The experimental receptivity of Helicella (Helicella) itala and Cepaea nemoralis (Mollusca, Helicidae) to larvae of Muellerius sp. and Neostrongylus linearis (Nematoda, Protostrongylidae) from chamois (Rupicapra rupicapra).
Two batches of Helicella (H.) itala (adult specimen) and two of Cepaea nemoralis (adult and young specimens) were experimentally infected with larvae I (L-I) of Muellerius sp. and Neostrongylus linearis obtained from the lungs and faeces of Rupicapra rupicapra. In assess larval development, the number and percentage of the total number of larvae (L-I + L-II - L-III) per mollusc were studied, together with the number and percentage of L-III per snail and the days on which the different larval stages were reached. The development of Muellerius sp. and N. linearis was greater in larvae from faeces. For both species of molluscs, the values for the percentages of the total number of larvae and L-III were higher in N. linearis than in Muellerius sp., but there were no notable differences in the days on which the various larval stages were reached. Both nematodes achieved a greater degree of development in young specimens of C. nemoralis than in adults. Whether the larvae came from faeces or the lungs, H. (H.) itala was a better intermediate host than C. nemoralis for Muellerius sp. and N. linearis.


Effect of untreated and treated temporomandibular joint arthritis on mandibular volume development in growing rabbits.
The goal of this work was to investigate the volume development of the mandible in growing rabbits with bilaterally induced temporomandibular joint (TMJ) arthritis that was either left untreated or treated with the tumor necrosis factor-alpha (TNF-α) antagonist etanercept. A total of 18 New Zealand White rabbits aged 8 weeks were randomized to three groups of 6 animals each. Two of these groups were used as arthritis groups by sensitizing the 12 animals to ovalbumin (OA) at 10 weeks, followed by intraarticular OA injections to induce bilateral TMJ arthritis and repeating these injections every 3 weeks to maintain the inflammation. One of the two arthritis groups was treated by weekly subcutaneous etanercept injections, whereas the other group was left untreated. The remaining 6 animals served as controls. Maxillofacial CT scans were obtained at 3-week intervals (from week 10 of the rabbits' lives to the end of the experiment at 22 weeks) to volumetrically track the development of the mandibles after segmentation. The mandibles did not grow at a continuous rate, but the rate of development was found to decrease in all groups over the course of the study (weeks 10-22). The most extensive volume increases were noted during weeks 10-13. Severe growth deficiencies, especially of the condylar processes, were observed in the arthritis group not receiving treatment. The arthritis group treated with etanercept showed better rates of growth without, however, reaching the normal range of the control group. Antigen-induced TMJ arthritis was found to involve severe problems of growth similar to those in juvenile idiopathic arthritis. Etanercept can improve the volume development but does not reestablish an entirely normal rate of growth.


Measuring Transcription Rate Changes via Time-Course 4-Thiouridine Pulse-Labelling Improves Transcriptional Target Identification.
Identifying changes in the transcriptional regulation of target genes from high-throughput studies is important for unravelling molecular mechanisms controlled by a given perturbation. When measuring global transcript levels only, the effect of the perturbation [e.g., transcription factor (TF) overexpression or drug treatment] on its target genes is often obscured by delayed feedback and secondary effects until the changes are fully propagated. As a proof of principle, we show that selective measuring of transcripts that are only synthesised after a perturbation [4-thiouridine (4sU) sequencing (4sU-seq)] is a more sensitive method to identify targets and time-dependent transcriptional responses than global transcript profiling. By metabolically labelling RNA in a time-course setup, we could vastly increase the sensitivity of MYCN target gene detection compared to traditional RNA sequencing. The validity of targets identified by 4sU-seq was demonstrated using chromatin immunoprecipitation sequencing and neuroblastoma microarray tumour data. Here, we describe the methodology, both molecular biology and computational aspects, required to successfully apply this 4sU-seq approach.


Alzheimer's Dementia After Exposure to Anesthesia and Surgery in the Elderly: A Matched Natural Experiment Using Appendicitis.
The aim of this study was to determine whether surgery and anesthesia in the elderly may promote Alzheimer disease and related dementias (ADRD). There is a substantial conflicting literature concerning the hypothesis that surgery and anesthesia promotes ADRD. Much of the literature is confounded by indications for surgery or has small sample size. This study examines elderly patients with appendicitis, a common condition that strikes mostly at random after controlling for some known associations. A matched natural experiment of patients undergoing appendectomy for appendicitis versus control patients without appendicitis using Medicare data from 2002 to 2017, examining 54,996 patients without previous diagnoses of ADRD, cognitive impairment, or neurological degeneration, who developed appendicitis between ages 68 through 77 years and underwent an appendectomy (the ''Appendectomy'' treated group), matching them 5:1 to 274,980 controls, examining the subsequent hazard for developing ADRD. The hazard ratio (HR) for developing ADRD or death was lower in the Appendectomy group than controls: HR = 0.96 [95% confidence interval (CI) 0.94-0.98], P < 0.0001, (28.2% in Appendectomy vs 29.1% in controls, at 7.5 years). The HR for death was 0.97 (95% CI 0.95-0.99), P = 0.002, (22.7% vs 23.1% at 7.5 years). The HR for developing ADRD alone was 0.89 (95% CI 0.86-0.92), P < 0.0001, (7.6% in Appendectomy vs 8.6% in controls, at 7.5 years). No subgroup analyses found significantly elevated rates of ADRD in the Appendectomy group. In this natural experiment involving 329,976 elderly patients, exposure to appendectomy surgery and anesthesia did not increase the subsequent rate of ADRD.


Effects of chronic low-dose acrylamide exposure on progressive ratio performance in adolescent rats.
Acrylamide (ACR) is a neurotoxicant known to produce peripheral neuropathy in rats and humans, but little is known of its potential for producing cognitive or motivational alterations. Chronic exposure to low doses of ACR as a food contaminant is known to occur widely in humans. This research evaluated the effects of daily ACR exposure on food-motivated behavior, with exposures beginning prenatally on gestation day 6 and continuing through approximately postnatal day (PND) 85. Plug-positive Fischer 344 dams (9-10 per dose) were gavaged daily with 0, 0.1, 0.3, 1.0 or 5.0mg/kg/day ACR. On PNDs 1-22, pups were gavaged with the same dose their dam had received. On PND 22, pups were weaned and pair-housed with a same-sex littermate and ACR exposure continued at 0, 1, 3, 10 and 50ppm via drinking water. One male and one female pup per litter were tested in an operant chamber under a progressive ratio (PR) schedule of food reinforcement from approximately 6 to 12 weeks of age. Results over 6 weeks of testing indicated a significant treatment effect of ACR on number of reinforcers earned, with Tukey HSD post hoc tests revealing significantly fewer reinforcers earned in the 5.0mg/kg/day dose group than in controls. A significant effect of ACR on response rate was also observed, with the Tukey HSD post hoc tests revealing a significantly lower response rate in the 5.0mg/kg/day group than in controls. No effects of ACR were observed on post-reinforcement pause. These data suggest that daily ACR exposure at 5.0mg/kg/day can produce measurable decrements on aspects of food-motivated behavior.


[Informed trust in science: lessons from the COVID 19 pandemic for the conceptualization of science literacy].
Informed trust in science is necessary for the 'interfaces' within the flow of knowledge between citizens' everyday understanding of the pandemic and the dynamically evolving state of knowledge in the sciences. This is the core thesis of this paper. Without science, the COVID-19 pandemic can neither be understood nor controlled, and for this to happen, citizens must engage with science based knowledge. However, such knowledge is dynamic (evolving and intertwined with normative issues). Furthermore, science based knowledge competes with pseudoscientific contributions. As non-experts, laypersons must therefore decide whom to trust. The paper describes the concept of functional scientific literacy as a prerequisite of informed trust. The knowledge bases for judgments of informed trust should be taught in school and judging rationally about the trustworthiness of science-related knowledge claims should be practiced.


Retraction Note: Endoplasmic reticulum stress triggers Xanthoangelol-induced protective autophagy via activation of JNK/c-Jun Axis in hepatocellular carcinoma.
An amendment to this paper has been published and can be accessed via the original article.


Release of dialkylglycerol from purple membrane phospholipids by phospholipase D.
When the major polar lipid of purple membrane, a dialkyl analogue of phosphatidyl glycerophosphate, is treated with phospholipase D under the usual assay conditions for this enzyme, the reaction yields dialkylglycerol and glycerol bisphosphate, i.e. the kind of products that would be expected from a phospholipase C reaction. The effect is seen both in native purple membranes and with the pure phospholipid in the form of liposomes. The specific activity and kinetic parameters Km and Vmax of phospholipase D for the purple membrane phospholipid are similar to those for egg phosphatidylcholine. The presence of phospholipase C impurities in the phospholipase D preparations has been ruled out as an explanation for the above observations. A hypothesis is suggested, taking into account the peculiar headgroup structure of the bacterial lipid, to explain the seemingly anomalous enzyme behavior.


