherapy associated toxicity. This review of lung cancer in the elderly was part of a lecture on "Practice pearls in the management of lung cancer in the elderly" presented at the SIOG Annual Meeting in Prague in November 2015.


Collateral urethral duplication in the frontal plane: a spectrum of cases.
Collateral urethral duplication in the frontal plane is a rare anomaly. We report 3 cases of this variant of urethral duplication and review the literature. Double stream was the most frequent complaint and associated midline defects were common. Management, including a unique method of surgical intervention, is discussed.


Patients with Type 1 diabetes consuming alcoholic spirits have an increased risk of microvascular complications.
Our aim was to evaluate the effect of the amount of alcohol consumption and the type of beverage on the risk of diabetic nephropathy and severe diabetic retinopathy. The alcohol consumption data were available from 3608 patients with Type 1 diabetes participating in the Finnish Diabetic Nephropathy Study (FinnDiane). We assessed the cross-sectional association between alcohol consumption and diabetic nephropathy as well as retinopathy. Patients were divided into different groups according to the amount of alcohol and the type of beverage they were consuming. In the multivariate analysis, the odds ratio for nephropathy was 1.39 (95% CI 1.05-1.84) for abstainers and 2.44 (95% CI 1.49-3.99) for former users compared with light consumers. The results were similar in retinopathy, with an odds ratio of 1.42 (95% CI 1.11-1.82) for abstainers and 1.73 (95% CI 1.07-2.79) for former users. No difference between light consumers and moderate or heavy consumers was observed. Compared with wine drinkers, men consuming mostly alcoholic spirits had a higher risk of nephropathy with an odds ratio of 2.80 (95% CI 1.15-6.81). In women, there was no difference in the risk of nephropathy between the different beverage types. Alcoholic spirit consumers had a higher risk of retinopathy with an odds ratio of 2.32 (95% CI 1.35-4.00). There was no difference between wine and beer consumers. Alcoholic spirit drinkers carry a higher risk of nephropathy and severe retinopathy compared with wine drinkers. Lifelong abstainers and former users of alcohol have a higher risk of nephropathy and severe retinopathy compared with light consumers.


Polyene antibiotics in assessing significance of antistreptolysin O activity.
Antistreptolysin O activity in serum is due either to antibody or to altered lipoprotein molecules. The latter can be inhibited by performing antistreptolysin tests using a polyene antibiotic such as amphotericin B as diluent.


Molecular identification and phylogenetic analysis of Heterakis gallinae from native chickens in Egypt.
The genomic DNA was extracted from nematode parasites, Heterakis gallinae from specific host, native chickens. The polymerase chain reaction (PCR) was applied to make amplification of the DNA content that aids in the further studies. The primer was used in the PCR reactions that have the following sequences: forward (5'-GTTTCCGTAGGTGAACCTGC-3') and reverse (5'-ATATGCTTAAGTTCAGCGGGT-3'). It gave total amplified fragment markers at 914 bp. The gene sequences obtained from Egyptian H. gallinae was to make phylogenetic relationships of Egyptian H. gallinae with other nematodes on the database of an accession number (AJ876757), showing that it was nearly similar to Heterakis gallinarum.


Chemically induced hepatotoxicity in human stem cell-induced hepatocytes compared with primary hepatocytes and HepG2.
Stem cell-induced hepatocytes (SC-iHeps) have been suggested as a valuable model for evaluating drug toxicology. Here, human-induced pluripotent stem cells (QIA7) and embryonic stem cells (WA01) were differentiated into hepatocytes, and the hepatotoxic effects of acetaminophen (AAP) and aflatoxin B1 (AFB1) were compared with primary hepatocytes (p-Heps) and HepG2. In a cytotoxicity assay, the IC50 of SC-iHeps was similar to that in p-Heps and HepG2 in the AAP groups but different from that in p-Heps of the AFB1 groups. In a multi-parameter assay, phenotypic changes in mitochondrial membrane potential, calcium influx and oxidative stress were similar between QIA7-iHeps and p-Heps following AAP and AFB1 treatment but relatively low in WA01-iHeps and HepG2. Most hepatic functional markers (hepatocyte-specific genes, albumin/urea secretion, and the CYP450 enzyme activity) were decreased in a dose-dependent manner following AAP and AFB1 treatment in SC-iHeps and p-Heps but not in HepG2. Regarding CYP450 inhibition, the cell viability of SC-iHeps and p-Heps was increased by ketoconazole, a CYP3A4 inhibitor. Collectively, SC-iHeps and p-Heps showed similar cytotoxicity and hepatocyte functional effects for AAP and AFB1 compared with HepG2. Therefore, SC-iHeps have phenotypic characteristics and sensitivity to cytotoxic chemicals that are more similar to p-Heps than to HepG2 cells.


