[Acute renal failure in full term neonates with perinatal asphyxia. Prospective study of 87 cases].
Renal involvement is frequent in neonates with perinatal asphyxia. It is correlated with the severity of neurological damage and seems to worsen the long-term neurological outcome. The aim of this study was to determine the incidence of renal failure after perinatal asphyxia, to precise the relationship between severity of cerebral damage and renal failure and to evaluate the place of renal damage in the short- and middle-term neurological outcome. We conducted a prospective study including 87 full-term neonates admitted in the neonatology department of F. Hached university hospital in Sousse (Tunisia) and suffering from hypoxic ischemic encephalopathy from 1st January 2003 to 30 June 2005. Renal function was assessed by measuring plasma urea and creatinine at age 48 h. Renal failure was defined by a level of creatinine above 90 micromol/l. Neurologic examination was performed on day 7. The survivors were followed up by the same senior after discharge. During the study period, 87 full-term neonates were admitted for hypoxic ischemic encephalopathy. The degree of neurological impairment was determined according to Sarnat classification: 1st stage 9 neonates (10,3%), 2nd stage 67 (77%) and 3rd stage 11(12,6%). Renal failure involved 15 neonates (17,2%) of whom 10 belonging to the 2nd stage group. Renal function outcome was favorable in all survivors with normalisation of plasma creatinine level between day 5 and day 15. Eight neonates died, of whom 3 with renal failure. Neurologic examination was abnormal in 36 out of 72 (50%) neonates without renal failure and in 9 of the 12 (75%) survivors with renal failure. Among the 12 neonates with renal failure, 7 had abnormal neurologic features at discharge. Neurologic assessment between 6 and 18 months was abnormal in 4/12 (33%) of neonates with renal failure versus 8/72(11%) of neonates without renal failure. Transient renal failure is commonly observed in perinatal asphyxia. Renal failure is correlated with neurologic severity. Renal function assessment using creatinine plasma level seems to be correlated with neurologic outcome. However, other tools appreciating renal function, namely tubular function, should be determined earlier in order to predict neurologic outcome after hypoxic ischemic encephalopathy.


Neutralization of CD95 ligand promotes regeneration and functional recovery after spinal cord injury.
The clinical outcome of spinal cord injury (SCI) depends in part on the extent of secondary damage, to which apoptosis contributes. The CD95 and tumor necrosis factor (TNF) ligand/receptor systems play an essential role in various apoptotic mechanisms. To determine the involvement of these ligands in SCI-induced damage, we neutralized the activity of CD95 ligand (CD95L) and/or TNF in spinal cord-injured mice. Therapeutic neutralization of CD95L, but not of TNF, significantly decreased apoptotic cell death after SCI. Mice treated with CD95L-specific antibodies were capable of initiating active hind-limb movements several weeks after injury. The improvement in locomotor performance was mirrored by an increase in regenerating fibers and upregulation of growth-associated protein-43 (GAP-43). Thus, neutralization of CD95L promoted axonal regeneration and functional improvement in injured adult animals. This therapeutic strategy may constitute a potent future treatment for human spinal injury.


Flow injection potentiometric determination of total antioxidant activity of plant extracts.
A new flow injection potentiometric (FIP) method, rapid, reproducible and simple to apply, has been developed for the in vitro evaluation of antioxidative capacity of aqueous plant extracts. This method is based on the transient negative signal measurements with a flow-type platinum electrode detector due to the composition change of a [Fe(CN)6]3-/[Fe(CN)6]4- redox-reagent solution. The variables affecting the signal height such as composition and concentration of redox-reagent, injected sample volume, flow rates of carrier and redox-reagent solution streams were studied in details and the conditions were optimized. For the compounds under study, a linear relationship was stated between the potentiometric signal height and the logarithm of antioxidant concentration. It was stated that a wide antioxidant activity range from 1 microM to 10 mM could be determined by the changing concentrations of the hexacyanoferrate(III) from 5 to 0.01 x 10(-4). The present FIP method was applied to quantify relative antioxidant activity (RAA index) of the representative water-soluble antioxidants (ascorbic acid, pyrocatechol, pyrogallol, caffeic acid, chlorogenic acid, gallic acid, tannic acid, uric acid, L-cysteine, trolox). The high sampling rate (100 h(-1)) and a satisfactory reproducibility (R.S.D.=0.7-1.8%, n=5, 0.1 mM each compound) were obtained. The method was also applied to estimate total antioxidant activity (TAA) of real samples (green and black tea infusions, herbal infusions and fresh fruit extracts) and the results were compared with those achieved using well-known in vitro testing methods.


Bladder cancer risk from occupational and environmental exposures.
Approximately 50% of bladder cancer incidence in the United States has been attributed to known carcinogens, mainly from cigarette smoking. Following the identification of this important causative factor, many investigators have attempted to identify other major causes of bladder cancer in the environment. Genetic and epigenetic alterations related to carcinogenesis in the bladder have been linked to environmental and occupational factors unrelated to cigarette smoking and may account for a significant portion of bladder cancer cases in non-smokers. The interaction between genetics and exposures may modulate bladder cancer risk and influence the differing incidence, progression, and mortality of this disease in different genders and races. Comparative molecular studies are underway to measure the relative effects of environment and inheritance to account for observed differences in the epidemiology of bladder cancer. The use of geospatial tools and population-based data will offer further insight into the environmentally-linked causes of bladder cancer.


Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis.
The use of BAL to study ILDs has improved our understanding of IPF pathogenesis. BAL fluid is routinely collected and can be considered a clinical and research tool. The procedure is well tolerated and minimally invasive. No specific cell lines from BAL or immortalized cell lines from IPF patients are available commercially. A method to quickly isolate and characterize fibroblasts from BAL is an unmet research need. Here we describe a new protocol by which we isolated a cell line from IPF. The cell line was expanded in vitro and characterized phenotypically, morphologically and functionally. This culture showed highly filamentous cells with an evident central nucleus. From the phenotypic point of view, this cell line displays fibroblast/myofibroblast-like features including expression of alpha-SMA, vimentin, collagen type-1 and fibronectin. The results showed high expression of ROS in these cells. Oxidative stress invariably promotes extracellular matrix expression in lung diseases directly or through over-production of pro-fibrotic growth factors. Our protocol makes it possible to obtain fibroblasts BAL that is a routine non-invasive method that offers the possibility of having a large sample of patients. Standardized culture methods are important for a reliable model for testing molecules and eventual novel development therapeutic targets.


Evaluation of a novel 8-channel RX coil for speech production MRI at 0.55 T.
Speech production MRI benefits from lower magnetic fields due to reduced off-resonance effects at air-tissue interfaces and from the use of dedicated receiver coils due to higher SNR and parallel imaging capability. Here we present a custom designed upper airway coil for 1H imaging at 0.55 Tesla and evaluate its performance in comparison with a vendor-provided prototype 16-channel head/neck coil. Four adult volunteers were scanned with both custom speech and prototype head-neck coils. We evaluated SNR gains of each of the coils over eleven upper airway volumes-of-interest measured relative to the integrated body coil. We evaluated parallel imaging performance of both coils by computing g-factors for SENSE reconstruction of uniform and variable density Cartesian sampling schemes with R = 2, 3, and 4. The dedicated coil shows approximately 3.5-fold SNR efficiency compared to the head-neck coil. For R = 2 and 3, both uniform and variable density samplings have g-factor values below 1.1 in the upper airway region. For R = 4, g-factor values are higher for both trajectories. The dedicated coil configuration allows for a significant SNR gain over the head-neck coil in the articulators. This, along with favorable g values, makes the coil useful in speech production MRI.


Molecular defects in Sanfilippo syndrome type B (mucopolysaccharidosis IIIB).
Sanfilippo syndrome type B (mucopolysaccharidosis IIIB) is an autosomal recessive disease that is caused by the deficiency of the lysosomal enzyme alpha-N-acetylglucosaminidase (NAGLU). NAGLU is involved in the degradation of the glycosaminoglycan (GAG) heparan sulphate, and a deficiency results in the accumulation of partially degraded GAGs inside lysosomes. Early clinical symptoms include hyperactivity, aggressiveness and delayed development, followed by progressive mental deterioration, although there are a small number of late-onset attenuated cases. The gene for NAGLU has been fully characterized and we report the molecular analysis of 18 Sanfilippo B families. In total, 34 of the 36 mutant alleles were characterized in this study and 20 different mutations were identified including 8 novel changes (R38W, V77G, 407-410del4, 703delT, A246P, Y335C, 1487delT, E639X). The four novel missense mutations were transiently expressed in Chinese hamster ovary cells and all were shown to decrease the NAGLU activity markedly, although A246P did produce 12.7% residual enzyme activity.


Induction of apoptosis in HT-29 colon cancer cells by crude saponin from Platycodi Radix.
This study examined the apoptotic effects of crude saponins acquired from the roots of Platycodon grandiflorum (SPR) in HT-29 human colon cancer cells. SPR decreased HT-29 cell proliferation in dose- and time-dependent manners by inducing apoptosis via DNA fragmentation and poly (ADP-ribose) polymerase (PARP) cleavage. The apoptosis induced by SPR was associated with the activation of initiator caspases-8 and -9, as well as the effector caspase-3. SPR stimulated Bid cleavage, indicating that the apoptotic action of caspase-8-mediated Bid cleavage leads to the activation of caspase-9. SPR increased the expression of the pro-apoptotic protein, Bax, and decreased the expression of the anti-apoptotic protein, Bcl-2. SPR also increased the expression of the caspase-independent mitochondrial apoptosis factor, AIF, in HT-29 cells. These results indicate that SPR inhibits HT-29 cell proliferation by inducing apoptosis, which may be mediated via both caspase-dependent and -independent pathways.


Giant intermuscular lipoma of hip: A case report.
Lipomas are one of the most common benign tumors, but deep tissue and huge lipomas are rare. A 59-year-old middle-aged woman was admitted to hospital for right hip mass resection because of numbness and pain in her right lower limb for 6 months. Ultrasound and nuclear magnetic resonance imaging (MRI) showed irregular mass. On the assumption of malignancy, the patient underwent a right hip mass resection and returned to normal with no short-term or long-term response. HE (hematoxylin-eosin) staining of the tumor showed the characteristics of a benign tumor. The pathogenesis of myolipoma is not clear, and it may be related to brown adipose tissue. When lipoma is huge, it needs interventional treatment. Complete encapsulated myolipoma can usually be completely removed, with low recurrence possibility and good prognosis. Giant intramuscular lipoma of hip and its clinical manifestations are relatively rare. And the results of ultrasound imaging are similar to those of the surrounding normal adipose tissue. No significant difference was observed by naked eye. In particular, postoperative pathological examination is needed to make a definite diagnosis and differential diagnosis. Clinically, surgical treatment is often used, and the prognosis is good, but recurrence is possible. The following report discusses the experience of one of our patients, with literature review.