Energy transfer from photosystem II to photosystem I in Porphyridium cruentum.
Rates of photooxidation of P-700 by green (560 nm) or blue (438 nm) light were measured in whole cells of porphyridium cruentum which had been frozen to -196 degrees C under conditions in which the Photosystem II reaction centers were either all open (dark adapted cells) or all closed (preilluminated cells). The rate of photooxidation of P-700 at -196 degrees C by green actinic light was approx. 80% faster in the preilluminated cells than in the dark-adapted cells. With blue actinic light, the rates of P-700 photooxidation in the dark-adapted and preilluminated cells were not significantly different. These results are in excellent agreement with predictions based on our previous estimates of energy distribution in the photosynthetic apparatus of Porphyridium cruentum including the yield of energy transfer from Photosystem II to Photosystem I determined from low temperature fluorescence measurements.


Current status of education and treatment resources for lymphedema.
Secondary lymphedema (LE) resulting from breast cancer surgery has continued to be an ignored medical diagnosis in U.S. medicine. Subsequently, the majority of women/men undergoing axillary lymph node dissection have not received education in pre- or postoperative LE prevention, causing hundreds of thousands of patients to develop LE. As a result of this ignorance, once the LE develops, these patients receive no or harmful treatment. Over the last decade, the National Lymphedema Network (NLN) has created awareness in the medical community, among patients, and in the general public through information dissemination, educational materials, a national conference, and the activism of patients and a small group of concerned professionals. Complete decongestive physiotherapy (CDP), which includes manual lymph drainage, is a successful treatment for LE that has now been introduced. Slowly, the U.S. medical community is beginning to recognize and support this condition and its treatment, and secondary LE has now become an acceptable diagnosis (ICD9-457.0). Subsequently, due to patient and medical activism and the work of the NLN, Medicare now covers 2 weeks of CDP treatment (in Florida), and two landmark bills (AB-12 and S-609) that will cover LE treatment for postbreast carcinoma patients are currently pending (as of February 22, 1998). In the last decade, the NLN has developed a strong foundation of knowledge and has planted LE on the medical map in the United States. With collaboration between the NLN and all breast carcinoma groups and LE specialists, it is hoped that lymphology will become a standard course of study in medical schools across the country and that LE will become a household word, with proper treatment available for postbreast carcinoma LE patients in the United States.


Prognostic factors for gastric cancer influencing clinical practice.
Despite a slow decline in the incidence of gastric cancer over the last 90 years, we can still expect to see over 100,000 patients die of this disease each decade in England and Wales. The 5-year survival rate has not improved during this century, which is largely due to the stage at diagnosis being unchanged. There are a number of prognostic determinants in gastric cancer which have clinical relevance. Age is an important determinant; patients under 40 years commonly have more advanced diffuse lesions than older patients so that a higher index of suspicion needs to be maintained in younger patients with persistent symptoms. Conflicting reports make it unclear whether the duration of symptoms bears any relationship to tumor stage, but there is some evidence that actively shortening the symptom duration by early investigation can have a beneficial effect in the proportion of patients diagnosed with early cancers. The site of the tumor is important; unfortunately, the proportion of patients with cardia lesions is increasing and this has had the effect of reducing the overall survival. Tumor size should not play a part in the decision to resect a lesion as most studies show no clear relationship between tumor size and stage. Tumor stage is the most important prognostic determinant and efforts to increase the proportion of stage I cancers presenting for surgery can be shown to alter the natural history of the disease, by diagnosing it when it is still surgically curable.


Identification of New Specificity Determinants in Bacterial Purine Nucleobase Transporters based on an Ancestral Sequence Reconstruction Approach.
The relation of sequence with specificity in membrane transporters is challenging to explore. Most relevant studies until now rely on comparisons of present-day homologs. In this work, we study a set of closely related transporters by employing an evolutionary, ancestral-reconstruction approach and reveal unexpected new specificity determinants. We analyze a monophyletic group represented by the xanthine-specific XanQ of Escherichia coli in the Nucleobase-Ascorbate Transporter/Nucleobase-Cation Symporter-2 (NAT/NCS2) family. We reconstructed AncXanQ, the putative common ancestor of this clade, expressed it in E. coli K-12, and found that, in contrast to XanQ, it encodes a high-affinity permease for both xanthine and guanine, which also recognizes adenine, hypoxanthine, and a range of analogs. AncXanQ conserves all binding-site residues of XanQ and differs substantially in only five intramembrane residues outside the binding site. We subjected both homologs to rationally designed mutagenesis and present evidence that these five residues are linked with the specificity change. In particular, we reveal Ser377 of XanQ (Gly in AncXanQ) as a major determinant. Replacement of this Ser with Gly enlarges the specificity of XanQ towards an AncXanQ-phenotype. The ortholog from Neisseria meningitidis retaining Gly at this position is also a xanthine/guanine transporter with extended substrate profile like AncXanQ. Molecular Dynamics shows that the S377G replacement tilts transmembrane helix 12 resulting in rearrangement of Phe376 relative to Phe94 in the XanQ binding pocket. This effect may rationalize the enlarged specificity. On the other hand, the specificity effect of S377G can be masked by G27S or other mutations through epistatic interactions.


Drug-induced regulatory overcompensation has motivational consequences: Implications for homeostatic and allostatic models of drug addiction.
Initial administration of 60% nitrous oxide (N2O) at 21°C ambient temperature (Ta) reduces core temperature (Tc) in rats, but tolerance develops to this hypothermic effect over several administrations. After additional N2O administrations, a hyperthermic overcompensation (sign-reversal) develops such that Tc exceeds control levels during N2O inhalation. This study investigated whether rats would employ behavioral thermoregulation to facilitate, or oppose, a previously acquired hyperthermic overcompensation during N2O administration. To establish a hyperthermic sign-reversal, male Long-Evans rats (N=12) received ten 3-h administrations of 60% N2O while housed in a gas-tight, live-in, "inactive" thermal gradient (~21°C). Following the tenth N2O exposure, the thermal gradient was activated (range of 10-37°C), and rats received both a control gas session and a 60% N2O test session in counterbalanced order. Mean Tc during N2O inhalation in the inactive gradient was reliably hypothermic during the first exposure but was reliably hyperthermic by the tenth exposure. When subsequently exposed to 60% N2O in the active gradient, rats selected a cooler Ta, which blunted the hyperthermic sign-reversal and lowered Tc throughout the remainder of the N2O exposure. Thus, autonomic heat production effectors mediating the hyperthermia were opposed by a behavioral effector that promoted increased heat loss via selection of a cooler ambient temperature. These data are compatible with an allostatic model of drug addiction that suggests that dysregulatory overcompensation in the drugged-state may motivate behaviors (e.g., drug taking) that oppose the overcompensation, thereby creating a vicious cycle of escalating drug consumption and recurring dysregulation.


Abnormal heart rate characteristics before clinical diagnosis of necrotizing enterocolitis.
Earlier diagnosis and treatment of necrotizing enterocolitis (NEC) in preterm infants, before clinical deterioration, might improve outcomes. A monitor that measures abnormal heart rate characteristics (HRC) of decreased variability and transient decelerations was developed as an early warning system for sepsis. As NEC shares pathophysiologic features with sepsis, we tested the hypothesis that abnormal HRC occur before clinical diagnosis of NEC. Retrospective review of Bells stage II to III NEC cases among infants <34 weeks gestation enrolled in a prospective randomized clinical trial of HRC monitoring at three neonatal intensive care units. Of 97 infants with NEC and HRC data, 33 underwent surgical intervention within 1 week of diagnosis. The baseline HRC index from 1 to 3 days before diagnosis was higher in patients who developed surgical vs medical NEC (2.06±1.98 vs 1.22±1.10, P=0.009). The HRC index increased significantly 16 h before the clinical diagnosis of surgical NEC and 6 h before medical NEC. At the time of clinical diagnosis, the HRC index was higher in patients with surgical vs medical NEC (3.3±2.2 vs 1.9±1.7, P<0.001). Abnormal HRC occur before clinical diagnosis of NEC, suggesting that continuous HRC monitoring may facilitate earlier detection and treatment.


In situ sequestration of endogenous PDGF-BB with an ECM-mimetic sponge for accelerated wound healing.
Recapitulating the typical features of extracellular matrix (ECM) in engineered biomaterials is crucial for preparing a suitable niche to activate endogenous tissue repair. Here, we report the design and evaluation of an ECM-mimetic scaffold that successfully accelerated wound healing through enriching endogenous platelet-derived growth factor-BB (PDGF-BB). Specifically, we prepared a electrospun hydrogel sponge (EGS) comprising a PDGF-BB-binding polysaccharide (EUP3) and gelatin. The two polymers in concert exerted a 'retention-and-release' function: upon the application of EGS in vivo, EUP3 started to bind and sequester endogenous PDGF-BB at the wound site; gradually, gelatin was degraded to free the PDGF-BB/EUP3 complex that acted on the cells in situ. Our serial in vitro and in vivo tests validated the efficacy of EGS in retaining PDGF-BB, releasing PDGF-BB/EUP3 in response to collagenase, and promoting various PDGF-BB-mediated regenerative activities. Particularly, EGS accelerated the repair of a full-thickness skin wound in mice and induced optimal neo-tissue formation, without the addition of any exogenous GFs, cells or genes. Collectively, our results suggest that, by mimicking the distinctive GF-affinitive feature of ECM, EGS as an engineered biomaterial can effectively harness the endogenous regenerative power of the native tissue. Our investigation may inspire the design of new, effective and safer approaches for tissue regeneration.