[Viability of a Bac. licheniformis 749/c culture and its formation of penicillinase during storage in the lyophilized state].
The amount of the enzyme produced and the colony morphology of Bac. licheniformis 749/C, the penicillinase-producing organism were studied after storage for 3 years in ampoules at a temperature of 4--10 degrees C, in lyophilized form in sodium glutamate, polyvinylpirrolidone, their mixture and horse serum. The highest rate of the culture growth was observed after storage in lyophlized form in sodium glutamate, though the culture was vialable in all other protective media. Two culture types approximately in an equal ratio were observed in the population platings of the culture lyophilized in various protective media.


Quantitative 2-D gel electrophoresis-based expression proteomics of albumin and IgG immunodepleted plasma.
Proteomic analysis of plasma is challenging because of its large dynamic range, which prevents the detection of low abundance proteins. Immunodepletion of high abundance proteins, such as albumin and IgG, has emerged as a favored technology to overcome this problem; however its suitability in quantitative expression proteomics has not yet been adequately addressed. In this study, albumin and IgG immunodepletion was evaluated by ELISAs and the reproducibility of depletion was tested with 2-DGE. Depletion of plasma resulted in removal of 62+/-1.2% of the total protein, 93+/-1.4% of the albumin (0.43 microg/microL, residual), and 94+/-1.5% of the IgG (0.21 microg/microL, residual). These results were confirmed by immunoblotting. Computerized image analysis of 2-D gels using Progenesis SameSpots software revealed an enhancement in the number of visible spots (675-1325), with 10+/-6% inter-gel variability in spot density. LC-ESI-MS/MS identification of newly resolved protein spots further validated the procedure. An innovative application of the software employed led to identification of 11 proteins lost non-specifically during depletion. This study demonstrates the effectiveness of immunodepletion of albumin and IgG in quantitative 2-DGE-based differential analysis of plasma proteins.


A molecular explanation of frequency-dependent selection in Drosophila.
Frequency-dependent selection provides a means for maintaining genetic variability within populations, without incurring a large genetic load. There is a wealth of experimental evidence for the existence of frequency-dependent changes in genotypic fitness among a wide variety of organisms. Examples of traits which have been shown to be subject to frequency-dependent selection include the self-incompatibility alleles of plants, chromosomal rearrangements in Drosophila, visible mutations, enzyme variants and rare-male mating advantage in Drosophila. These experiments have been interpreted in a number of different ways. Principally, frequency dependence of genotype fitness may result from intergenotype facilitation due to the production of biotic residues, or from the differential use of resources by the competing genotypes. However, it has proved extremely difficult to isolate and identify any biotic residue of importance or, alternatively, to understand the manner in which genotypes partition the environment. Thus, the difficulty in the interpretation of experiments which show frequency-dependent selective effects stems largely from our lack of understanding of the exact physiological mechanisms which produce these frequency-dependent effects. The principal aim of this study was to investigate the mechanisms associated with frequency-dependent selection at the amylase locus in Drosophila melanogaster. The excretion of catalytically active amylase enzyme and its effect on food medium composition were correlated with the outcome of intraspecific competition between amylase-deficient and amylase-producing genotypes. Amylase-producing genotypes were shown to excrete enzymatically active amylase protein into the food medium. The excreted amylase causes the external digestion of dietary starch; this accounts for the frequency-dependent increase in the viability of the amylase-deficient mutants in mixed cultures, maintained on a starch-rich diet.


Epicardial implantation of the automatic implantable defibrillator by left subcostal thoracotomy.
The automatic implantable defibrillator is a valuable addition to the management of certain patients with ventricular tachycardia and/or fibrillation. The original technique required placement of two transvenous electrodes and an epicardial electrode via a left thoracotomy. We have successfully developed an all-epicardial approach using a left subcostal thoracotomy. Previous experience with epicardial pacemaker implantation by this approach suggested it would be a good technique for implantation of the automatic defibrillator. Early results with this approach have encouraged its continued use.