Heterozygous RELN missense variants associated with genetic generalized epilepsy.
The RELN gene encodes the secreted glycoprotein Reelin and has important functions in both developing and adult brains. In this study, we aimed to explore the association between the RELN and genetic generalized epilepsy (GGE). We performed whole-exome sequencing on a cohort of 92 patients with GGE. Based on amino acid sequence alignments, allele frequency, pedigree validation and computational modeling, the RELN variants were identified and clinical features of cases were summarized. Cell-based Reelin secretion assays were examined by Western blotting. Alterations of mutant Reelin transport through the secretion pathway were detected by immunofluorescence staining. Three novel pathogenic RELN variants (3.26%; c.2260C>T/p.R754W, c.2914C>G/p.P972A and c.3029G>A/p.R1010H) were identified. All probands showed adolescence-onset generalized seizures characterized by generalized epileptiform discharges with normal EEG backgrounds, no or mild cognitive impairment, and responded well to anti-seizure medications. All these variants were located in the central regions from 1B to 2A consecutive repeats, and protein modeling demonstrated structural alterations in Reelin. Moreover, we found that these heterozygous missense variants significantly decreased the secretion of mutant proteins in HEK-293T cells, and this impairment was due to the altered transport of mutant Reelin in the secretion pathway. These results suggest that RELN is potentially associated with GGE. The phenotype of GGE caused by RELN variants is relatively mild, and the pathogenic mechanism may involve a loss-of-function.


Glyceraldehyde-3-phosphate dehydrogenase is a mediator of hemocyte-spreading behavior and molecular target of immunosuppressive factor CrV1.
Cellular immunity is accompanied by hemocyte-spreading behavior, which undergoes cytoskeletal rearrangement. Polydnaviral factor CpBV-CrV1 can inhibit the hemocyte-spreading behavior and suppress host immune response of Plutella xylostella. However, host target molecule of CpBV-CrV1 that inhibits the hemocyte behavior has not been identified yet. This study used a pull-down approach to identify the target molecule of CpBV-CrV1. A protein bound to CpBV-CrV1 was co-precipitated and identified to be glyceraldehyde-3-phosphate dehydrogenase (GAPDH) by LC-MS/MS analysis. RNA interference (RNAi) specific to GAPDH of P. xylostella was found to be able to inhibit the hemocyte-spreading behavior, while RNAi treatments with other glycolytic genes had no effect on the spreading behavior. An addition of recombinant CpBV-CrV1 on hemocyte monolayer interrupted the association between GAPDH and α-tubulin in the cytoplasm. Overlay of mutant proteins (Y492A or Y501A with tyrosine to alanine at putative GAPDH-binding site) of CpBV-CrV1 on hemocyte monolayer revealed that they could enter hemocytes unlike a mutant in the N-terminal coiled-coil domain. However, they failed to inhibit the hemocyte-spreading behavior without any binding affinity to GAPDH. These results suggest that GAPDH plays a critical role in hemocyte-spreading behavior during immune challenge as a molecular target of viral factor CpBV-CrV1.


Antigen-dependent increase in cytosolic free calcium in specific human T-lymphocyte clones.
Calcium has been implicated as an intracellular messenger in the cellular response to various external stimuli. Exposure of lymphocytes to various mitogens and lectins results in rapid transmembrane calcium fluxes and increased cytoplasmic calcium concentrations ([Ca2+]i). It is not clear, however, whether the mechanisms by which these non-physiological stimuli activate cells are related to those involved in antigen-specific activation. We have now used antigen-specific T-cell clones to study changes in [Ca2+]i associated with specific activation and show here that these cells respond specifically in the presence of antigen and antigen-presenting cells (APC) with increased [Ca2+]i and that this increased [Ca2+]i shows the same genetic restrictions as are seen in the proliferation assay. The kinetics of the [Ca2+]i response to antigen indicate that antigen undergoes a time-dependent processing step as a prerequisite for recognition by T cells, as has been shown for T-cell proliferative responses, but that the [Ca2+]i response to processed antigen is extremely rapid. The close correlation between changes in [Ca2+]i and cell activation resulting in proliferation suggests that Ca2+ may act as an intracellular messenger in antigen-specific responses.


Stage of the estrous cycle at the time of pregnant mare's serum gonadotropin injection affects the quality of ovulated oocytes in the mouse.
The present study aims to analyze the effect of the stage of the estrous cycle at the time of pregnant mare's serum gonadotropin (PMSG) injection on number and quality of mouse oocytes retrieved from oviducts after exogenous ovarian stimulation. Cellular and morphological traits of ovulated oocytes from hybrid (C57Bl/6JIco female X CBA/JIco male) female mice of 12, 40-42, 50-52 or 57-62 weeks of age were analyzed. Superovulation was induced by a priming injection of PMSG at different stages of the estrous cycle followed after a 48-hr interval by human chrorionic gonadotropin. Injection of PMSG at diestrus-1 was associated with: (1) increased percentage of cumulus-free oocytes; (2) raised total percentage of oocytes without polar body; (3) increased total percentage of oocytes with intracytoplasmic mitochondrial aggregates; (4) decreased percentage of oocytes with a normal distribution of chromosomes in the metaphase II plate; and (5) raised percentage of oocytes with chromosome scattering when compared to injection at estrus, diestrus-2, and proestrus stage. On the contrary, estrus females displayed the highest percentage of oocytes with a normal distribution of chromosomes in the metaphase II plate and the lowest percentage of oocytes denuded of cumulus cells, without polar body, with intracytoplasmic mitochondrial aggregates and/or with chromosome scattering. These data suggest that administration of gonadotropins in mice should be synchronized with the innate estrous cycle of females to optimize the quality of oocytes collected from oviducts.


High prevalence of hip and groin problems in professional ice hockey players, regardless of playing position.
The prevalence of hip and groin problems in professional male ice hockey is unknown and suspected to differ between playing positions. The purpose of this study was to explore potential differences in the seasonal prevalence of hip and groin problems between playing positions in male elite ice hockey players and to explore the relationship between symptom duration and hip and groin function at the beginning of the new season. Male ice hockey players [n = 329 (92 goalkeepers, 93 defensemen, 144 forwards), Mean age (SD): 24 (5)] from the professional leagues in Sweden responded to an online survey. The survey assessed presence of hip and groin problems (time loss and non-time loss) and symptom duration (categorized into 0, 1-6, or > 6 weeks) in the previous season, and current self-reported hip and groin function (Copenhagen Hip and Groin Outcome Score). During the previous season, 175 players (53.2%) had experienced hip and groin problems. Non time loss problems were experienced by 158 (48%) and time loss problems were experienced by 97 (29.5%) players. No significant differences between playing positions were found. Self-reported function differed significantly between players with different symptom duration and more disability was reported among players with longer symptom duration (p ≤ 0.002). Regardless of playing position, hip and groin problems were prevalent in male ice hockey players. Players with hip and groin problems during the previous season had significantly worse hip and groin function in the beginning of the new season, and longer symptom duration was associated with more disability. III.


The devil is in the details: a synthesis of psychology of facial perception and its applications in forensic facial reconstruction.
The successful identification of a forensic facial reconstruction relies upon many factors other than merely the accuracy of the reconstruction. The way in which the reconstructed head is presented can affect the perception of it, which in turn can affect the recognition rates leading to case resolution. A synthesis of relevant facial perception studies from the psychological literature is provided, followed by a short set of recommendations aimed at practitioners wishing to incorporate the best practice based on current science when presenting their work to the public. These recommendations include pose, background, lighting, and colouring/finishing.


Murine hyperglycemic vasculopathy and cardiomyopathy: whole-genome gene expression analysis predicts cellular targets and regulatory networks influenced by mannose binding lectin.
Hyperglycemia, in the absence of type 1 or 2 diabetes, is an independent risk factor for cardiovascular disease. We have previously demonstrated a central role for mannose binding lectin (MBL)-mediated cardiac dysfunction in acute hyperglycemic mice. In this study, we applied whole-genome microarray data analysis to investigate MBL's role in systematic gene expression changes. The data predict possible intracellular events taking place in multiple cellular compartments such as enhanced insulin signaling pathway sensitivity, promoted mitochondrial respiratory function, improved cellular energy expenditure and protein quality control, improved cytoskeleton structure, and facilitated intracellular trafficking, all of which may contribute to the organismal health of MBL null mice against acute hyperglycemia. Our data show a tight association between gene expression profile and tissue function which might be a very useful tool in predicting cellular targets and regulatory networks connected with in vivo observations, providing clues for further mechanistic studies.


NK cell-mediated targeting of human cancer and possibilities for new means of immunotherapy.
Insights into the molecular basis for natural killer (NK) cell recognition of human cancer have been obtained in recent years. Here, we review current knowledge on the molecular specificity and function of human NK cells. Evidence for NK cell targeting of human tumors is provided and new strategies for NK cell-based immunotherapy against human cancer are discussed. Based on current knowledge, we foresee a development where more cancers may be subject to treatment with drugs or other immunomodulatory agents affecting NK cells, either directly or indirectly. We also envisage a possibility that certain forms of cancers may be subject to treatment with adoptively transferred NK cells, either alone or in combination with other therapeutic interventions.


Interobserver agreement of confocal laser endomicroscopy for detection of head and neck neoplasia.
We have described the feasibility of using the probe-based confocal laser endomicroscopy (pCLE) in differentiating benign from malignant lesions of the head and neck. Therefore, we wanted to determine the interobserver agreement of pCLE offline images of noncancerous, precancerous, and cancerous lesions of the head and neck. Single tertiary referral center. In the feasibility study, image criteria for nondysplasia, dysplasia, and cancer were defined. The pCLE was performed before lesions were biopsied. Fifty offline images and 10 videos of good quality were selected. Seven surgeons and one pathologist were asked to review and categorize the images into the three categories above. The overall accuracy of 29 offline pCLE images and six videos were compared with histopathology. Interobserver agreement and accuracy kappa (κ) scores were measured with 95% confidence intervals (CI). There were six nondysplasia, seven dysplasia, and 11 squamous cell carcinoma (SCCA) cases, each with multiple images. There was substantial agreement between the eight reviewers on the pCLE images and videos (κ = 0.66; 95% CI 0.51-0.82 and κ = 0.71; 95% CI 0.42-0.97, respectively). The overall agreement with the final histopathology was also substantial for both the images and video sequences (κ = 0.70; 95% CI 0.50-0.88 and κ = 0.73; 95% CI 0.39-1.00, respectively). The ability to differentiate normal mucosa, dysplasia, and invasive SCCA using pCLE with high accuracy and reliability was demonstrated. This technology has the potential to decrease sampling error of lesions in the head and neck. This is the first study to test the reliability of this technology in mucosal lesions of the head and neck. NA. Laryngoscope, 126:632-637, 2016.