Advanced Lung Cancer Inflammation Index Predicts Outcomes of Patients With Colorectal Cancer After Surgical Resection.
The advanced lung cancer inflammation index is considered a useful prognostic biomarker of clinical outcomes in patients with malignancies. However, the prognostic value of the advanced lung cancer index in patients with colorectal cancer who underwent surgical resection remains unclear. In this study, we evaluated the prognostic value of the advanced lung cancer index in patients with colorectal cancer. Prospectively obtained data of patients with colorectal cancer were retrospectively evaluated to clarify the clinical relevance of the advanced lung cancer index. We conducted this study at a single expert center. We enrolled 298 patients with colorectal cancer who underwent surgical resection in this retrospective study. The primary outcome was the clinical relevance of the advanced lung cancer index in patients with rectal cancer. Low status of advanced lung cancer index was significantly correlated with undifferentiated histology (p = 0.004), T stage progression (p < 0.001), R1/R2 resection for primary surgery (p = 0.004), and distant metastasis (p < 0.001). Multivariate analysis showed that low advanced lung cancer index status was an independent prognostic factor for both overall survival (HR = 3.21 (95% CI, 1.97-5.19); p < 0.001) and disease-free survival (HR = 2.13 (95% CI, 1.23-3.63); p = 0.008) in patients with colorectal cancer. Furthermore, the clinical burden of the advanced lung cancer index was consistent between sexes, and its prognostic value was verified in patients with clinically relevant stage III colorectal cancer. The present study had several limitations, including retrospective observation and a small sample size of Japanese patients from a single institution. The advanced lung cancer index could be a useful prognostic indicator of clinical outcomes in patients who underwent surgical resection for colorectal cancer. See Video Abstract at http://links.lww.com/DCR/B267. EL ÍNDICE AVANZADO DE INFLAMACIÓN DEL CÁNCER DE PULMÓN, PREDICE LOS RESULTADOS DE LOS PACIENTES CON CÁNCER COLORRECTAL DESPUÉS DE LA RESECCIÓN QUIRÚRGICA: El índice avanzado de inflamación del cáncer de pulmón, es considerado como un útil biomarcador pronóstico, en los resultados clínicos de pacientes con neoplasias malignas. Sin embargo, aún no está claro el valor pronóstico del índice avanzado de cáncer de pulmón, en pacientes con cáncer colorrectal sometidos a resección quirúrgica.Evaluar el valor pronóstico del índice avanzado del cáncer de pulmón, en pacientes con cáncer colorrectal.Los datos obtenidos prospectivamente de pacientes con cáncer colorrectal, fueron evaluados retrospectivamente, para aclarar la relevancia clínica del índice avanzado del cáncer de pulmónEstudio realizado en un solo centro experto.Estudio retrospectivo, incluyendo 298 pacientes con cáncer colorrectal, sometidos a resección quirúrgica.El resultado primario fue la relevancia clínica del índice avanzado de cáncer de pulmón, en pacientes con cáncer rectal.Un índice avanzado de cáncer de pulmón bajo, se correlacionó significativamente con la histología indiferenciada (p = 0.004), la progresión de la etapa T (p <0.001), la resección R1 / R2 para cirugía primaria (p = 0.004) y la metástasis a distancia (p <0.001). El análisis multivariante mostró que el índice avanzado de cáncer de pulmón bajo, era un factor pronóstico independiente, tanto para la supervivencia general (HR = 3.21 IC 95% 1.97-5.19 p <0.001) como para la supervivencia libre de enfermedad (HR = 2.13, IC 95% 1.23-3.63, p = 0,008), en pacientes con cáncer colorrectal. Además, la carga clínica del índice avanzado de cáncer de pulmón, fue consistente entre los sexos y su valor pronóstico se verificó clínicamente relevante, en pacientes con cáncer colorrectal en estadio III.El presente estudio tuvo varias limitaciones, incluyendo la observación retrospectiva y la pequeña muestra de pacientes japoneses, en una sola institución.El índice avanzado de cáncer de pulmón, podría ser un indicador pronóstico útil, en los resultados clínicos de pacientes sometidos a resección quirúrgica por cáncer colorrectal. Consulte Video Resumen http://links.lww.com/DCR/B267.


A 3-week schedule of gemcitabine plus cisplatin as induction chemotherapy for Stage III non-small cell lung cancer.
Our aim was to explore the activity and feasibility of gemcitabine plus cisplatin as induction chemotherapy in patients with Stage IIIA N(2) and selected IIIB non-small cell lung cancer (NSCLC). From September 1997 to July 2000, 70 chemonaive patients with Stage III NSCLC, median age of 64 years, World Health Organization performance status 0, 1, or 2, and the ability to tolerate a pneumonectomy entered the study and received gemcitabine 1250 mg/m(2) on days 1 and 8 and cisplatin 70 mg/m(2) on day 2 every 3 weeks. After three cycles of induction chemotherapy, patients underwent resection or radiotherapy. Responses were seen in 40 of the 69 assessable patients, for an intent-to treat overall response rate of 57.1% (95% confidence interval, 45-62%), with 4.2% complete response. Response rates were 68 and 35% in patients with Stage IIIA and IIIB disease, respectively. The overall pathological CR rate after induction chemotherapy was 3%, with an overall pathological downstaging rate of 20%. Median survival for all patients was 14.5 months, with an estimated 1-year survival rate of 67% (95% CI, 54.3-79.5%). The estimated time to treatment failure was 12.6 months. Grade 3/4 thrombocytopenia was the main hematologic toxicity, occurring in 26% of patients, but was not associated with life-threatening bleeding. Febrile neutropenia was rare and other severe non-hematologic toxicities were uncommon. The 3-week schedule of gemcitabine plus cisplatin is highly active as induction chemotherapy in Stage IIIA N(2) unresectable NSCLC. This suggests a need for a multimodality approach upfront, such as concurrent chemoradiation therapy, particularly in patients with Stage IIIB disease.


Strain specific respiratory air space enlargement in aged rats.
We studied lung structure and function in Fischer-344 and Sprague Dawley rats to compare the pathophysiologic features of the aged lung in animal strains. Both strains were maintained under identical conditions of minimal exposure to injurious environmental agents. We measured the number, size, and surface area of alveoli, pressure-volume characteristics and connective tissue content of lungs at midlife (12 or 14 months of age) and old age (24 months of age). Results showed differences in the older versus younger group of the Sprague Dawley strain as indicated by enlarged air spaces [154 +/- 21 (SEM) versus 118 +/- 13 micromicroliter] (p less than 0.05), increased collagen (hydroxyproline content 4.1 +/- 0.1 versus 3.0 +/- 0.1 mg/lung) (p less than 0.05), and a leftward shifted pressure-volume curve. There was no change in surface area or alveolar number. The structural lesions are consistent with air space enlargement with fibrosis and not emphysema. In contrast, no major changes were found in the lungs with age in Fischer-344 rats. We hypothesize that in the Sprague Dawley strain the aging process impairs the ability of the lung to maintain normal structure and function. Two strains of rats which differ pathologically in old age may be useful in the study of the effects of aging on the lung.


Plasminogen activator inhibitor-1 4G4G genotype is associated with myocardial infarction but not with stable coronary artery disease.
A case control study was conducted to test the hypothesis that plasminogen activator inhibitor type-1 (PAI-1) 4G/5G gene polymorphism confers an increased risk for myocardial infarction (MI) in patients with known coronary atherosclerosis. One hundred fifty-six consecutive patients who presented with acute MI and 111 stable coronary artery disease (SCAD) patients with documented critical coronary artery stenoses were prospectively enrolled. PAI-1 4G/5G gene polymorphism and conventional atherosclerotic risk factors were studied in all patients. PAI-1 4G/5G gene polymorphism was studied in another 281 healthy blood bank donors. The frequency 4G4G genotype was significantly higher in the MI group as compared to SCAD group (32.7% vs. 15.3%, P = 0.001) while it was not statistically significant between MI and healthy control groups (32.7% vs. 26.0%, P = 0.136). Comparing with healthy controls SCAD group had significantly lower frequency of 4G4G genotype (P = 0.024). In comparison with SCAD group PAI-1 4G/4G genotype, male sex and smoking habits favored to MI in univariate analysis with a P value of less than 0.2. These variables were included in multivariate regression model to estimate the associated risk for MI. PAI-1 4G/4G genotype was the only independent variable (OR 2.67, 95%CI 1.43-4.96, P = 0.002) associated with MI in this regression model. Comparing with healthy control group 4G4G genotype was not associated with MI (OR 1.38, 95%CI 0.90-2.12). However, presence of 4G4G genotype had a protective effect against development of SCAD (OR 0.52, 96%CI 0.29-0.92). Compared to patients with critical coronary stenoses, PAI-1 4G/4G genotype was found to be an independent predictor for development of MI in this population. PAI-1 4G4G genotype have a protective effect against development of high grade stable coronary stenoses.