Occurrence of indole in cells of extremely halophilic bacteria.
The neutral lipids of several halophilic bacteria contain a non-isoprenoid compound which was identified by its chromatographic and spectroscopic properties as indole.


Can we build synthetic, multicellular systems by controlling developmental signaling in space and time?
Using biological machinery to make new, functional molecules is an exciting area in chemical biology. Complex molecules containing both 'natural' and 'unnatural' components are made by processes ranging from enzymatic catalysis to the combination of molecular biology with chemical tools. Here, we discuss applying this approach to the next level of biological complexity -- building synthetic, functional biotic systems by manipulating biological machinery responsible for development of multicellular organisms. We describe recent advances enabling this approach, including first, recent developmental biology progress unraveling the pathways and molecules involved in development and pattern formation; second, emergence of microfluidic tools for delivering stimuli to a developing organism with exceptional control in space and time; third, the development of molecular and synthetic biology toolsets for redesigning or de novo engineering of signaling networks; and fourth, biological systems that are especially amendable to this approach.


Improved synthesis of 2'-amino-2'-deoxyguanosine and its phosphoramidite.
2'-Amino-2'-deoxynucleosides and oligonucleotides containing them have proven highly effective for an array of biochemical applications. The guanosine analogue and its phosphoramidite derivatives have been accessed previously from 2'-amino-2'-deoxyuridine by transglycosylation, but with limited overall efficiency and convenience. Using simple modifications of known reaction types, we have developed useful protocols to obtain 2'-amino-2'-deoxyguanosine and two of its phosphoramidite derivatives with greater convenience, fewer steps, and higher yields than reported previously. These phosphoramidites provide effective synthons for the incorporation of 2'-amino-2'-deoxyguanosine into oligonucleotides.


A survey of the practice of thoracic epidural analgesia in the United Kingdom.
A postal survey of the practice of thoracic epidural analgesia was sent to 275 hospitals in the United Kingdom. Responses were received from 70% of hospitals. Informed consent is rarely adequately obtained, with only 28% of respondents mentioning even the most common complications. Epidural cannulation is most often (60%) performed following induction of general anaesthesia, rather than in the awake patient. A test dose of local anaesthetic without adrenaline is usual. Neither aspirin nor low-dose heparin are considered a contraindication. The majority of respondents used a combination of bupivacaine with fentanyl (51%) or diamorphine (40%), usually administered by continuous infusion. Drugs were frequently prepared and adjusted by anaesthetic staff. The majority of epidurals (63%) are nursed in intensive care units postoperatively. Properly funded pain management teams, at present unusual, would facilitate ward-based epidural management and release intensive care resource. A central register of epidural complications is required to provide valuable evidence for the optimum practice of thoracic epidural analgesia.


How readers experience characters' decisions.
When people read narratives, they have ample opportunities to encode mental preferences about characters' decisions. In our present project, we examined how readers' preferences for characters' decisions structure their experiences of story outcomes. In Experiment 1, participants read brief stories and explicitly rated which of two potential decisions they thought the characters should make. The actual decision that each character made was either preferred or nonpreferred by readers. By the end of each story, readers learned whether there was a positive or negative outcome to these decisions. Decisions and outcomes either matched (e.g., a preferred decision followed by a positive outcome) or did not match (e.g., a nonpreferred decision followed by a negative outcome). Participants took longer to read outcome sentences when there was a mismatch. In Experiment 2, we replicated this finding with a task that allowed more natural reading. These results provide converging evidence that readers encode responses to characters' decisions and that these responses affect the time course with which they assimilate story outcomes.


Effects of increase in fish oil intake on intestinal eicosanoids and inflammation in a mouse model of colitis.
Inflammatory bowel diseases (IBD) are chronic intestinal inflammatory diseases affecting about 1% of western populations. New eating behaviors might contribute to the global emergence of IBD. Although the immunoregulatory effects of omega-3 fatty acids have been well characterized in vitro, their role in IBD is controversial. The aim of this study was to assess the impact of increased fish oil intake on colonic gene expression, eicosanoid metabolism and development of colitis in a mouse model of IBD. Rag-2 deficient mice were fed fish oil (FO) enriched in omega-3 fatty acids i.e. EPA and DHA or control diet for 4 weeks before colitis induction by adoptive transfer of naïve T cells and maintained in the same diet for 4 additional weeks. Onset of colitis was monitored by colonoscopy and further confirmed by immunological examinations. Whole genome expression profiling was made and eicosanoids were measured by HPLC-MS/MS in colonic samples. A significant reduction of colonic proinflammatory eicosanoids in FO fed mice compared to control was observed. However, neither alteration of colonic gene expression signature nor reduction in IBD scores was observed under FO diet. Thus, increased intake of dietary FO did not prevent experimental colitis.