Hsp and chaperone distribution during endochondral bone development in mouse embryo.
The process of endochondral bone formation was examined with regard to expression of seven heat shock proteins (Hsps): two small Hsps, the constitutive and the inducible forms of the 70 and the 90 Hsp families, the collagen chaperone Hsp47, and a cytosolic chaperone, TCP-1alpha, using immunohistochemistry. Around day 15.5 of embryogenesis the calcification of the long endochondral bones occurs through progressive replacement of the cartilaginous scaffold (rich in type II collagen) with an ossified matrix (rich in type I collagen), and thus a longitudinal section of limb bone recapitulates all the steps of chondrogenesis and the early steps of osteogenesis. We observed that all these Hsps and chaperones are differentially expressed during bone development in a stage-specific pattern reaching very high levels at some specific stages. The involvement of chaperones during these important differentiation steps will be discussed.


The effect of heat on the intestinal and pancreatic levels of amylase and maltase of laying hens and broilers.
The effects of exposing laying hens and broilers daily to intermittent periods of 4 hr heating at 42 degrees C on the intestinal and pancreatic levels of amylase and maltase were investigated. The initial exposure to heat, characterized by heat stress, brought about a significant increase in the duodenum and jejunum parts of the tetra breed hen and only in the duodenum of broilers. The levels of amylase in the distal parts of the intestine of both breeds sharply decreased. The increase in amylase levels in the proximal parts of the intestine under the conditions of initial heating vanished after 3 days of heating; its levels continued to fall in the distal parts. In heat acclimatized laying hens the levels of amylase were lower than those of the control hens both in the intestine and pancreas. The pancreatic level of amylase was reversely related to the levels in the intestine. It is assumed that the intestinal level of amylase is regulated by the pancreas. These findings indicate that the pancreas plays an important role during the adaptation of chickens to heat, through the regulation of intestinal level of amylase. The response in maltase level to heat stress and heat acclimatization was insignificant.


Molecular characterization by high-resolution isoelectric focusing of the products encoded by the class II region loci of the major histocompatibility complex in humans. I. DR and DQ gene variants.
We describe a new approach to the analysis of the structural polymorphism of the DR beta, DQ alpha, and DQ beta polypeptide chains of human histocompatibility class II antigens. In comparison to conventional two-dimensional gel studies, this method provides sharper definition of the protein bands and side-by-side comparisons within the same gel, thereby permitting the detection of minor differences in the isoelectric points of the protein chains. Using this methodology we have analyzed the IEF polymorphism and the variability in the number of the DR beta chains encoded by different DR haplotypes. Twenty DR beta chain variants, which include the products of no less than two separate DR beta loci, have been thus far identified. Alleles at one of these loci are assumed to code for DR beta chains carrying the DR alloespecificities DR1, DR2, DR3, DR4, DR5, DRw6, DR7, and DR8. Alleles at a second DR beta locus encode DR beta chains that may be shared by serologically DR-different haplotypes and carry supertypic serologic specificities (i.e., DRw52 and DRw53). We also demonstrate here that the structural polymorphisms of the DQ alpha and DQ beta chains are more extensive than previously thought, report the characterization of 14 DQ beta variants, and define their relationship to the previously described DQw serologic specificities. In addition, we describe the class II haplotype associations observed for the different DR and DQ variants characterized.


Endoplasmic reticulum stress-related gene model predicts prognosis and guides therapies in lung adenocarcinoma.
The prognosis and survival of lung adenocarcinoma (LUAD) patients are still not promising despite recent breakthroughs in treatment. Endoplasmic reticulum stress (ERS) is a self-protective mechanism resulting from an imbalance in quality control of unfolded proteins when cells are stressed, which plays an active role in lung cancer development, but the relationship between ERS and the pathological characteristics and clinical prognosis of LUAD patients remains unclear. LASSO and Cox regression were applied based on sequencing information to construct the model, which was validated to be robust. The risk scores of the patients were calculated using the formula provided by the model, and the patients were divided into high and low-risk groups according to the median cut-off of risk scores. Cox regression analysis identifies independent prognostic factors for these patients, and enrichment analysis of prognosis-related genes was also performed. The relationship between risk scores and tumor mutation burden (TMB), cancer stem cell index, and drug sensitivity was explored. We constructed a 13-gene prognostic model for LUAD patients. Patients in the high-risk group had worse overall survival, lower immune score and ESTIMATE score, higher TMB, higher cancer stem cell index, and higher sensitivity to conventional chemotherapeutic agents. In addition, we constructed a nomogram that predicts 5-year survival in LUAD patients, which helps clinicians to foresee the prognosis from a new perspective. Our results highlight the association of ERS with LUAD and the potential use of ERS in guiding treatment.


Evaluation of the association of chronic inflammation and cancer: Insights and implications.
Among the most extensively researched processes in the development and treatment of cancer is inflammatory condition. Although acute inflammation is essential for the wound healing and reconstruction of tissues that have been damaged, chronic inflammation may contribute to the onset and growth of a number of diseases, including cancer. By disrupting the signaling processes of cells, which result in cancer induction, invasion, and development, a variety of inflammatory molecules are linked to the development of cancer. The microenvironment surrounding the tumor is greatly influenced by inflammatory cells and their subsequent secretions, which also contribute significantly to the tumor's growth, survivability, and potential migration. These inflammatory variables have been mentioned in several publications as prospective diagnostic tools for anticipating the onset of cancer. Targeting inflammation with various therapies can reduce the inflammatory response and potentially limit or block the proliferation of cancer cells. The scientific medical literature from the past three decades has been studied to determine how inflammatory chemicals and cell signaling pathways related to cancer invasion and metastasis are related. The current narrative review updates the relevant literature while highlighting the specifics of inflammatory signaling pathways in cancer and their possible therapeutic possibilities.


Prevalence and Trajectory of COVID-19-Associated Hypercoagulability Using Serial Thromboelastography in a South African Population.
The coagulation abnormalities resulting from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been attributed to inflammation and subsequent cytokine storm. Thromboelastography (TEG) is a point-of-care test used to assess clot formation and degradation in whole blood and is an indicator of the overall real-time coagulopathic state of the patient. A single-centre, prospective, observational cohort study was conducted in South Africa, analysing the coagulation patterns of 41 patients with hypoxia related to SARS-CoV-2 using serial thromboelastography (TEG) on admission, after 48 hours, and at resolution of hypoxia/day 10. Results: Two-thirds (n = 26) were women. The median age was 61 (IQR 50-67), and the majority (88%) were Black patients. Almost half (22) of the patients were critically ill and ventilated, with median SOFA and SAPS2 scores of 3 and 22 (IQR2-4 and 18-30), respectively. The prevalence of hypercoagulability was 0.54 (95% CI 0.46-0.62), whilst 29/41 (0.71, CI 0.64-0.78)) met the definition of hypofibrinolysis. Differences between the hypercoagulable (HC) and non-hypercoagulable groups remained apparent at 48 hours after anticoagulation. At this time point, the K time was significantly lower (p ˂ 0,01), and the α-angle (p ˂ 0,01) and maximum amplitude (MA) (p ˂ 0,01) were significantly higher in the HC cohort. At resolution of hypoxia, or day 10, only MA was significantly higher in the hypercoagulable group compared to the non-hypercoagulable group (p = 0.01). The initial impairment in fibrinolysis (Ly30), α angle, and MA were significantly associated with mortality, with p values of 0.006, 0.031, and 0.04, respectively. In this South African population, hypercoagulability was a highly prevalent phenomenon in COVID-19 disease. It was typified by hypofibrinolysis and a persistently elevated MA, despite anticoagulation therapy.


Presenting complaints and findings in a group of patients attending a hypodontia clinic.
A survey is presented of the complaints and findings in a group of 451 patients referred to a multidisciplinary clinic for the management of hypodontia. The most common complaints were missing teeth, spacing in the dental arches, and poor appearance. The majority of patients were referred by their dental practitioners, and more than half were over 12 years old when first seen. Delayed referral can have educational and social consequences. Changing attitudes to dental care and the maintenance of an intact dentition are resulting in hypodontia, particularly in its more severe forms, becoming of increasing concern to children and their parents. The management of this condition is best organised via multidisciplinary specialist clinics.


Evaluation of indoor residual spraying with the pyrrole insecticide chlorfenapyr against pyrethroid-susceptible Anopheles arabiensis and pyrethroid-resistant Culex quinquefasciatus mosquitoes.
Chlorfenapyr is a pyrrole insecticide with a unique non-neurological mode of action. Laboratory bioassays of chlorfenapyr comparing the mortality of pyrethroid-susceptible and -resistant Anopheles gambiae s.s. and Culex quinquefasciatus mosquitoes indicated that operational cross-resistance is unlikely to occur (resistance ratio ranged between 0 and 2.1). Three trials of chlorfenapyr indoor residual spraying were undertaken in experimental huts in an area of rice irrigation in northern Tanzania that supports breeding of A. arabiensis. Daily mosquito collections were undertaken to assess product performance primarily in terms of mortality. In the second trial, 250mg/m(2) and 500mg/m(2) chlorfenapyr were tested for residual efficacy over 6 months. Both dosages killed 54% of C. quinquefasciatus, whilst for A. arabiensis 250mg/m(2) killed 48% compared with 41% for 500mg/m(2); mortality was as high at the end of the trial as at the beginning. In the third trial, 250mg/m(2) chlorfenapyr was compared with the pyrethroid alpha-cypermethrin dosed at 30mg/m(2). Chlorfenapyr performance was equivalent to the pyrethroid against A. arabiensis, with both insecticides killing 50% of mosquitoes. Chlorfenapyr killed a significantly higher proportion of pyrethroid-resistant C. quinquefasciatus (56%) compared with alpha-cypermethrin (17%). Chlorfenapyr has the potential to be an important addition to the limited arsenal of public health insecticides for indoor residual control of A. arabiensis and pyrethroid-resistant species of mosquito.