Reduced sCD36 following weight loss corresponds to improved insulin sensitivity, dyslipidemia and liver fat in obese children.
Childhood obesity is a major health problem with serious long-term metabolic consequences. CD36 is important for the development of obesity-related complications among adults. We aimed to investigate circulating sCD36 during weight loss in childhood obesity and its associations with insulin resistance, dyslipidemia, hepatic fat accumulation and low-grade inflammation. The impact of a 10-week weight loss camp for obese children (N=113) on plasma sCD36 and further after a 12-month follow-up (N=68) was investigated. Clinical and biochemical data were collected, and sCD36 was measured by an in-house assay. Liver fat was estimated by ultrasonography and insulin resistance by the homeostasis model assessment (HOMA-IR). Along with marked weight loss, sCD36 was reduced by 21% (P=0.0013) following lifestyle intervention, and individual sCD36 reductions were significantly associated with the corresponding decreases in HOMA-IR, triglycerides and total cholesterol. The largest sCD36 decrease occurred among children who reduced HOMA-IR and liver fat. After 12 months of follow-up, sCD36 was increased (P=0.014) and the metabolic improvements were largely lost. Weight-loss-induced sCD36 reduction, coincident with improved insulin resistance, circulating lipids and hepatic fat accumulation, proposes that sCD36 may be an early marker of long-term health risk associated with obesity-related complications.


Training anesthesiology residents in providing anesthesia for awake craniotomy: learning curves and estimate of needed case load.
To measure the learning curves of residents in anesthesiology in providing anesthesia for awake craniotomy, and to estimate the case load needed to achieve a "good-excellent" level of competence. Prospective study. Operating room of a university hospital. 7 volunteer residents in anesthesiology. Residents underwent a dedicated training program of clinical characteristics of anesthesia for awake craniotomy. The program was divided into three tasks: local anesthesia, sedation-analgesia, and intraoperative hemodynamic management. The learning curve for each resident for each task was recorded over 10 procedures. Quantitative assessment of the individual's ability was based on the resident's self-assessment score and the attending anesthesiologist's judgment, and rated by modified 12 mm Likert scale, reported ability score visual analog scale (VAS). This ability VAS score ranged from 1 to 12 (ie, very poor, mild, moderate, sufficient, good, excellent). The number of requests for advice also was recorded (ie, resident requests for practical help and theoretical notions to accomplish the procedures). Each task had a specific learning rate; the number of procedures necessary to achieve "good-excellent" ability with confidence, as determined by the recorded results, were 10 procedures for local anesthesia, 15 to 25 procedures for sedation-analgesia, and 20 to 30 procedures for intraoperative hemodynamic management. Awake craniotomy is an approach used increasingly in neuroanesthesia. A dedicated training program based on learning specific tasks and building confidence with essential features provides "good-excellent" ability.


Do reactions after whole blood donation predict syncope on return donation?
Adverse reactions after whole blood donation reduce the likelihood of a subsequent donation. Still, many donors return to give blood even after experiencing a reaction. Consequently, we evaluated the risk of recurrent syncope among returning donors. Allogeneic whole blood donors in 2009 who had vasovagal-type reactions including syncope were evaluated for return donation within 12 months and subsequent reactions, based on donation status (novice [first-time] or active [repeat]) or age at index donation. Syncope after a first whole blood donation significantly reduced the frequency of return donation (18%), compared to either presyncopal symptoms (27%; p < 0.0001) or no reaction (35%; p < 0.0001). Among novice donors who returned to donate, syncope was more likely among donors who had any reaction (0.8%) or syncope (3.5%) at their first donation, compared to donors who had no reaction (0.3%; p < 0.0001). Syncope at a first donation identified only 2% (19 of 1062) of syncopal reactions among returning donors. For active, repeat donors who experienced syncope in 2009, a history of prior reactions had no effect on the likelihood of return donation or recurrent syncope. Donation experience strongly influences the likelihood of return donation and the risk of subsequent reactions, but a prior reaction after whole blood donation does not reliably predict recurrent syncope among returning donors.


Corrigendum to "Site directed mutagenesis as a precision tool to enable synthetic biology with engineered modular polyketide synthases" [Synth Syst Biotechnol 5 (2) (2020) 62-80].
[This corrects the article DOI: 10.1016/j.synbio.2020.04.001.].


Ischemic-reperfusion of unilateral external iliac artery in rat: A new model for vasculitic femoral neuropathy.
Clinically, ischemic environment during gynecological surgery at lithotomy position is most common causative factor for the development of vasculitic femoral neuropathy (VFN). The present study was designed to induce the clinically relevant rat model of VFN by ischemic-reperfusion (I/R) injury of unilateral external iliac artery (uEIA). The VFN was induced by 3, 4 and 5h occlusion of uEIA followed by reperfusion. The I/R of uEIA induced VFN was evaluated by (i) behavioral parameters i.e., hind limb temperature; weight bearing capacity; (ii) kinematic analysis i.e., paw posture, splay angle, static sciatic index (SSI), and ankle-angle tests; (iii) evaluation of pain perception i.e., plantar and pin prick; (iv) serum biochemical estimation i.e., nitrate, lipid peroxidation, TNF-α and calcium level; (v) evaluation of motor and sensory nerve conduction velocity; and (vi) measurement of nerve fiber density. The 4 and 5h occlusion of uEIA has produced the potential changes in behavioral, functional, electrophysiological, biochemical and histopathological assessment. The 5h occlusion of uEIA has shown to produce the mortality. Whereas, 3h occlusion does not produce the significant changes in the development of VFN. The 4h ischemic occlusion of uEIA has shown potential rat model of VFN due to its close mimicking capacity of VFN in human. Therefore, it can be useful to explore the newer anti-neuralgic medicine and with their pharmacodynamic action in the field of various neurovascular disorders.


Rapid development of cardiac dysfunction in a canine model of insulin resistance and moderate obesity.
The worldwide incidence of obesity and diabetes continues to rise at an alarming rate. A major cause of the morbidity and mortality associated with obesity and diabetes is heart disease, yet the mechanisms that lead to cardiovascular complications remain unclear. We performed cardiac MRI to assess left ventricular morphology and function during the development of moderate obesity and insulin resistance in a well-established canine model (n = 26). To assess the influence of dietary fat composition, we randomised animals to a traditional lard diet (rich in saturated and monounsaturated fat; n = 12), a salmon oil diet (rich in polyunsaturated fat; n = 8) or a control diet (n = 6). High-fat feeding with lard increased body weight and fasting insulin and markedly reduced insulin sensitivity. Lard feeding also significantly reduced left ventricular function, evidenced by a worsening of circumferential strain and impairment in left ventricular torsion. High-fat feeding with salmon oil increased body weight; however, salmon oil feeding did not impair insulin sensitivity or cardiac function. These data emphasise the importance of dietary fat composition on both metabolic and cardiac function, and have important implications for the relationship between diet and health.


T-DM1-induced thrombocytopenia in breast cancer patients: New perspectives.
Human epidermal growth factor receptor 2 (HER2) is overexpressed in 15-20% of patients with breast cancer. HER2 overexpression is the result of a genetic alteration and this marker is associated with poor clinical outcomes. HER2-targeted therapy can significantly improve the prognosis of patients with either early or advanced HER2-positive breast cancer. One such therapy is the antibody drug conjugate (ADC) trastuzumab emtansine (T-DM1), a combination of trastuzumab and the cytotoxic antimicrotubule agent DM1. After T-DM1 binds HER2, DM1 is subsequently released into the cell. T-DM1 is generally well tolerated and has a relatively low incidence of adverse events. However, there are clinical concerns regarding T-DM1-induced high-grade thrombocytopenia. Here, we summarize the incidence of thrombocytopenia from several clinical trials and review experimental studies to explore the causes for T-DM1-induced thrombocytopenia. Progress in several other ADCs targeting HER2-positive breast cancer was also reviewed. We conclude that T-DM1 uptake by megakaryocytes occurs through either Fcγ receptor binding or through pinocytosis, and we suggest several methods through which these processes could be interrupted to potentially improve the clinical safety of T-DM1. More generally, we recommend that toxicity should be carefully addressed during the development of ADCs.


Inactivation of bladder tumor cells and enzymes by methylene blue plus light.
Using a cystoscopic light source and methylene blue as the sensitizing dye, photoactivation was examined in two types of experiments. In the first, the in vitro study destruction of two enzymes (glucose-6-phosphate dehydrogenase and lactic dehydrogenase) was examined in suspensions of whole and homogenized tumor cells from a transplantable bladder tumor. In the second or in vivo study rats were used to demonstrate that tumor cell suspensions treated with methylene blue plus light, when inoculated into susceptible rats, failed to "take" and produce new tumors. These experiments suggest a possible therapeutic use in treatment of human bladder tumors, though further study would be required.