Ex vivo 3D diffusion tensor imaging and quantification of cardiac laminar structure.
A three-dimensional (3D) diffusion-weighted imaging (DWI) method for measuring cardiac fiber structure at high spatial resolution is presented. The method was applied to the ex vivo reconstruction of the fiber architecture of seven canine hearts. A novel hypothesis-testing method was developed and used to show that distinct populations of secondary and tertiary eigenvalues may be distinguished at reasonable confidence levels (P < or = 0.01) within the canine ventricle. Fiber inclination and sheet angles are reported as a function of transmural depth through the anterior, lateral, and posterior left ventricle (LV) free wall. Within anisotropic regions, two consistent and dominant orientations were identified, supporting published results from histological studies and providing strong evidence that the tertiary eigenvector of the diffusion tensor (DT) defines the sheet normal.


LINC00958: A promising long non-coding RNA related to cancer.
Long non-coding RNAs (lncRNAs), a class of RNA transcripts longer than 200 nucleotides, do not encode proteins; however, they encode small peptides and micropeptides that act as bioactive peptides with notable effects in regulating the progression of malignant tumors, such as lung and colorectal cancers, and affecting patient prognosis. lncRNAs are important intracellular regulators, particularly in tumorigenesis and tumor progression. Long intergenic non-protein coding RNA958 (LINC00958), which has received increasing attention in recent years, is highly expressed in various malignancies, including head and neck squamous cell carcinoma (HNSC), non-small-cell lung cancer (NSCLC), gastric cancer, hepatocellular carcinoma (HCC), colorectal cancer, bladder cancer, and breast cancer. Here, we reviewed the recent studies on LINC00958 as well as its closely related clinical features and functional regulation in cancers. We systematically expounded the molecular mechanisms underlying the biological functions of LINC00958 in inhibiting cell apoptosis and enhancing the chemoradiotherapy resistance of tumor cells. The upregulation of LINC00958 enhances the resistance of tumor cells to radiotherapy and chemotherapy and induces lymphangiogenesis. Moreover, it is involved in tumor glycolytic metabolism, which plays a crucial role in facilitating the proliferation, invasion, and migration of tumor cells. Additionally, analysis of various studies revealed that LINC00958 acts as an endogenous competitive RNA (ceRNA) and regulates the malignant behavior of tumor cells through the miRNA-mRNA axis. Collectively, the use of LINC00958 as a novel biomarker and therapeutic target for the clinical diagnosis and treatment of different cancers has bright prospects in the future.


Simple and sensitive electrogenerated chemiluminescence peptide-based biosensor for detection of matrix metalloproteinase 2 released from living cells.
A simple and sensitive electrogenerated chemiluminescence biosensor was developed to monitor matrix metalloproteinase 2 (MMP-2) by employing a specific peptide (CGPLGVRGK) as a molecular recognition substrate. Bis(2,2'-bipyridine)-4'-methyl-4-carboxybipyridine-ruthenium N-succinimidyl ester-bis(hexafluorophosphate) (Ru(bpy)2(mcbpy-O-Su-ester)(PF6)2 (Ru1) was used as ECL-emitting species and covalently labeled onto the peptide through NH2-containing lysine on the peptide via acylation reaction to form Ru1-peptide as an ECL probe. An ECL peptide-based biosensor was fabricated by self-assembling the ECL probe onto the surface of gold electrode. MMP-2 can specifically cleave the Ru1-peptide on the electrode surface, which led the partly Ru1-peptide to leave the electrode surface and resulted in the decrease of the ECL intensity obtained from the resulted electrode in 0.1 M phosphate-buffered saline (pH 7.4) containing tri-n-propylamine. The decreased ECL intensity was piecewise linear to the concentration of MMP-2 in the range from 1 to 500 ng/mL. Moreover, the ECL biosensor is successfully applied to detection of MMP-2 secreted by living cell, such as HeLa cells. Additionally, the biosensor was also applied to the evaluation of matrix metalloproteinase inhibitors. The strategy presented here is promising for other disease-related matrix metalloproteinase assay and matrix metalloproteinase inhibitor profiling with sensitivity and simplicity. Graphical Abstract Detection of MMP-2 released from living cells by ECL peptide-based biosensor.