Oxidative stress, DNA damage, and gene expression induced by flufiprole enantiomers in the earthworms (Eisenia fetida).
To evaluate flufiprole enantiomers ecotoxicology and early warning to earthworms in the soil, the effect of flufiprole enantiomers on the activity of antioxidative enzymes (superoxide dismutase, SOD; catalase, CAT; peroxidase, POD), malondialdehyde (MDA) content, DNA damage and expression level of target genes (SOD, TCTP, l-rRNA, and HSP90) have been investigated. Compared to the controls, the activities of SOD, CAT, POD, and MDA content were first inhibited and then stimulated by all concentrations of flufiprole enantiomers in 28 days. The olive tail moment (OTM) value was maximum on the 14th day, then decreased gradually, and the higher the concentration, the slower the decrease. The expression level of the SOD and TCTP genes increased first and then decreased. In conclusion, the toxicity of flufiprole enantiomers to earthworms is: R-(+)- flufiprole >Rac-(±)- flufiprole >S-(-)- flufiprole. The SOD could be the key biomarker for monitoring the risk of flufiprole enantiomers.


Experimental investigation on mechanical properties of aged bamboo fiber-reinforced composites under quasi-static loading.
Bamboo fiber-reinforced composites (BFRC) have gradually attracted the interest of researchers due to their excellent mechanical properties. However, the age factor can affect these composites' mechanical properties and weaken them. This study aims to explore the mechanical properties of BFRC subjected different aging tests. First, the BFRC is prepared from raw bamboo through a series of technological processes. Second, the BFRC sheets are processed into tensile and compressive samples with different directions. Third, the samples are treated with a cold-hot test and a UV dry-wet test, respectively. Finally, quasi-static experiments are carried out to study the mechanical properties of aged BFRC. The results reveal that the BFRC has significant anisotropy. The tensile strength of BFRC parallel with the axis of the reinforcing fiber is as high as 148.53 MPa. It is 26.47 times greater than the tensile strength of BFRC perpendicular to the axis of the reinforcing fiber. The mechanical responses and SEM images show that the UV dry-wet test significantly affects on the BFRC compared to the cold-hot test. It dramatically reduces the mechanical properties of the BFRC. The SEM images further indicate that cracks occur on the surface of BFRC samples under the UV dry-wet test. In addition, two aging tests have an interactive effect on BFRC, which can reduce the sensitivity of BFRC to a UV dry-wet environment. These results are expected to deepen the understanding of the mechanical properties of BFRC and provide guidance for its applicable conditions.


Automated quantification of pulmonary perfused blood volume by dual-energy CTPA in chronic thromboembolic pulmonary hypertension.
The aim of the study was to determine whether automated quantification of pulmonary perfused blood volume (PBV) in dual-energy computed tomography pulmonary angiography (DE-CTPA) can be used to assess the severity of chronic thromboembolic pulmonary hypertension (CTEPH). Automated quantification of PBV was performed in 25 consecutive CTEPH patients undergoing DE-CTPA. PBV values were correlated with cardiac index and pulmonary vascular resistance quantified by right heart catheterization and walking distance in the 6-minute walk test using Pearson's correlation coefficient and multivariate linear regression analysis to control for age and gender. DE-CTPA derived PBV values inversely correlated with systolic (r = -0.64, p = 0.001) and mean (r = -0.57, p = 0.004) pulmonary arterial pressure. There was a trend for PBV values to inversely correlate with pulmonary vascular resistance (r = -0.20, p = 0.35). No significant correlation was found between PBV values and cardiac index or 6-minute walking distance. These correlations were confirmed to be independent of age and gender on multivariate linear regression analysis. DE-CTPA can be used for an automated quantification of pulmonary PBV in chronic thromboembolic pulmonary hypertension. PBV values correlate inversely with systolic and mean pulmonary arterial pressure and can thus be used to estimate the severity of pulmonary hypertension in these patients. Citation Format: • Meinel FG, Graef A, Thierfelder KM et al. Automated Quantification of Pulmonary Perfused Blood Volume by Dual-Energy CTPA in Chronic Thromboembolic Pulmonary Hypertension. Fortschr Röntgenstr 2014; 186: 151 - 156.


Transparent nanocellulose paper-based biodegradable colorimetric nerve agent detectors.
Transparent cellulose nanofiber (t-CNF) films were prepared by succinylation and an aqueous collision counter system treatment, and used for the colorimetric detection of diethyl chlorophosphate (DCP), a nerve agent mimic in the vapor phase. DCP receptor with an oxime residue was anchored on the surface of succinylated CNF films, resulting in the target probe (CNF-Azo films). CNF-Azo films exhibited selective detection behavior toward DCP in the vapor phase. The oxime groups of CNF-Azo film reacted with DCP upon exposure to DCP vapor, which was accompanied by a color change from yellow to purple. Significantly, the film's transparency was preserved throughout the detection process, allowing it to identify objects behind the film during DCP detection. This property could apply to any detection system in which the color change caused by detection does not interfere with the film's transparency. The CNF-based film sensor was biodegradable, allowing it to be disposable after use.


A decision analysis study of the value of a liver biopsy in nonalcoholic steatohepatitis.
Nonalcoholic fatty liver disease (NAFLD) is an increasingly prevalent clinical syndrome reflecting a mixture of pathological liver histology with the potential to progress to cryptogenic cirrhosis. Liver biopsy (LB) is the only way to determine the severity of disease. However, LB is invasive, has complications and financial costs. The aim of this study is to determine the benefits of early LB for diagnosis and early treatment. A decision tree model tracked the potential outcomes of NAFLD between an LB-directed approach vs. no initial LB. Each specific biopsy category determined subsequent care actions. The baseline probabilities were determined by literature review and expert opinion. An initial LB strategy was projected to have a lower mortality compared with the no initial LB group and fewer transplant eligible patients after 5 years. If the prevalence of NAFLD is increased, the mortality increases in both the study groups. With improved efficacy of medical and/or surgical intervention, the mortality rate was decreased. This decision tree model provides a context for balancing the risk and benefit of LB in NAFLD. With early biopsy, and early intervention, the relative return of preventing advanced liver disease per LB was high.


Matrix metalloproteinase-9 regulates the blood brain barrier via the hedgehog pathway in a rat model of traumatic brain injury.
The mechanisms of secondary brain injury after traumatic brain injury (TBI) are complex and are the result of multiple factors. Protecting the blood-brain barrier (BBB) and ameliorating cerebral edema are two key factors for improving the prognosis of TBI patients. The BBB is regulated by the hedgehog pathway through Scube2 and Shh protein. Matrix metalloproteinase-9 (MMP-9) influences the transport system and enzyme system of vascular endothelial cells, possibly via the hedgehog pathway. The present study aimed to investigate the role and mechanism of MMP-9 in TBI via the hedgehog pathway. Eighty male Sprague-Dawley rats were used to establish a murine model of TBI. Subsequently, the effect of SB-3CT-a specific inhibitor of MMP-9-was assessed via Western blotting, real-time PCR, immunofluorescence, apoptotic assays, and neurological scoring. The results showed that, compared with those of the sham-operation group, the mRNA and protein levels of MMP-9 were significantly increased after TBI, while the expressions of Scube2 and Shh were decreased. Application of SB-3CT at 24 h after TBI significantly reduced neuronal apoptosis and BBB permeability, while increasing expressions of Scube2 and Shh. In conclusion, these findings demonstrate an influence of TBI-induced MMP-9 upregulation in the induction of post-traumatic nerve and BBB injury, which may be partially mediated by Scube2 and Shh via the hedgehog pathway.


Influence of donor and recipient sex on outcomes following simultaneous pancreas-kidney transplantation in the new millennium: Single-center experience and review of the literature.
The influence of sex on outcomes following simultaneous pancreas-kidney transplantation (SPKT) in the modern era is uncertain. We retrospectively studied 255 patients undergoing SPKT from 11/2001 to 8/2020. Cases were stratified according to donor (D) sex, recipient (R) sex, 4 D/R sex categories, and D/R sex-matched versus mismatched. D-male was associated with slightly higher patient (p = .08) and kidney (p = .002) but not pancreas (p = .23) graft survival rates (GSR) compared to D-female. There were no differences in recipient outcomes other than slightly higher pancreas thrombosis (8% R-female vs. 4.2% R-male, p = .28) and early relaparotomy rates in female recipients (38% R-female vs. 29% R-male, p = .14). When analyzing the 4 D/R sex categories, the two D-male groups had higher kidney GSRs compared to the two D-female groups (p = .01) whereas early relaparotomy and pancreas thrombosis rates were numerically higher in the D-female/R-female group compared to the other three groups. Finally, there were no significant differences in outcomes between sex-matched and sex-mismatched groups although overall survival outcomes were lower with female donors irrespective of recipient sex. The influence of D/R sex following SPKT is subject to multiple confounding issues but survival rates appear to be higher in D-male/R-male and lower in D-female/R-male categories.


Pharmacokinetics and toxicity profiling of 4-(methylthio)butyl isothiocyanate with special reference to pre-clinical safety assessment studies.
4-(methylthio)butyl isothiocyanate (4-MTBITC) also called erucin is abundantly present in the seeds of Eruca sativa plant closely related to cruciferous vegetables rich in isothiocyanates. We have previously reported the molecular targets of 4-MTBITC, but no acute, subacute and subchronic toxicity studies have been carried out to evaluate its safety. The non-everted gut sac method was used to study intestinal absorption and it revealed the highest absorption of 4-MTBITC in the jejunum. Dose-dependent pharmacokinetic parameters were observed in rats given 10, 20, and 40 mg/kg oral doses of 4-MTBITC. At the highest dose of 40 mg/kg, Cmax was 437.33 μg/ml and Tmax was 30 min, suggesting quick absorption and delayed elimination with elimination constant, 0.0036 ± 0.0002min-1. In a 14 days toxicity study, the mean LD50 of 4-MTBITC was 500 mg/kg body weight. After 28 and 90 days of treatment with 4-MTBITC (2.5, 10, 40 mg/kg/day), significant increases were observed in SGOT, cholesterol, and antioxidant enzymes. The levels of glycine, alanine and lysine were markedly increased in the liver tissue, thereby indicating that the liver was the target organ of 4-MTBITC induced toxicity in female animals. The histopathological examination of liver, kidney, and lung tissues revealed little focal necrosis, apoptosis, and reduction in the levels of amino acids involved in cellular metabolic pathways, indicating the anti-proliferative potential of 4-MTBITC against rapidly growing cells.


Monitoring heavy metals in the Gulf of Thailand using mussel watch approach.
Concentration of zinc, manganese, copper, chromium, nickel and cadmium were determined in the whole soft parts of the mussel, Perna viridis, collected along the coast of the Gulf of Thailand. Mussels were sampled from 10 locations, during 1989-1990, following an earlier study in 1986. The trends of accumulations of investigated metals in the mussel were not higher than the previous study, except for the concentration of manganese and nickel. The levels of manganese and nickel in the mussel in 1989 and in 1990 were much higher than those reported previously for 1986. In addition, the level of cadmium, manganese and chromium in mussels from the western coast of the Gulf was higher than that from the other parts of the Gulf. However, the levels of such metals were within the acceptable limits with respect to the public health standards.