Neuromodulation for Apathy in Alzheimer's Disease: A Double-Blind, Randomized, Sham-Controlled Pilot Study.
Apathy, a profound loss of motivation, initiation, and goal directed cognition, is a common comorbidity of Alzheimer's disease (AD). The presence of apathy is associated with rapid progression of AD, long-term impairment, disability, and higher mortality. Pharmacological treatments of apathy are limited. The primary objective was to evaluate the efficacy of repetitive transcranial magnetic stimulation (rTMS) for apathy in AD. A randomized, double-blind, parallel-arm, sham-controlled pilot study was conducted in subjects with AD and apathy (N = 20). Subjects were randomized to rTMS or sham treatment (5 days/week) for four weeks. Primary outcome, apathy evaluation scale-clinician version (AES-C), and secondary outcome measures, modified-Mini Mental State Examination (3MS), instrumental activities of daily living (IADL), and clinical global impression (CGI), were assessed at baseline and four weeks. Follow-up visits were conducted at 8 and 12 weeks to test the durability of effects of intervention. Mean age was 77.3 (±7.2) years, 80% were Caucasians and 10% were females. After adjusting for baseline, there was a significantly greater improvement in the AES-C with rTMS compared to sham treatment (-10.1 (-15.9 to -4.3); t (16) = -3.69; p = 0.002) at 4 weeks. There was also significantly greater improvement in 3MS (6.9 (1.7 to 12.0); t (15) = 2.85; p = 0.012), IADL (3.4 (1.0 to 5.9); χ21 = 7.72; p = 0.006), CGI-S (1.4 (0.5 to 2.3), t (16) = 3.29; p = 0.005), and CGI-I (-2.56 (-3.5 to -1.6), t (17) = -5.72; p < 0.001) for rTMS compared to the sham at 4 weeks. The effects of rTMS were durable at 12 weeks. rTMS may be safely used in subjects with AD and may improve apathy, function, and some aspects of cognition.


Fluticasone furoate/Vilanterol 92/22 μg once-a-day vs Beclomethasone dipropionate/ Formoterol 100/6 μg b.i.d.: a 12-week cost analysis in mild-to-moderate asthma.
Asthma is a disease with high cost for the National Health Service. Two of the most recent LABA/ICS combinations for persistent bronchial asthma are Beclomethasone dipropionate/Formoterol (B/F) delivered via the Nexthaler device and Fluticasone furoate/Vilanterol (F/V) delivered via the Ellipta device. No comparison has been carried out yet in terms of cost analysis in asthma, to our knowledge. Aim of the present monocentric, observational, retrospective study was to calculate and compare the costs of mild-to-moderate asthma patients assuming B/F 100/6 μg b.i.d. to those of patients assuming F/V 92/22 μg once-a-day over a 12-week treatment period from the Italian National Health Service perspective. Data were obtained automatically and anonymously from the institutional database of the Lung Unit of the Specialist Medical Centre (CEMS), Verona, Italy, UNI EN ISO 9001-2008 validated. FEV1 values, number of relapses, healthcare resources as hospitalizations due to asthma relapses, days of hospitalization, general practitioner (GP), specialist visits, and days of inactivity, were recorded over the study period together with the use of extra medications (systemic steroids and antibiotics). In order to compare the outcomes achieved in both groups, the propensity score matching method was used in STATA, and statistical significance was accepted for p < 0.05. Clinical data of 77 patients treated with B/F b.i.d (Group A) and of 40 patients treated with F/V 92/22 μg once-a-day (Group B) were selected. The PS-matching process, designed as matching on the baseline covariates, gender, age, FEV1 and comorbidities, returned a cohort of 40 group A patients of the entire cohort matched with 40 patients of group B, fully comparable for demographics and clinical characteristics. In the PS-matched cohort, the mean (±SE) number of relapses per patient during the follow-up was 0.53 (±0.12) in group A and 0.28 (±0.07) in group B. In group A, n = 25 (62.50 %), n = 9 (22.50 %), and n = 6 (15 %) patients had 0, 1, 2 relapses, respectively. In group B, n = 29 (72.50 %), and n = 11 (27.50 %) had 0 and 1 relapse, respectively. Over the study period, the average number of hospitalizations per patient was 0.15 (±0.06), with 0.28 (±0.12) days of hospitalization in group A, and 0.08 (±0.04) with 0.08 (±0.04) days of hospitalization in group B, respectively. The difference between the two groups in terms of FEV1(L) improvement vs baseline was 0.11 in favour of group B (p = 0.007). When results were compared, the improvement in lung function obtained in group B proved significantly higher both in terms of absolute FEV1 and of FEV1 % predicted. The mean (±SE) cost of hospitalizations per patient was € 345.30 (±133.23) in group A and € 172.65 (±98.18) in group B, respectively, with a mean not significant difference of - € 172.65 in favour of group B (p = 0.9). In particular, the mean (±SE) cost for visits per patient was € 26.82 (±3.73) in group A and € 11.36 (±2.30) in group B (p = 0.002), and the mean cost for rescue medications per patient was € 35.24 (±6.93) in group A, and € 18.73 (±3.38) in group B, respectively (p = 0.05). Even if both ICS/LABA combinations were checked over a limited period of time, they seem characterized by a different profile in terms of effect on lung function and economic impact on mild-to-moderate asthma. The once-daily inhalation of combined Fluticasone furoate/Vilanterol 92/22 μg showed the potential for enhanced clinical outcomes and reduced costs when compared to Beclomethasone dipropionate/Formoterol 100/6 μg b.i.d.


Photodynamic Inactivation of Actinomyces naeslundii in Comparison With Chlorhexidine and Polyhexanide--A New Approach for Antiseptic Treatment of Medication-Related Osteonecrosis of the Jaw?
Local antimicrobial therapy is a fundamental principle in the treatment of lesions of medication-related osteonecrosis of the jaw. Antimicrobial photodynamic therapy (aPDT) as a local application for the treatment of microbial infections has become more widely used in recent years. In the mouth, the bone surface is in constant contact with saliva and thus cannot be kept sterile, making the development of strategies for disinfection even more important. Different methods currently in use include local rinses with chlorhexidine (CHX), polyhexanide (PHX), or aPDT. This study compared the efficiency of these 3 methods. The in vitro activity of 3 different agents against slowly growing Actinomyces naeslundii isolated from a patient with osteonecrosis was evaluated. PHX 0.04% solution, CHX 0.12% solution, and methylene blue (MB) based dye with a laser light of 660-nm wavelength (aPDT) were compared. The decrease in colony-forming units by each method was measured using an in vitro killing assay based on a water-exposed surface in a well plate. MB dye with laser (10 seconds) decreased the bacterial load by more than 4 orders of magnitude and was superior to PHX and CHX exposure for 60 seconds. Laser exposure alone and MB dye exposure alone decreased bacterial loads slightly, but less efficiently than 60-second exposure to PHX or CHX. The most effective means of decreasing colony-forming units was achieved by a combination of laser light and dye, which also can be used clinically.


Terrigenous organic carbon drives methane dynamics in cascade reservoirs in the upper Yangtze China.
Methane (CH4) emissions from freshwaters to the atmosphere have a profound impact on global atmospheric greenhouse gas (GHG) concentrations. Anthropogenic footprints such as dam construction and reservoir operation significantly changed the fate and transport of CH4 in freshwaters. The source of particulate organic carbon (POC) in reservoirs is a critical factor controlling CH4 production and emissions. However, little is known of how reservoir operation mediates the transport of POC and regulates CH4 accumulation in cascade hydroelectric reservoirs. Here, spatial and temporal variations in POC and CH4 were explored in the Xiluodu (XLD) and Xiangjiaba (XJB) reservoirs which are deep valley cascade reservoirs located in the main channel of the upper Yangtze River. Based on the δ13C-POC and N/C mole ratio of particulate organic matter, the results of multi-endmember stable isotope mixing models by a Bayesian model showed that terrigenous POC and autochthonous POC accounted for approximately 55% ± 18% and 43% ± 19% (SD, n = 179) of POC, respectively. Together with other hydrological and environmental parameters, we found that the input of terrigenous POC was dominantly influenced by water level variations and flow regulation due to reservoir operation. The cumulative effect of POC caused by cascade dams was not apparent. Terrigenous POC were more likely to drive CH4 accumulation in our study. Evident low level of CH4 in both reservoirs were likely affected by low sedimentation of POC and microbial CH4 oxidation. We hope our study could provide a conceptual framework for further modeling of CH4 dynamics in cascade reservoirs.


Stage II differentiated thyroid cancer: A mixed bag.
AJCC-TNM Stage II well-differentiated thyroid cancer (WDTC) comprises T2N0M0 tumors in patients ≥45 years of age or metastatic WDTC in patients younger than 45 years. The objectives of this study were to assess the oncological outcome of stage II WDTC and to compare the oncological outcome of metastatic WDTC in patient younger (stage II) and older (stage IVC) than 45 years. This study involved review of clinical presentation and oncological outcome of population cohort of 2,128 consecutive WDTC, diagnosed during 1970-2010 that includes 215 Stage II WDTC and 61 metastatic WDTC. Cox proportional hazard model was used to assess independent impact of prognostic factors on disease-specific survival (DSS) and disease-free survival (DFS) as calculated by Kaplan-Meier method. Metastatic and non-metastatic stage II WDTC had a 15-year DSS of 41.7% and 96.7%, respectively (P < 0.001). Multivariable analysis showed a 52 times higher risk of death in metastatic stage II WDTC and the DSS of metastatic stage II WDTC was not statistically different from that of stage IVC WDTC. Metastatic stage II WDTC is very different from non-metastatic stage II WDTC with oncological outcome similar to stage IVC WDTC.