[Blood supply and function of the scaphoid bone].
The proximal row of carpal bones, including Os scaphoideum, Os lunatum and Os triquetrum, should be considered as a functional unit. The arterial supply stems from branches of the rete carpi dorsale et palmare, which pass through the Ligamentum radiocarpeum dorsale et palmare, respectively. In two-thirds of all cases, the scaphoid bone, the most frequently fractured of the wrist bones, is uniformly vascularized, so that by fracture both fragments retain their blood supply. In the remaining third of all cases, only one end of the scaphoid is supplied arterially. Thus the poorly vascularized fragment frequently becomes necrotic.


Loss-of-function of miR-142 by hypermethylation promotes TGF-β-mediated tumour growth and metastasis in hepatocellular carcinoma.
Hypermethylation-induced epigenetic silencing of tumour suppressor genes (TSGs) are frequent events during carcinogenesis. MicroRNA-142 (miR-142) is found to be dysregulated in cancer patients to participate into tumour growth, metastasis and angiogenesis. However, the tumour suppressive role of miR-142 and the status of methylation are not fully understood in hepatocellular carcinoma (HCC). Hepatocellular carcinoma tissues and corresponding non-neoplastic tissues were collected. The expression and function of miR-142 and TGF-β in two HCC cell lines were determined. The miRNA-mRNA network of miR-142 was analysed in HCC cell lines. We found that the miR-142 expression was reduced in tumour tissues and two HCC cell lines HepG2 and SMMC7721, which correlated to higher TNM stage, metastasis and differentiation. Moreover, miR-142 was identified to directly target and inhibit transforming growth factor β (TGF-β), leading to decreased cell vitality, proliferation, EMT and the ability of pro-angiogenesis in TGF-β-dependent manner. Interestingly, the status of methylation of miR-142 was analysed and the results found the hypermethylated miR-142 in tumour patients and cell lines. The treatment of methylation inhibitor 5-Aza could restore the expression of miR-142 to suppress the TGF-β expression, which impaired TGF-β-induced tumour growth. These findings implicated that miR-142 was a tumour suppressor gene in HCC and often hyermethylated to increase TGF-β-induced development of hepatocellular carcinoma.


HspA1A facilitates DNA repair in human bronchial epithelial cells exposed to Benzo[a]pyrene and interacts with casein kinase 2.
Benzo[a]pyrene (BaP) is a ubiquitously distributed environmental pollutant that induces deoxyribonucleic acid (DNA) damage. The inducible heat shock protein (HspA1A) can function as a molecular chaperone; however, its role in DNA repair remains largely unknown. In the present study, human bronchial epithelial cells (16HBE) stably transfected with plasmids carrying HspA1A gene or shRNAs against HspA1A were treated with BaP. DNA damage levels of the cells were evaluated by comet assay. Results suggest that HspA1A could protect cells against DNA damage and facilitate the decrease of DNA damage levels during the first 2 h of DNA repair. DNA repair capacity (DRC) of Benzo(a)pyrene diol epoxide (BPDE)-DNA adducts was evaluated by host cell reactivation assay in the stable 16HBE cells transfected with luciferase reporter vector PCMVluc pretreated with BPDE. Compared with control cells, cells overexpressing HspA1A showed higher DRC (p < 0.01 at 10 μM BPDE and p < 0.05 at 20 μM BPDE, respectively), while knockdown of HspA1A inhibited DNA repair (p < 0.05 at 10 μM BPDE). Moreover, casein kinase 2 (CK2) was shown to interact with HspA1A by mass spectrometry and co-immunoprecipitation assays. The two proteins were co-localized in the cell nucleus and perinuclear region during DNA repair, and were identified by confocal laser scanning microscope. In addition, cells overexpressing HspA1A showed an increased CK2 activity after BaP treatment compared with control cells (p < 0.01). Our results suggest that HspA1A facilitates DNA repair after BaP treatment. HspA1A also interacts with CK2 and enhances the kinase activities of CK2 during DNA repair.