Heritable choice of colony size in cliff swallows: does experience trump genetics in older birds?
The variation in breeding-colony size seen in populations of most colonial birds may reflect heritable choices made by individuals who are phenotypically specialized for particular social environments. Although a few studies have reported evidence for genetically based choice of group sizes in birds, we know relatively little about the extent to which animals potentially rely on experience versus innate preferences in deciding with how many conspecifics to settle at different times of their lives. We conducted a cross-fostering experiment in 1997-1998 on cliff swallows (Petrochelidon pyrrhonota) in southwestern Nebraska, USA, in which some individuals were reared in colonies different in size from those in which they were born. Breeding-colony sizes chosen by this cohort of birds were monitored by mark-recapture throughout their lives. A multistate mark-recapture analysis revealed that birds in their first breeding year chose colony sizes similar to those of their birth, regardless of their rearing environment, confirming a previous analysis. Beyond the first breeding year, however, cliff swallows' colony choice was less dependent on where they were born. Birds born in small colonies and reared in large colonies showed evidence of a delayed rearing effect, with these birds overwhelmingly choosing large colonies in later years. Heritabilities suggested strong genetic effects on first-year colony choice but not in later years. Cliff swallows' genetically based colony-size preferences their first year could be a way to ensure matching of their phenotype to an appropriate social environment as yearlings. In later years, familiarity with particular colony sites and available information on site quality may override innate group-size preferences when birds choose colonies.


High-grade B-cell lymphoma: a term re-purposed in the revised WHO classification.
The designation high-grade B-cell lymphoma traditionally has been used as a general term for B-cell lymphomas that are morphologically aggressive with many mitotic figures, often a starry-sky pattern, and a high proliferation rate as shown by Ki-67 expression. These morphological features correlate with aggressive clinical behaviour. In the 2017 revision of the World Health Organization (WHO) classification of lymphomas, the term high-grade B-cell lymphoma has been re-purposed. Most cases in this category include so-called double and triple-hit lymphomas in the subgroup designated high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements. A smaller subset of cases are designated as high-grade B-cell lymphoma, not otherwise specified (NOS). This group is stated to be heterogeneous and includes neoplasms without double/triple-hit genetics as well as neoplasms that do not readily fit within the categories of diffuse large B-cell lymphoma NOS or Burkitt lymphoma. In most cases, accurate diagnosis can be achieved by integration of morphological, immunophenotypic and conventional cytogenetic and/or fluorescence in situ hybridisation (FISH) results. However, a few cases of high-grade B-cell lymphoma show overlapping features with different entities and can cause diagnostic difficulties. The WHO scheme is generally based on a 'snapshot' of current knowledge and, in our opinion, does not entirely capture the current state of knowledge of high-grade B-cell lymphomas. In this article, we review the topic of high-grade B-cell lymphoma as is currently described in the WHO classification, address recent developments, discuss difficulties one can encounter when applying the WHO scheme in daily practice, and we present some suggestions for the diagnostic workup of high-grade B-cell lymphomas.


Programmed death ligand-1 expression and occult lymph node metastasis in non-small cell lung cancer.
Identifying the preoperative risk factors for lymph node upstaging could contribute to the development of individualized perioperative treatment for patients with non-small cell lung cancer (NSCLC). The current study aimed to evaluate the risk factors for lymph node upstaging, including gene mutation and programmed death ligand-1 expression in patients with resectable NSCLC. Data on the clinicopathological characteristics of patients who underwent lobectomy for clinical N0 NSCLC at our institution were collected. The clinicopathological findings of the pathological N0 and lymph node upstaging groups were then analyzed. Univariate and multivariate analyses were performed to examine the predictive factors for nodal upstaging. Of 291 patients, 40 had postoperative nodal upstaging (n = 25, N1; n = 15, N2). Large tumor size and high maximum standardized uptake value were significantly associated with nodal upstaging. The nodal upstaging group had a higher proportion of patients with solid adenocarcinoma and lymphatic, vascular, and pleural invasion than the pathological N0 group. Further, the nodal upstaging group had a higher proportion of patients with positive programmed death ligand-1 expression than the pathological N0 group. Univariate and multivariate analyses showed that tumor size and positive programmed death ligand-1 expression were associated with nodal upstaging. The appropriate therapeutic strategy including preoperative treatment and resection should be cautiously considered preoperatively in patients with clinical N0 NSCLC who have large tumors and positive programmed death ligand-1 expression.


Deuterium-labelled N-acyl-L-homoserine lactones (AHLs)--inter-kingdom signalling molecules--synthesis, structural studies, and interactions with model lipid membranes.
N-Acyl-L-homoserine lactones (AHLs) are synthesized by Gram-negative bacteria. These quorum-sensing molecules play an important role in the context of bacterial infection and biofilm formation. They also allow communication between microorganisms and eukaryotic cells (inter-kingdom signalling). However, very little is known about the entire mechanism of those interactions. Precise structural studies are required to analyse the different AHL isomers as only one form is biologically most active. Theoretical studies combined with experimental infrared and Raman spectroscopic data are therefore undertaken to characterise the obtained compounds. To mimic interactions between AHL and cell membranes, we studied the insertion of AHL in supported lipid bilayers, using vibrational sum-frequency-generation spectroscopy. Deuterium-labelled AHLs were thus synthesized. Starting from readily available deuterated fatty acids, a two-step procedure towards deuterated N-acyl-L-homoserine lactones with varying chain lengths is described. This included the acylation of Meldrum's acid followed by amidation. Additionally, the detailed analytical evaluation of the products is presented herein.


Cytoplasmic p27Kip1 promotes tumorigenesis via suppression of RhoB activity.
The cell cycle inhibitor p27Kip1 is a tumor suppressor via the inhibition of CDK complexes in the nucleus. However, p27 also plays other functions in the cell and may acquire oncogenic roles when located in the cytoplasm. Activation of oncogenic pathways such as Ras or PI3K/AKT causes the relocalization of p27 in the cytoplasm, where it can promote tumorigenesis by unclear mechanisms. Here, we investigated how cytoplasmic p27 participates in the development of non-small cell lung carcinomas. We provide molecular and genetic evidence that the oncogenic role of p27 is mediated, at least in part, by binding to and inhibiting the GTPase RhoB, which normally acts as a tumor suppressor in the lung. Genetically modified mice revealed that RhoB expression is preferentially lost in tumors in which p27 is absent and maintained in tumors expressing wild-type p27 or p27CK- , a mutant that cannot inhibit CDKs. Moreover, although the absence of RhoB promoted tumorigenesis in p27-/- animals, it had no effect in p27CK- knock-in mice, suggesting that cytoplasmic p27 may act as an oncogene, at least in part, by inhibiting the activity of RhoB. Finally, in a cohort of lung cancer patients, we identified a subset of tumors harboring cytoplasmic p27 in which RhoB expression is maintained and these characteristics were strongly associated with decreased patient survival. Thus, monitoring p27 localization and RhoB levels in non-small cell lung carcinoma patients appears to be a powerful prognostic marker for these tumors. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


Physical and transcript map of the region between D6S264 and D6S149 on chromosome 6q27, the minimal region of allele loss in sporadic epithelial ovarian cancer.
We have previously shown a high frequency of allele loss at D6S193 (62%) on chromosomal arm 6q27 in ovarian tumours and mapped the minimal region of allele loss between D6S297 and D6S264 (3 cM). We isolated and mapped a single non-chimaeric YAC (17IA12, 260-280 kb) containing D6S193 and D6S297. A further extended bacterial contig (between D6S264 and D6S149) has been established using PACs and BACs and a transcript map has been established. We have mapped six new markers to the YAC; three of them are ESTs (WI-15078, WI-8751, and TCP10). We have isolated three cDNA clones of EST WI-15078 and one clone contains a complete open reading frame. The sequence shows homology to a new member of the ribonuclease family. The other two clones are splice variants of this new gene. The gene is expressed ubiquitously in normal tissues. It is expressed in 4/8 ovarian cancer cell lines by Northern analysis. The gene encodes for a 40 kDa protein. Direct sequencing of the gene in all the eight ovarian cancer cell lines did not identify any mutations. Clonogenic assays were performed by transfecting the full-length gene in to ovarian cancer cell lines and no suppression of growth was observed.


Severe uvular edema and nasal mucosal necrosis due to Ecbalium elaterium (squirting cucumber): an allergic reaction or direct toxic effect?
Ecbalium elaterium or squirting cucumber is a plant growing in the Mediterranean region, belong to Cucurbitaceae family. Its juice has been used as an herbal medicine since ancient times because of its cathartic and anti-inflammatory effects. It has been reported to cause upper airway edema, especially after the administration of its undiluted form. It has generally been attributed to allergic reaction. In this case report we aimed to interrogate this anticipation by presenting a case of severe uvular edema and nasal mucosal necrosis after dripping undiluted Ecbalium elaterium juice that did not improved by anti-allergic therapies.


Results of topography-guided laser in situ keratomileusis custom ablation treatment with a refractive excimer laser.
To evaluate the safety and effectiveness of topography-guided custom ablation treatment (T-CAT) to correct myopia and myopic astigmatism with laser in situ keratomileusis (LASIK). Nine clinical sites in the USA. Prospective observational nonrandomized unmasked study. The study comprised patients aged 18 to 65 years old with myopia or myopic astigmatism with a manifest refraction spherical equivalent (MRSE) up to -9.0 diopters (D) and astigmatism of 6.0 D or less. Patients with previous refractive surgery or abnormal topography were excluded. Corneal topographies were obtained using the Allegro Topolyzer, and laser treatment was delivered with the Allegretto Wave Eye-Q excimer laser system. Visual outcomes were evaluated postoperatively at 1 day, 1 week, and 1, 3, 6, 9, and 12 months. The clinical trial enrolled 212 patients (249 eyes). The T-CAT procedure significantly reduced the MRSE and cylinder, with stability of outcomes evident from 3 to 12 months after surgery. Compared with the preoperative corrected distance visual acuity (CDVA), the postoperative uncorrected distance visual acuity (UDVA) improved by 1 line or more in 30% of eyes and the postoperative UDVA was at least as good as the preoperative CDVA in 90% of eyes. Most visual symptoms improved after T-CAT. There were no significant treatment-related adverse events or loss of vision. The T-CAT procedure performed with the diagnostic device and the refractive excimer laser system safely and effectively achieved predictable refractive outcomes and reduced visual symptoms with stable results through 12 months. Dr. Stulting is a paid consultant to Alcon Laboratories, Inc., and was a medical monitor for the U.S. Food and Drug Administration (FDA) clinical trial. Dr. Fant is president of Clinical Research Consultants, Inc. (CRC), the clinical and regulatory consulting group that sponsored the FDA clinical trial. Dr. Fant and CRC were supported by Alcon Laboratories, Inc.