Localization of transglutaminase 1 mRNA in normal and psoriatic epidermis by non-radioactive in situ hybridization.
We examined the expression of the transglutaminase 1 (TGase 1) gene in frozen sections of normal and psoriatic epidermis by means of non-radioactive in situ hybridization with digoxigenin-labelled cRNA probes. TGase 1 mRNA was expressed in the granular layer of normal epidermis, regardless of ortho- or hyperkeratosis. However, in psoriatic epidermis, TGase 1 mRNA was detected in the suprabasal spinous layer, but not in the subcorneal layer. These results indicate that TGase 1 gene expression is limited to the last stage of keratinization in normal epidermis and this regulation is disturbed in psoriasis.


Dynamic Tuning of a Thin Film Electrocatalyst by Tensile Strain.
We report the ability to tune the catalytic activities for the hydrogen evolution reaction (HER) and oxygen evolution reaction (OER) by applying mechanical stress on a highly n-type doped rutile TiO2 films. We demonstrate through operando electrochemical experiments that the low HER activity of TiO2 can reversibly approach those of the state-of-the-art non-precious metal catalysts when the TiO2 is under tensile strain. At 3% tensile strain, the HER overpotential required to generate a current density of 1 mA/cm2 shifts anodically by 260 mV to give an onset potential of 125 mV, representing a drastic reduction in the kinetic overpotential. A similar albeit smaller cathodic shift in the OER overpotential is observed when tensile strain is applied to TiO2. Results suggest that significant improvements in HER and OER activities with tensile strain are due to an increase in concentration of surface active sites and a decrease in kinetic and thermodynamics barriers along the reaction pathway(s). Our results highlight that strain applied to TiO2 by precisely controlled and incrementally increasing (i.e. dynamic) tensile stress is an effective tool for dynamically tuning the electrocatalytic properties of HER and OER electrocatalysts relative to their activities under static conditions.


Crystal structures of photosystem II from a cyanobacterium expressing psbA2 in comparison to psbA3 reveal differences in the D1 subunit.
Three psbA genes (psbA1, psbA2, and psbA3) encoding the D1 subunit of photosystem II (PSII) are present in the thermophilic cyanobacterium Thermosynechococcus elongatus and are expressed differently in response to changes in the growth environment. To clarify the functional differences of the D1 protein expressed from these psbA genes, PSII dimers from two strains, each expressing only one psbA gene (psbA2 or psbA3), were crystallized, and we analyzed their structures at resolutions comparable to previously studied PsbA1-PSII. Our results showed that the hydrogen bond between pheophytin/D1 (PheoD1) and D1-130 became stronger in PsbA2- and PsbA3-PSII due to change of Gln to Glu, which partially explains the increase in the redox potential of PheoD1 observed in PsbA3. In PsbA2, one hydrogen bond was lost in PheoD1 due to the change of D1-Y147F, which may explain the decrease in stability of PheoD1 in PsbA2. Two water molecules in the Cl-1 channel were lost in PsbA2 due to the change of D1-P173M, leading to the narrowing of the channel, which may explain the lower efficiency of the S-state transition beyond S2 in PsbA2-PSII. In PsbA3-PSII, a hydrogen bond between D1-Ser270 and a sulfoquinovosyl-diacylglycerol molecule near QB disappeared due to the change of D1-Ser270 in PsbA1 and PsbA2 to D1-Ala270. This may result in an easier exchange of bound QB with free plastoquinone, hence an enhancement of oxygen evolution in PsbA3-PSII due to its high QB exchange efficiency. These results provide a structural basis for further functional examination of the three PsbA variants.


Biomonitoring using accessible human cells for exposure and health risk assessment.
A major goal for genetic toxicologist is to provide precise information on exposure and health risk assessment for effective prevention of health problems. A frequently used approach for population study has been to utilize readily available blood cells (lymphocytes and red blood cells) as sentinel cell types to detect biological effects from exposure and to provide early warning signals for health risk. However, such approach still cannot be used reliably for developing strategies in risk assessment and disease prevention. It is possible that other available cell types which are more representative of the target cells for disease may be used to overcome the deficiency. In this report, the use of non-blood cells for biomonitoring is briefly reviewed. Their usefulness in certain exposure condition is highlighted and their effectiveness in documenting exposure compared with other cell types such as the traditional blood cells is presented. It is obvious that the decision in using these non-blood cells in biomonitoring is based on the exposure condition and the experimental design. Nevertheless, monitoring studies using non-blood cells should be encouraged with emphasis on providing dose-response information, comparative response with other cell types and effectiveness for health risk assessment.


The role of cell mixing in branchial arch development.
Compartmental structures are the basis of a number of developing systems, including parts of the vertebrate head. One of the characteristics of a series of compartments is that mixing between cells in adjacent units is restricted. This is a consequence of differential chemoaffinity between neighbouring cells in adjacent compartments. We set out to determine whether mesenchymal cells in the branchial arches and their precursors show cell-mixing properties consistent with a compartmental organisation. In chimaeric avian embryos we found no evidence of preferential association or segregation of neural crest cells when surrounded by cells derived from a different axial level. In reassociation assays using mesenchymal cells isolated from chick branchial arches at stage 18, cells reformed into clusters without exhibiting a preferential affinity for cells derived from the same branchial arch. We find no evidence for differential chemoaffinity in vivo or in vitro between mesenchymal cells in different branchial arches. Our findings suggest that branchial arch mesenchyme is not organised into a series of compartments.


Human interferon inhibits the growth of established human breast tumours in the nude mouse.
In this paper we describe a model system for looking at the effects of human interferon, IFN, on an established human tumour. Highly purified human IFN derived from lymphoblastoid cells (HuIFN alpha-Namalwa) strongly inhibited the growth of a human breast cancer growing as a xenograft in nude mice. The effect was dose-dependent, required daily treatment for an optimal effect and was time-dependent, little inhibition being seen before 2 weeks of therapy. With the doses used, however, neither tumour regression nor disappearance were seen and on morphological examination, treated tumours appeared as miniatures of control tumours. The inhibition of the tumour by HuIFN alpha-Namalwa appeared to be due to a direct effect on the human cells as this IFN had little effect on the mouse immune system in vitro as measured by NK cells activity. Also HuIFN therapy had no effect on levels of an interferon induced enzyme, 2-5A synthetase, in the mouse spleen cells but stimulated this enzyme in the human tumour.


Local anaesthetics inhibit cholinergic and non-cholinergic neural and muscular contractions in avian tracheal smooth muscle.
The effects of five different local anaesthetics: lignocaine, prilocaine, etidocaine, mepivacaine, bupivacaine, on tone, contractility, and on cholinergic and non-cholinergic responses of chick tracheal smooth muscle were studied and compared in vitro. The cholinergic and non-cholinergic contractions of the tracheal smooth muscle were elicited by electrical field stimulation, at a maximal voltage with 0.2 Hz and 0.2 ms square pulse duration, in the presence of adrenergic blockers, guanethidine, propranolol, and other drugs, e.g. indomethacin. Atropine was used to reduce the cholinergic responses due to electrical field stimulation and to applied acetylcholine (ACh). Lignocaine, in low concentrations, reduced these responses and also those produced by electrical field stimulation in the presence of atropine. In high concentration (greater than 100 times clinical concentrations), lignocaine abolished all the responses and produced a sustained contracture in the muscle. Among the local anaesthetics studied, bupivacaine and lignocaine were found to be more effective than mepivacaine, prilocaine and etidocaine, in reducing the cholinergic contractions produced by electrical field stimulation and by exogenous ACh. It was suggested that lignocaine, and other local anaesthetic drugs, may have an anti-spasmic effect on the tracheal smooth muscle, in that they inhibited the contractions induced by electrical field stimulation and by depolarizing agents.


Incidence of celiac disease autoimmunity and associations with maternal tuberculosis and pediatric Helicobacter pylori infections in 4-year-old Ethiopian children followed up in an HLA genotyped birth cohort.
The prevalence of celiac disease in the general population is mainly unknown in most of sub-Saharan African countries. The aim of this study was to determine the incidence of celiac disease autoimmunity (CDA) and its associations with latent Mycobacterium tuberculosis (LMTB) and Helicobacter pylori (HP) infections in Ethiopian children aged 4 years in an HLA genotyped cohort study. Of 1,389 recruited children between 2018 and 2022, 1,046 (75.3%) had been screened at least twice for celiac disease between the ages of 2 and 4 years using a tissue transglutaminase autoantibody (tTGA) ELISA kit. Tissue TGA-positive children were retested using radio-binding assays. CDA was defined as persistent-confirmed tTGA positivity in two consecutive samples. Associations of CDA with LMTB and HP were tested in a subpopulation of 752 children born to mothers who were previously tested for LMTB with IFN-γ and anti-HP antibodies in samples collected at a mean age of 49.3 ± 5.3 months. Screening detected 38 out of 1,046 (3.6%) IgA-tTGA-positive children. Ten (1.0%) were confirmed to be positive, with six (0.6%) children diagnosed with CDA. The incidence of CDA at 4 years of age was 1.2 per 1,000 person-years. LMTB was found in 4 of 6 (66.7%) mothers with CDA children compared with 340 of 734 (46.3%) mothers of children without CDA (p = 0.424), while HP was found in 3 of 6 (50.0%) CDA children compared with 315 of 746 (42.2%) children without CDA (p = 0.702). The incidence of CDA in Ethiopian children is lower than the pooled global incidence. Neither LMTB nor HP infections are associated with CD in Ethiopian children.