Prosthetic treatment for chemical burns of the oral cavity.
Chemical burns of the mouth, pharynx, and digestive passages have been reviewed in regard to the nature of the injury and wound description. Fabrication of custom mouth protectors to minimize oral scar contracture and its sequelae has been described. Mouth protectors provide flexible but active resistance to the forces of scar contracture and avoid a constant unyielding force that could cause pressure necrosis. Protectors are easy to fabricate and adjust, and permit movement and function of the injured area.


The new occlutech duct occluder: immediate results, procedural challenges, and short-term follow-up.
The aim of this study was to assess the immediate and short-term results of transcatheter closure of patent ductus arteriosus (PDA) using the Occlutech duct occluder (ODO). The ODO is a new self-expanding nitinol device. Its body is wider at the pulmonary artery end than the aortic end. It is available in longer lengths than the Amplatzer duct occluder. Twenty-two ODO implants were attempted in successive children referred for transcatheter device occlusion of PDAs ≥1 mm in diameter. Median patient age was 2.4 years (range, 0.7-17.5 years), median weight was 13.1 kg (range, 6.3-40 kg), and median PDA diameter was 1.9 mm (range, 1-4.3 mm). Twenty-one out of 22 patients (95%) had successful ODO implantation. One device was withdrawn before release because it did not reach the pulmonary artery end of a long duct. Median procedure time was 40 minutes (range, 26-60 minutes) and fluoroscopy time was 4.5 minutes (range, 2.7-13.3 minutes). Occlusion rates were 19/21 (90%) at the end of the procedure, 20/21 (95%) at 24-48 hours post procedure, and 21/21 (100%) on echocardiography at a median follow-up of 4 weeks (range, 2-16 weeks). There were no device-related complications. Two infants had femoral artery occlusion, successfully treated by heparinization and thrombolysis. This first formal clinical evaluation of the ODO indicates that it is safe and effective in occluding small-to-moderate size ducts, up to a diameter of 4.3 mm. The device produced equivalent results to the Amplatzer duct occluder. Further evaluation is required to assess whether its shape and longer length make it superior for closing large and long ducts.


Expression of novel β-glucanase Cel12A from Stachybotrys atra in bacterial and fungal hosts.
β-glucanase Cel12A from Stachybotrys atra has been cloned and expressed in Aspergillus niger. The purified enzyme showed high activity of β-1,3-1,4-mixed glucans, was also active on carboxymethylcellulose (CMC), while it did not hydrolyze crystalline cellulose or β-1,3 glucans as laminarin. Cel12A showed a marked substrate preference for β-1,3-1,4 glucans, showing maximum activity on barley β-glucans (27.69 U mg(-1)) while the activity on CMC was much lower (0.51 U mg(-1)). Analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focussing (IEF), and zymography showed the recombinant enzyme has apparent molecular weight of 24 kDa and a pI of 8.2. Optimal temperature and pH for enzyme activity were 50°C and pH 6.5. Thin layer chromatography analysis showed that major hydrolysis products from barley β-glucan and lichean were 3-O-β-cellotriosyl-D-glucose and 3-O-β-cellobiosyl-D-glucose, while glucose and cellobiose were released in smaller amounts. The amino acid sequence deduced from cel12A revealed that it is a single domain enzyme belonging to the GH12 family, a family that contains several endoglucanases with substrate preference for β-1,3-1,4 glucans. We believe that S. atra Cel12A should be considered as a lichenase-like or nontypical endoglucanase.


Regulation of adrenal atrial natriuretic hormone receptor subtypes.
Regulation of atrial natriuretic hormone (ANH) receptor binding and aldosterone suppression was studied in isolated adrenal glomerulosa cells from rats fed a high-salt (HS) or low-salt (LS) diet for 3 days. In plasma of HS rats, aldosterone levels were 5 times lower and immunoreactive ANH two times higher than in LS rats. Competitive binding studies showed the same affinity for human atrial natriuretic hormone (hANH) in both pools of cells, but receptor density was 50% higher on LS cells. A linear ANH analog that binds to non-guanylate-cyclase-coupled receptors did not show increased binding to LS cells. Cyclic GMP production in response to hANH was identical in both groups. The aldosterone-inhibitory effect of hANH on both groups of basal and angiotensin II-stimulated cells was also identical. Thus a short-term high-salt diet causes decreased density of ANH receptors in glomerulosa cells without changing biological activity of ANH. These results suggest that dietary salt content changes the number of ANH receptors and that non-guanylate-cyclase-coupled receptors contain at least two classes of receptors.


Two-Step 3D-Guided Supramalleolar Osteotomy to Treat Varus Ankle osteoarthritis.
Success of valgus-type supramalleolar osteotomy (SMOT) depends on adequate correction of malalignment, which can be hard to achieve with current 2-dimensional (2D) planning and operative techniques. A personalized digital 3-dimensional (3D) workflow to virtually plan and perform a 2-step 3D-guided medial opening (MO) SMOT has the potential to improve precision of correction. Computed tomography (CT)-based Proplan medical 3D models were made to virtually plan the desired MO SMOT, and exported to 3-Matic medical to develop patient-specific 2-step cutting and wedge guides. Workflow accuracy was tested in this limited clinical pilot study (3 patients) by comparing the virtual planned position of the osteotomized distal tibial fragment with the 1-year post-MO SMOT configuration. Two millimeters or less translation deviation in every plane was defined as accurate. Primary outcome analysis of the osteotomized distal tibial fragment deviation showed a median translation in all planes of 0.7 (range 0-8.2) mm (interquartile range 1.55) with an excellent interrater reliability of the measurements (intraclass correlation coefficient 0.998). There was a strong reduction in ankle pain as reflected by an increase of the AOFAS-AH score and decrease of NRS pain score with an unrestricted hindfoot motion 1 year after surgery. 3D virtually planned bone cutting and wedge guides is a promising approach associated with minimal postoperative deviation from the desired correction in medial opening supramalleolar osteotomy.


Enterococcus faecalis bloodstream infection: does infectious disease specialist consultation make a difference?
To evaluate the relationship between mortality or relapse of bloodstream infection (BSI) due to Enterococcus faecalis and infectious diseases specialist consultation (IDC) and other factors potentially associated with outcomes. In a tertiary-care center, consecutive adult patients with E. faecalis BSI between January 1, 2016 and January 31, 2019, were prospectively followed. The management of E. faecalis BSI was evaluated in terms of adherence to evidence-based quality-of-care indicators (QCIs). IDC and other factors potentially associated with 90-day-mortality or relapse of E. faecalis BSI were analyzed by multivariate logistic regression. A total of 151 patients with a median age of 68 years were studied. IDC was performed in 38% of patients with E. faecalis BSI. 30 cases of endocarditis (20%) were diagnosed. All-cause in-hospital mortality was 23%, 90-day mortality was 37%, and 90-day relapsing E. faecalis BSI was 8%. IDC was significantly associated with better adherence to 5 QCIs. Factors significantly associated with 90-day mortality or relapsing EfB in multivariate analysis were severe sepsis or septic shock at onset (HR 4.32, CI 2.36e7.88) and deep-seated focus of infection (superficial focus HR 0.33, CI 0.14e0.76). Enterococcus faecalis bacteremia is associated with a high mortality. IDC contributed to improved diagnostic and therapeutic management.


Winnie the Pooh and psoriasis too: an isomorphic response of guttate psoriasis in a tattoo.
Since Koebner's first observance of the isomorphic response of psoriasis in a tattoo in 1872, a paucity of written reports has appeared in the literature describing this phenomenon, although it appears to be common knowledge among dermatologists. We report a strep-induced guttate psoriasis isomorphic response in a Winnie the Pooh tattoo in a 19-year-old.


The TNF Receptors p55 and p75 Mediate Chemotaxis of PMN Induced by TNFalpha and a TNFalpha 36-62 Peptide.
The present study was performed to examine whether residues 36-62 of TNFalpha contain the chemotactic domain of TNFalpha, and whether the p55 and p75 TNF receptors are involved in TNFalpha induced chemotaxis. The chemotactic effect of TNFalpha on PMN was inhibited by the mAbs Hrt-7b and Utr-1, against the p55 and p75 TNF receptors, respectively. Both receptors may therefore be required for mediating the chemotactic effect of TNFcz. The synthetic TNFalpha 36-62, similar to TNFalpha, had chemotactic effects on both PMN and monocytes. The chemotactic activity of the TNFalpha 36-62 peptide on PMN, was inhibited by Htr-7b, Utr-1 and soluble p55 receptor, which shows that the peptide possessed the ability to induce chemotaxis through the TNF receptors. In contrast to TNFalpha, the peptide did not show a cytotoxic activity against WEHI 164 flbrosarcoma cells. It is suggested that different domains of the TNFalpha molecule induce distinct biological effects.


Endocrine gene neurotensin: molecular mechanisms and a model of intestinal differentiation.
The peptide neurotensin (NT) is an important intestinal hormone that affects gastrointestinal (GI) secretion, motility, and growth. This review describes the effects of NT in the GI tract and the distribution and localization of the gene encoding NT (i.e., NT/N). In addition, the molecular regulation of NT/N gene expression is discussed with special emphasis on the transcription factors, signal transduction pathways, and the role of gene methylation in modulating NT/N gene expression. Our studies have identified the NT/N gene as an excellent molecular model by which to define factors regulating intestinal cell differentiation. The in-depth analysis of the cellular processes leading to the expression of "model" intestinal genes, such as NT/N, can provide better understanding of normal gut development and function as well as defining more clearly the cellular events leading to gut neoplasia.


Identification of novel nonmethylene-interrupted fatty acids, 7E,13E-20:2, 7E,13E,17Z-20:3, 9E,15E,19Z-22:3, and 4Z,9E,15E,19Z-22:4, in Ophiuroidea (brittle star) lipids.
Fatty acids of Ophiuroidea (brittle star) Ophiura sarsi have been investigated by gas-liquid chromatography (GLC). About 2-13% of four unidentified fatty acids were found in total fatty acids from a sample caught at a depth of 1,100 m. Structural analyses were undertaken after partial hydrogenation of their concentrates with hydrazine hydrate and subsequent isolation of the monoenoate products by argentation thin-layer chromatography. The structures of the unidentified fatty acids were determined as 7E,13E-eicosadienoic (20:2), 7E,13E,17Z-eicosatrienoic (20:3), 9E,15E,19Z-docosatrienoic (22:3), and 4Z,9E,15E,19Z-docosatetraenoic (22:4) acids by gas chromatography-mass spectrometry of dimethyl disulfide adducts and GLC of the monoenoates on a polar column. These fatty acids belong to a family of nonmethylene-interrupted (NMI) polyunsaturated fatty acids frequently observed in marine invertebrates and conifer seeds. As far as the authors know, however, these NMI fatty acid types with mixed geometry of ethylenic bonds have not been reported previously.


Lack of reliability in the disruption of cognitive performance following exposure to protons.
A series of three replications were run to determine the reliability with which exposure to protons produces a disruption of cognitive performance, using a novel object recognition task and operant responding on an ascending fixed-ratio task. For the first two replications, rats were exposed to head-only exposures to 1000 MeV/n protons at the NASA Space Radiation Laboratory. For the third replication, subjects were given head-only or whole-body exposures to both 1000 and 150 MeV/n protons. The results were characterized by a lack of consistency in the effects of exposure to protons on the performance of these cognitive tasks, both within and between replications. The factors that might influence the lack of consistency and the implications for exploratory class missions are discussed.


Critical concepts in abdominal injury.
Missed intra-abdominal injuries are among the most frequent causes of potentially preventable trauma deaths. The evaluation and management of abdominal trauma is dependant on multiple factors, including mechanism of injury, location of injury, hemodynamic status of the patient, neurologic status of the patient, associated injuries, and institutional resources.


Peptide-cleaving catalyst selective for melanin-concentrating hormone: Oxidative decarboxylation of N-terminal aspartate catalyzed by Co(III)cyclen.
To provide a firm basis for the new paradigm of drug discovery based on catalysts for oxidative cleavage of N-terminal aspartate (Asp) residues of oligopeptides, oligopeptide-cleaving catalysts were searched by using melanin-concentrating hormone (MCH) as the substrate. MCH is a target for designing drugs to reduce obesity. Catalyst candidates containing the Co(III) complex of cyclen as the catalytic center were prepared by multicomponent condensation reactions. From three kinds of chemical libraries containing about 19,000 catalyst candidates, one compound was identified as the MCH-cleaving catalyst. On incubation with the catalyst, the N-terminal Asp residue of MCH was converted to the pyruvate residue by oxidative decarboxylation. Detailed kinetics analysis revealed the catalytic nature of the action of the catalyst. In addition, the kinetics data indicated that MCH can be cleaved with half-lives of 3 h or less with submicromolar catalyst concentrations if the structure of the catalyst is further improved.


The induction and reversal of topoisomerase II cleavable complexes formed by nuclear extract from the CHO DNA repair mutant, xrs1.
The gamma-ray sensitive CHO cell mutant xrs1 is hypersensitive to antitumour drugs that stabilise DNA topoisomerase II (topoII) cleavable complexes. Sensitivity appears to result from DNA double-strand breaks (DSBs) that persist in xrs1 cells, but not wild-type CHO-K1 cells, following drug removal. One possible explanation for the persistence of DSBs in xrs1 cells is a defect in topoII which reduces its ability to reseal the DSBs associated with cleavable complexes following drug removal. To address this possibility, cleavable complexes formed in vitro by incubating VP16, plasmid DNA and nuclear extract from either CHO-K1 or xrs1 cells were induced to reverse by adding EDTA or salt to the reaction, or by raising the temperature to 65 degrees C, or by dilution of the drug. The fraction of drug-induced cleavable complexes that reversed in these experiments was dependent on how reversal was induced, and ranged from 55 to 95%. However, the extent of reversal was independent of the source of nuclear extract in all of the experiments, indicating that CHO-KI and xrs1 topoII is equally able to reseal complex-associated DSBs during cleavable complex reversal in vitro.


Can Virtual Simulation Replace Clinical Practical Training for Psychiatric Nursing?
In Korea, virtual simulation is currently replacing clinical practical training for psychiatric nursing due to the COVID-19 pandemic. This study aimed to explore the virtual simulation experiences of nursing students through focus-group interviews. Three categories were derived: 'glad that the patients were not real people', 'bridge between the text world and the real world', and 'supplementations needed for virtual simulation to replace clinical practice'. Although virtual simulation helped acquire knowledge and skills related to psychiatric nursing practice in a safe environment, there were limitations to replacing clinical practice. To overcome this, a diverse approach between students and professors is required.


The role and mechanism of myeloperoxidase in dermatomyositis.
Dermatomyositis (DM) is the best known subtype of idiopathic inflammatory myopathies. The hallmarks of DM muscle pathology including microangiopathy, inflammatory infiltration, and perifascicular atrophy. Recent findings have revealed pathogenetic effects of myeloperoxidase (MPO) by causing oxidative damage and regulating abnormal immunity in multiple disease conditions. In this study, we aimed to explore the role of MPO in the pathogenesis of DM. The peripheral blood mononuclear cell (PBMC) mRNA expression and DNA methylation of MPO were verified using real-time qPCR and bisulfite pyrosequencing, respectively. Plasma MPO levels were measured with enzyme-linked immunosorbent assay, and their relationships with clinical characteristics were analyzed. The expression and distribution of MPO in muscle were tested by immunofluorescence. Purified human native MPO protein was used to stimulate human dermal microvascular endothelial cells (HDMECs) and skeletal muscle myotubes. The cell viability, tube forming capacity, permeability, adhesion molecule expressions in HDMECs, and atrophy and programmed cell death pathways in myotubes were then observed. MPO gene methylation was decreased, while mRNA expression and plasma levels were increased in DM. Plasma MPO of DM patients was positively correlated with serum creatine kinase (CK). MPO mainly distributed around endomysia capillaries and perifascicular atrophy in DM muscle biopsies, and was co-localized with CD4+, CD8+ T cells and CD19+ B cells. MPO not only could influence the cell viability, tube forming capacity, permeability and expression of adhesion molecules (including ICAM 1, VCAM 1 and E-selectin) of HDMECs, but also could cause atrophy of myotubes. Our study disclosed, for the first time, that MPO plays an important role in promoting inflammatory infiltration and inducing muscle damage in DM patients. MPO may be a potential biomarker for DM muscle involvement and MPO targeted drugs may be promising in DM treatment.


Potassium channels related to primary aldosteronism: Expression similarities and differences between human and rat adrenals.
Three potassium channels have been associated with primary aldosteronism (PA) in rodents and humans: KCNK3 (TASK-1), KCNK9 (TASK-3), and KCNJ5 (Kir3.4). Mice with deficiency in Kcnk3 and Kcnk9 have elevated aldosterone production and blood pressure. In humans, adrenal tumors with somatic mutations in KCNJ5 cause PA. However, there are very few reports on the expression patterns of these genes in humans versus rodents. Herein, we compared human and rat mRNA expression (by quantitative real-time polymerase chain reaction (qPCR) and protein levels (by immunohistochemistry) across three tissues (adrenal, brain, heart) and two laser-captured adrenal zones (zona glomerulosa, zona fasciculata). Our findings show that expression patterns of KCNK3, KCNK9, and KCNJ5 are inconsistent between rats and humans across both tissues and adrenal zones. Thus, species variation in the expression of PA-related potassium channels indicates an evolutionary divergence in their role in regulating adrenal aldosterone production.


Modulations of collicular visual responses by acoustic stimuli in rabbits.
This study analyzes the influences of an acoustic stimulus upon neuronal light responses of superficial layers of the superior colliculus in anesthetized and paralyzed rabbits. The results have revealed that even if visually-responsive cells fail to be excited by sound, the latter is still capable of modifying light-evoked discharge. The influence may be "short-term" (the discharge rate recovers within 500 ms) or it may be "long-term" (the firing rate remains modified for several seconds). This audio-visual interaction depends upon several factors: the time of occurrence of both stimuli, the physical aspects of the visual target, the relative positions of the speaker and the visual receptive field, and finally, the sensitivity of the unit to movement direction. Data indicates that cells of the most dorsal (hence visual) layers of the superior colliculus are influenced by sound. It is concluded that the colliculus may use the sound as an additional cue to orientate the animal. Also, collicular cells could "memorize" for several seconds various features present in the environment.


Associations between patient and disease characteristics and severe adverse events during immune checkpoint inhibitor treatment: An observational study.
With increasing use of immune checkpoint inhibitors (ICIs) more patients will develop severe and potentially life-threatening immune-related adverse events (irAEs). So far, predictive models for the occurrence of grade ≥3 irAEs are lacking. Therefore, we analysed associations between patient and disease characteristics, and the occurrence of grade ≥3 irAEs. Patients with cancer who were treated with anti-PD-1 (+/-anti-CTLA-4) between July 2015 and February 2020, and who were prospectively included in the MULTOMAB-trial, were eligible for this cohort study. Time to and occurrence of grade ≥3 irAEs according to CTCAE v5.0 were retrospectively registered. The associations between patient and disease characteristics and irAE occurrence were analysed using the competing risk cox-regression model of Fine and Gray. Analyses were performed separately in patients treated with monotherapy (anti-PD-1) and combination therapy (anti-PD-1 + anti-CTLA-4). Subgroup analyses were performed in tumour types with the highest number of patients; melanoma and NSCLC. Out of 641 patients, 106 patients (17%) experienced grade ≥3 irAEs. None of the analysed factors were associated with grade ≥3 irAE occurrence in the monotherapy (n = 550) or the combination therapy (n = 91) groups, nor in the subgroup analyses. Of interest, none of the patients with NSCLC with a WHO performance status of 0 (n = 34) experienced grade ≥3 irAEs. Most common NSCLC histology types were adenocarcinoma (n = 99/55%) and squamous cell carcinoma (n = 39/22%). This study shows that patient and disease characteristics are not able to predict the occurrence of serious AEs in patients treated with ICIs.