Transfer and analysis of Salmonella pdu genes in a range of Gram-negative bacteria demonstrate exogenous microcompartment expression across a variety of species.
Bacterial microcompartments (MCPs) are protein organelles that typically house toxic or volatile reaction intermediates involved in metabolic pathways. Engineering bacteria to express exogenous MCPs will allow these cells to gain useful functions involving molecule compartmentalization. We cloned a 38 kb region from the Salmonella enterica serovar Typhimurium genome containing the pdu 1,2 propanediol (1,2 PD) utilization and cob/cbi genes using the FRT-Capture strategy to clone and transfer large genomic segments. We transferred this clone to a range of Gram-negative bacteria and found the clone to be functional for 1,2 PD metabolism in a variety of species including S. Typhimurium Δpdu, Escherichia coli, Salmonella bongori, Klebsiella pneumoniae, Cronobacter sakazakii, Serratia marcescens, and different Pseudomonas species. We successfully isolated MCPs expressed from the clone from several, but not all, of these strains, and we observed this utilizing a range of different media and in the absence of protease inhibitor. We also present a mini-prep protocol that allows rapid, small-scale screening of strains for MCP production. To date, this is the first analysis of cloned, exogenous microcompartment expression across several different Gram-negative backgrounds and provides a foundation for MCP use in a variety of bacterial species using a full, intact clone.


Self-sustaining pathological processes in skin psoriasis.
Clinical findings suggest that an etiological factor of skin psoriasis (SPs) is of nervous origin. Vasoactive intestinal peptide (VIP) is the most probable candidate for such a factor since VIP is the only neurotransmitter the extracellular level of which increases during SPs exacerbation and decreases in remission. VIP released from skin nerves induces keratinocyte hyperproliferation, angiogenesis, vasodilation, and other SPs-associated cutaneous pathological processes. These can go on over a prolonged period since (1) once released, VIP induces its own further secretion; (2) VIP induces release of interleukin-6 (IL6) that evokes both its own further release and release of VIP. Thereby, a vicious circle-type mechanism perpetuating and amplifying VIP secretion can function in the focuses of psoriatic damage. The mechanism described operates still more intensively under the effects of parathyroid hormone, aldosterone, and enkephalin, the blood levels of which are elevated in patients with SPs. The above explains such features of SPs as its association with human immunodeficiency virus infection, mental stress, alcohol consumption, smoking, and dependence of SPs on skin pigmentation.


Strategy for advanced hepatocellular carcinoma based on liver function and portal vein tumor thrombosis.
Tyrosine kinase inhibitor (TKI) therapy resulted in better prognosis for patients with hepatocellular carcinoma (HCC). However, some cases with Barcelona Clinic Liver Cancer (BCLC) stage C disease still had poor prognosis. This study aimed to investigate prognosis and characteristics of patients with HCC treated with TKI based on liver function and the extent of portal vein tumor thrombosis (PVTT). Patients receiving TKI therapy (n = 345) were recruited retrospectively. Child-Pugh score and albumin-bilirubin (ALBI) score were used for assessment of liver function. The extent of PVTT was classified from Vp0 to Vp4. Radiotherapy or hepatic arterial infusion chemotherapy were carried out as additional therapy to TKI. The end-point for this analysis was overall survival (OS). A total of 291 and 54 patients received sorafenib and lenvatinib as first-line TKI therapy, respectively. The median OS of patients treated with TKI were significantly stratified by ALBI grade (grade 1, 20.1 months; grade 2a, 16.3 months; grades 2b and 3, 9.8 months; P = 0.0003). The classification of PVTT significantly stratified the prognosis of patients treated with TKI (median OS: Vp0, 18.5 months; Vp1/2, 14.4 months; Vp3/4, 5.5 months; P < 0.0001). In the ALBI 2b/3 and Vp3/4 groups, the median OS of patients treated with TKI and additional therapies was significantly longer than those treated with TKI only (9.2 months vs.. 3.6 months; P = 0.0129). Liver function and PVTT are useful for stratifying prognosis of HCC patients treated with TKI. The applicative classification could lead to appropriate therapy and better prognosis.


Cognitive function and impact on driving after SARS-COV-2 infection in a man with long-standing HIV infection: a case report.
A substantial number of individuals who experience COVID-19 infection experience prolonged physical and mental symptoms after resolution of their initial infection, and among them, many individuals experience cognitive difficulties including memory lapses and executive function difficulties, often referred to as "brain fog." The possible impact of COVID-19 infection on cognition in persons with HIV-related cognitive disorders is unknown. In this report, we describe post-COVID-19 cognitive and driving function in a 62-year-old man with HIV infection since the early 1990s.


Aptitude, achievement and competence in medicine: a latent variable path model.
To develop and test a latent variable path model of general achievement, aptitude for medicine and competence in medicine employing data from the Medical College Admission Test (MCAT), pre-medical undergraduate grade point average (UGPA) and demographic characteristics for competence in pre-clinical and measures of competence (United States Licensure Examination {USMLE} Steps 1, 2, and 3). Data were gathered on 839,710 participants from 1991 to 2000 on demographic and school variables, UGPA, MCAT subtest scores and Steps 1, 2, and 3 of the United Stated Licensure Examination (USMLE). However, subsets of the total 839,710 participants included in the database were used for various analyses and the testing of a latent variable path model (LVPA). A number of preliminary descriptive and inferential techniques were used to confirm previous hypotheses and stated relationships amongst the variables of interest to the present study. Through development and testing of a latent variable path model, three latent variables measured by UGPA (general achievement), subscales of the MCAT (aptitude for medicine), and Steps 1, 2, and 3 of the USMLE (competence in medicine) were identified which resulted in a comparative fit index = .932 of the model to a large sample (n = 20,714). In a confirmatory latent variable path model we were able to identify theoretical constructs, aptitude for medicine, general achievement, and competence in medicine and their interrelationships. These are distinct but interrelated latent variables.


Predicting Prolonged Intensive Care Unit Stay Among Patients With Sepsis-Induced Hypotension.
Sepsis risk stratification tools typically predict mortality, although stays in the intensive care unit (ICU) of 24 hours or longer may be more clinically relevant for emergency department disposition. To explore predictors of ICU stay of 24 hours or longer among infected, hypotensive emergency department patients. A secondary analysis of 2 prospective, observational studies of adult patients with severe sepsis or an infection with a systolic blood pressure less than 90 mm Hg in 3 urban, academic emergency departments was performed. Patients with hypotension and infection were included. Patients with emergency department intubation, vasopressor administration, and/or death were excluded. The primary outcome was ICU stay of 24 hours or longer or death in less than 24 hours. Multivariable logistic regression was used to predict ICU stay of 24 hours or longer. Of 233 patients, 108 (46.4%) had ICU stays of 24 hours or longer. History of heart failure (odds ratio, 3.6; 95% CI, 1.5-8.3), bicarbonate level less than 20 mEq/L (odds ratio, 2.0; 95% CI, 1.1-3.8), respiratory rate greater than 20/min (odds ratio, 2.0; 95% CI, 1.1-3.7), and creatinine level greater than 2.0 mg/dL (odds ratio, 3.6; 95% CI, 1.9-6.7) were independent predictors of ICU stay of 24 hours or longer (area under curve, 0.74). The presence of 1 of these factors predicted ICU stay of 24 hours or longer (area under curve, 0.74) with 82.4% sensitivity and 49.6% specificity. These exploratory results show that heart failure, bicarbonate level of less than 20 mEq/L, tachypnea, or creatinine level greater than 2.0 mg/dL increases the likelihood of an ICU stay of 24 hours or longer among infected, hypotensive emergency department patients.


Effects of mycotoxins on the mixed lymphocyte reaction.
Effects of toxic fungal metabolites on mixed lymphocyte reaction (MLR) using mouse splenocytes and on growth of mouse myeloma cells were examined. Among 25 toxins assayed, the IC50 values of emodin, luteoskyrin, sterlgmatocystin, deoxynivalenoi, 4-acetylnivalenol, T-2 toxin and fusaric acid for the MLR were lower than those for the cytotoxicity toward the myeloma cells, suggesting that these toxins possess suppressive activity to the cellular immune system.


Haptoglobin typing of bloodstains using horizontal discontinuous polyacrylamide gel electrophoresis.
A horizontal electrophoresis method using a discontinuous polyacrylamide gel to identify haptoglobin polymorphs is described. Improvements over the traditional vertical gel method include less sample required, easy gel preparation, and shortened run time. Additionally, vertical electrophoresis equipment, which can be technically challenging, is not needed. Caucasian, Black and Hispanic databases, each created from 100 autopsy blood samples from the New York City Medical Examiner's Office, are also presented.