Acarbose Mitigates Age-Dependent Alterations in Erythrocyte Membrane Transporters During Aging in Rats.
Acarbose (ACA), a well-studied and effective inhibitor of α-amylase and α-glucosidase, is a postprandial-acting antidiabetic medicine. The membrane of the erythrocyte is an excellent tool for analyzing different physiological and biochemical activities since it experiences a range of metabolic alterations throughout aging. It is uncertain if ACA modulates erythrocyte membrane activities in an age-dependent manner. As a result, the current study was conducted to explore the influence of ACA on age-dependent deteriorated functions of transporters/exchangers, disrupted levels of various biomarkers such as lipid hydroperoxides (LHs), protein carbonyl (PCO), sialic acid (SA), total thiol (-SH), and erythrocyte membrane osmotic fragility. In addition to a concurrent increase in Na+/H+ exchanger activity and concentration of LH, PCO, and osmotic fragility, we also detected a considerable decrease in membrane-linked activities of Ca2+-ATPase (PMCA) and Na+/K+-ATPase (NKA), as well as concentrations of SA and -SH in old-aged rats. The aging-induced impairment of the activities of membrane-bound ATPases and the changed levels of redox biomarkers were shown to be effectively restored by ACA treatment.


Prevention of dentine caries using silver diamine fluoride application followed by Er:YAG laser irradiation: an in vitro study.
To evaluate the preventive effect of Er:YAG laser (EYL) irradiation followed by silver diamine fluoride (SDF) application on dentine with cariogenic biofilm challenge. Twenty-four dentine slices were prepared from extracted sound human third molars. Each slice was cut into four parts for SDF application, followed by EYL irradiation (group SL), SDF application (group S), EYL irradiation (group L) and water (group W). The specimens were subjected to cariogenic biofilm challenge for 12 h, followed by immersion in a buffered remineralising solution containing calcium chloride and sodium hypophosphate for 12 h at 37 °C. Surface morphological changes in the specimens were examined using scanning electronic microscopy. Elemental analysis was performed using energy-dispersive X-ray spectrometry. Micro-mechanical properties were investigated by nano-indentation. The specimen surfaces of groups SL and L showed laser melting contours with narrowed dentinal orifices. Group S showed a partial tubular occlusion. A porous surface was observed in group W, indicating demineralisation. The mean (SD) fluoride weight percentages were 3.93 (0.91), 3.10 (0.61), 0.17 (0.09) and 0.32 (0.07) in groups SL, S, L and W, respectively, (p < 0.001; SL, S > L, W). The mean (SD) micro-hardness values in GPa were 1.84 (0.22), 0.49 (0.13), 0.41 (0.11) and 0.30 (0.06) in groups SL, S, L and W, respectively, (p < 0.001; SL > S > L, W). The mean (SD) elastic moduli in GPa were 75.1 (7.2), 20.0 (1.3), 24.3 (5.2) and 20.2 (2.8) in groups SL, S, L and W, respectively, (p < 0.001; SL > S, L, W). SDF application followed by EYL irradiation on a dentine surface increased its resistance to cariogenic biofilm challenge.


Modelling of the thyroid hormone synthesis as a part of nonlinear reaction mechanism with feedback.
The synthesis of thyroid hormones in the hypothalamic-pituitary-thyroid (HPT) axis was studied. For this purpose, a reaction model for HPT axis with stoichiometric relations between the main reaction species was postulated. Using the law of mass action, this model has been transformed into a set of nonlinear ordinary differential equations. This new model has been examined by stoichiometric network analysis (SNA) with the aim to see if it possesses the ability to reproduce oscillatory ultradian dynamics founded on the internal feedback mechanism. In particular, a feedback regulation of TSH production based on the interplay between TRH, TSH, somatostatin and thyroid hormones was proposed. Besides, the ten times larger amount of produced T4 with respect to T3 in the thyroid gland was successfully simulated. The properties of SNA in combination with experimental results, were used to determine the unknown parameters (19 rate constants of particular reaction steps) necessary for numerical investigations. The steady-state concentrations of 15 reactive species were tuned to be consistent with the experimental data. The predictive potential of the proposed model was illustrated on numerical simulations of somatostatin influence on TSH dynamics investigated experimentally by Weeke et al. in 1975. In addition, all programs for SNA analysis were adapted for this kind of a large model. The procedure of calculating rate constants from steady-state reaction rates and very limited available experimental data was developed. For this purpose, a unique numerical method was developed to fine-tune model parameters while preserving the fixed rate ratios and using the magnitude of the experimentally known oscillation period as the only target value. The postulated model was numerically validated by perturbation simulations with somatostatin infusion and the results were compared with experiments available in literature. Finally, as far as we know, this reaction model with 15 variables is the most dimensional one that have been analysed mathematically to obtain instability region and oscillatory dynamic states. Among the existing models of thyroid homeostasis this theory represents a new class that may improve our understanding of basic physiological processes and helps to develop new therapeutic approaches. Additionally, it may pave the way to improved diagnostic methods for pituitary and thyroid disorders.


Transgenic pigs expressing human decay-accelerating factor regulated by porcine MCP gene promoter.
Porcine membrane cofactor protein (pMCP) is abundantly expressed throughout the body with particularly strong expression on the vascular endothelia. Previous studies demonstrated that the promoter of the pMCP gene induced efficient expression of a human complement regulatory protein, decay-accelerating factor (DAF; CD55), in transgenic mice. In the present study, we tried to produce transgenic pigs with two hybrid genes, 0.9/hDAF and 5.4/hDAF, which were composed of human DAF (hDAF) gene regulated under pMCP promoters of different lengths (0.9 and 5.4 kb). Five live founder transgenic pigs were obtained only with the 0.9/hDAF construct. Although, four founder pigs transmitted the transgene to the second generation, the transmission rates varied among founders. We examined the expression of hDAF in tissues of descendants of two lines (Dm1 and Dm4). Human DAF specific RNAs were confirmed by an RT-PCR analysis in all organs examined. Levels of hDAF protein in the organs from the descendants of Dm1 line were higher than those in the corresponding human organs as determined by enzyme-linked immunosorbent assay. Immunohistochemical studies showed that the tissue distribution of hDAF in the descendants of both lines was similar to that of endogenous pMCP. The expression level of hDAF on the vascular endothelial cells in Dm1 line was twice that on the corresponding human cells. We tested whether proinflammatory cytokines upregulate an efficiency of pMCP promoter on hDAF expression in transgenic pigs. Although the expression of hDAF on the human endothelial cells increased with a combination of cytokines, tumor necrosis factor alpha and interferon-gamma, no cytokine-induced upregulation was seen in the cells of transgenic pigs. The endothelial cells from transgenic pigs exhibited high resistance to the human serum-mediated cytolysis.


Investigation on the adsorption of phosphorus by Fe-loaded ceramic adsorbent.
This aim of this study was to remove phosphorus from aqueous solution using a Fe-loaded ceramic (Fe-LC) adsorbent prepared by mixing dolomite, montmorillonite, FeSO4·7H2O and starch. Simplex-centroid mixture design method was used to determine the optimum mixture proportions by evaluating both phosphorus adsorption efficiency and adsorbent hardness. The study found that the optimum adsorption capacity and the strength can be achieved with the composition of 3.87g dolomite, 3.00g starch, 2.13g montmorillonite and 1.00g FeSO4·7H2O (10g in total). The optimized Fe-LC was evaluated in the batch and the fixed bed experiments. The point of zero charge, pHpzc was found to be 6.0. The adsorption kinetic and isotherm data well agreed with the pseudo-second-order kinetic and the Langmuir isotherm models, respectively. The breakthrough time increased with increasing in the bed depth, whereas inverse relationship was observed with the initial phosphorus concentration in the fixed bed studies. The co-existing anions (SO4(2-), NO3(-) and Cl(-)) had negligible influence on phosphorus removal. The BDST and Thomas model explained the breakthrough behavior for phosphorus removal with a high degree of correlation.


Vertical stratification of °13C values in closed natural and plantation forests in the Luquillo mountains, Puerto Rico.
The variability of δ13C values was measured in leaf, stem and root tissues of several tree species growing in closed natural and plantation forests in the Luquillo mountains of Puerto Rico. Results confirm a significant decrease of δ13C values from the tree canopy to the forest floor. The values measured in understory plants growing in gaps were not significantly different from the average for plants growing under the forest shade. Seedling leaf values tended to be more positive than those of saplings, probably reflecting the contribution of organic matter from the mother tree. Photosynthetic independence on the forest floor results in a reduction in °13C value. Stem and root tissue values of seedlings and saplings were less negative than those of the leaves of the same plants. It is suggested that this difference results from the slower change in isotopic composition experienced by the woody tissue, as the seedlings become photosynthetically independent in the forest floor.


Can transsynaptic viral strategies be used to reveal functional aspects of neural circuitry?
Viruses have proved instrumental to elucidating neuronal connectivity relationships in a variety of organisms. Recent advances in genetic technologies have facilitated analysis of neurons directly connected to a defined starter population. These advances have also made viral transneuronal mapping available to the broader neuroscience community, where one-step rabies virus mapping has become routine. This method is commonly used to identify inputs onto defined cell populations, to demonstrate the quantitative proportion of inputs coming from specific brain regions, or to compare input patterns between two or more cell populations. Furthermore, the number of inputs labeled is often assumed to reflect the number of synaptic connections, and these viruses are commonly believed to label strong synapses more efficiently than weak synapses. While these maps are often interpreted to provide a quantitative estimate of the synaptic landscape onto starter cell populations, in fact very little is known about how transneuronal transmission takes place. We do not know how these viruses transmit between neurons, if they display biases in the cell types labeled, or even if transmission is synapse-specific. In this review, we discuss the experimental evidence against or in support of key concepts in viral tracing, focusing mostly on the use of one-step rabies input mapping and related methods. Does spread of these viruses occur specifically through synaptic connections, preferentially through synapses, or non-specifically? How efficient is viral transneuronal transmission, and is this efficiency equal in all cell types? And lastly, to what extent does viral labeling reflect functional connectivity?


Validation of the care notebook for measuring physical, mental and life well-being of patients with cancer.
To measure patients' QOL in the daily practice of clinical oncology, we developed and tested the Care Notebook. This instrument has 24 questions expressed in single words or short phrases to make it more acceptable to patients. The Care Notebook, EORTC QLQ-C30 and FACIT-Sp-12 were administered to 249 outpatients with cancer. Construct validity was investigated by cluster analysis and multitrait scaling analysis. The results showed that three scales (physical well-being, mental well-being, and life well-being) could explain 55% of the variance in scores. The life well-being scale could be divided into subscales of Daily Functioning, Social Functioning, and Subjective QOL. Multitrait scaling analysis confirmed convergent and discriminant validity of these scales and subscales. Internal consistency and test-retest reliability were favorable. Differences in Care Notebook scores were also consistent with differences in performance status rating (known-groups validity), and Care Notebook scores correlated with EORTC QLQ-C30 and FACT-Sp-12 scores (concurrent validity). The Care Notebook allows clinical oncologists to easily collect valid and reliable QOL information of physical, mental, and life well-being repeatedly and with minimal burden on patients.