Mycophenolate mofetil is safe, well tolerated, and preserves lung function in patients with connective tissue disease-related interstitial lung disease.
Interstitial lung disease (ILD) frequently complicates connective tissue diseases (CTDs). Glucocorticoids and immunomodulatory agents are regarded as mainstays of therapy for CTD-related ILD; however, apart from those studies that have evaluated certain medications for patients with scleroderma, few studies have been performed. In this study, our objectives were to examine the safety and tolerability of mycophenolate mofetil (MMF) and to determine its impact on lung function in patients with CTD-ILD. In this retrospective observational study, we analyzed patients at our center who ever received MMF for CTD-ILD. We examined the frequency and severity of side effects associated with MMF and used longitudinal data analytic methods to determine the ability of MMF to maintain lung function over time. Twenty-eight patients were treated with MMF over 35.9 patient-years. The most common underlying CTD diagnosis was scleroderma (n = 9). The most common reason for initiating MMF was an adverse effect of a prior immunomodulatory agent. Six patients had clinically significant side effects related to MMF; all resolved with dose reduction. Compared to before MMF, the mean daily prednisone dose while patients were receiving MMF was lower (10 mg/d vs 15 mg/d, p = 0.09). In addition, since starting MMF, the average percentage of predicted forced vital capacity (FVC), average percentage of predicted total lung capacity, and average percentage of predicted diffusing capacity of the lung for carbon monoxide for the cohort increased by 2.3%, 4.0%, and 2.6%, respectively. MMF appears to be safe and well tolerated in patients with CTD-ILD. Larger-scale studies are needed to further evaluate the efficacy of MMF in this patient population.


Amphotropic and VSV-G-pseudotyped retroviral vectors transduce human hematopoietic progenitor cells with similar efficiency.
One restriction of retroviral gene transfer into hematopoietic stem cells is the low level of amphotropic virus receptor. In the present study, we examined whether retroviral vectors pseudotyped with the G-protein of vesicular stomatitis virus (VSV) can overcome this restriction. Human progenitor cells purified by magnetic beads and cell sorting were transduced with an amphotropic or VSV-G-pseudotyped retroviral vector containing the truncated human nerve growth factor receptor as a marker gene. Cells were prestimulated with flt-3 ligand, stem cell factor, and interleukin-3 and transduced on fibronectin. Marker gene expression was analyzed by flow cytometry. Transduction efficiencies of amphotropic and VSV-G-pseudotyped virus for CD34+ cells did not differ significantly. Gene transfer into CD34+CD38- cells, which are enriched in more immature progenitors, was not restricted and transfer efficiencies for this subset were also similar for both pseudotypes. The addition of fibronectin improved gene transfer with the amphotropic vector considerably (5- to 19.3-fold, mean 12.6), while the effect on the VSV-G-pseudotype was far less pronounced (1- to 3.9-fold, mean 2.1, P = 0.04). In conclusion, high levels of gene transfer to human hematopoietic progenitors were achieved with an optimized transduction protocol, and transduction efficiencies could not be improved further by the use of VSV-G-pseudotypes.


Extracellular potentials of a single myelinated nerve fiber in an unbounded volume conductor.
The extracellular potentials of a single myelinated nerve fiber in an unbounded volume conductor were studied. The spatial distribution of the transmembrane potential was obtained by integrating the system of partial differential equations characterizing the electric processes in the active myelinated nerve fiber. The spatial distribution of the extracellular potentials at various radial distances in the volume conductor were calculated using the line source model. Up to a certain radial distance (500 microns) the discontinuity of the action potential propagation is reflected in the extracellular potentials, while further in the volume conductor the potentials are smooth. The effect of the fiber diameter and the internodal distance on the volume conductor potentials as well as the changes in the magnitude of the extracellular potential (in the time domain) between two adjacent nodes at various radial distances were studied. The radial decline of the peak-to-peak amplitude of the extracellular potential depends on the radial coordinate r of the field point and increases with the increase of r.


Activation of Notch1 promotes development of human CD8(+) single positive T cells in humanized mice.
Notch1 mutations are found in more than 50% of human T cell acute lymphoblastic leukemia (T-ALL) cells. However, the functions of Notch1 for human T cell development and leukemogenesis are not well understood. To examine the role of Notch1, human hematopoietic stem cells (HSCs), which had been transduced with a constitutively active form of Notch1 (ICN1), were transplanted into severely immunodeficient NOD/Shi-scid-IL2rγ(null) (NOG) mice. We found that the great majority of the ICN1-expressing hematopoietic cells in the bone marrow expressed surface markers for T cells, such as CD3, CD4, and CD8, and that this T cell development was independent of the thymus. Accordingly, phenotypically mature CD8(+) single positive (SP) T cells were observed in the spleen. Furthermore, T-ALL developed in one NOG recipient mouse out of 26 that had been secondary transferred with the T cells developed in the first NOG mice. These results indicate that Notch1 signaling in HSCs promotes CD8(+) SP T cell development, and that T cell leukemogenesis may require additional oncogenic factors other than Notch1 activation.


Nanocomposite hydrogel incorporating gold nanorods and paclitaxel-loaded chitosan micelles for combination photothermal-chemotherapy.
Development of combination photothermal-chemotherapy platform is of great interest for enhancing antitumor efficacy and inhibiting tumor recurrence, which supports selective and dose-controlled delivery of heat and anticancer drugs to tumor. Here, an injectable nanocomposite hydrogel incorporating PEGylated gold nanorods (GNRs) and paclitaxel-loaded chitosan polymeric micelles (PTX-M) is developed in pursuit of improved local tumor control. After intratumoral injection, both GNRs and PTX-M can be simultaneously delivered and immobilized in the tumor tissue by the thermo-sensitive hydrogel matrix. Exposure to the laser irradiation induces the GNR-mediated photothermal damage confined to the tumor with sparing the surrounding normal tissue. Synergistically, the co-delivered PTX-M shows prolonged tumor retention with the sustained release of anticancer drug to efficiently kill the residual tumor cells that evade the photothermal ablation due to the heterogeneous heating in the tumor region. This combination photothermal-chemotherapy presents superior effects on suppressing the tumor recurrence and prolonging the survival in the Heps-bearing mice, compared to the photothermal therapy alone.


Photoprotective activities of Lignosus rhinocerus in UV-irradiated human keratinocytes.
Lignosus rhinocerus, also known as Tiger Milk Mushroom has been used traditionally to treat a variety of human conditions, including asthma, diabetes, respiratory disease, skin allergy, and food poisoning. The reported activities of Lignosus rhinocerus extracts include anti-inflammatory, anti-oxidant, anti-asthmatic, anti-microbial, anti-cancer, neuroprotection, and immune modulation effects. However, its effect on human skin is not well documented, including human skin exposed to ultraviolet light (UV). Exposure to UV can trigger various cellular responses, including inflammation, oxidative stress, DNA damage, cell death, and cellular aging. The study aims to investigate the effects of methanolic extract prepared from cultured Lignosus rhinocerus (herein referred to as TM02 and its methanol extract as TM02-ME) on UV-irradiated human keratinocytes. Powdered stock of TM02 was dissolved and sequentially extracted with different solvents to prepare the extracts and the methanol extract was subsequently characterized based on its bio-activities on HaCaT human keratinocytes. The keratinocytes were pre-treated with the methanol extract followed by UV-irradiation. Cellular responses of the HaCaT cells such as cell viability, DNA damage, as well as gene and protein expressions that were responsive to the treatments, were characterized by using bio-assays, including reverse-transcription based PCR, Western blot, cell viability, and mitochondrial Cytochrome C release assays. TM02-ME protected HaCaT cells from UV-induced DNA damage and cell death in a dose-dependent manner. Pre-treatment of HaCaT cells with TM02-ME led to a 39% reduction of cyclobutane pyrimidine dimers (CPD) and up-regulated the gene expression of REV1 and SPINK5 in UVB-irradiated HaCaT cells when compared to the control. In addition, TM-02-ME treated HaCaT cells increased the expression of BCL-XL and BCL-2 proteins which coincided with the down-regulation of mitochondrial Cyt. C release in the UV-B irradiated HaCaT cells. The results were further supported by data that showed the stable clones of HaCaT cells stably expressed BCL-XL were resistant to UVB-induced cell death. __The results showed that TM02-ME confers photoprotective activities to UVB-irradiated HaCaT cells, leading to a reduction in DNA damage and cell death as well as up-regulated the expression of REV1 and SPINK5 which are involved in DNA repair and skin barrier function, respectively. The up-regulation of pro-survival members of the BCL-2 family by TM02-ME confers protection against UVB-induced cell death.


Efficacy of aluminum chloride in severe regorafenib-associated hand-foot skin reactions: a single-arm trial.
Regorafenib, a multikinase inhibitor, causes a high frequency of hand-foot skin reactions (HFSRs). The present study evaluated the efficacy of topical aluminum chloride, a perspiration suppressant, in reducing the severity of hand-foot skin reactions (HFSRs) caused by regorafenib. The present single-arm study included patients with metastatic colorectal cancer receiving regorafenib. Aluminum chloride ointment was applied topically one week prior to the start of regorafenib treatment, and the observation period was 12 weeks. The primary endpoint was the incidence of regorafenib-related grade 3 HFSR. Secondary endpoints were the incidence of all grades of HFSR, time to any grade of HFSR, time to improvement from grade 2 or higher to grade 1 or lower, treatment discontinuation rate, treatment interruption rate or dosage reduction due to HFSR, and incidence of adverse effects of aluminum chloride. In total 28 patients were enrolled, and 27 patients were analyzed. The incidence of grade 3 HFSR was 7.4%, meeting the primary endpoint. The incidence of all grades of HFSR was 66.7%, and the median time to the occurrence of any grade of HFSR was 15 days. No patients discontinued or reduced the regorafenib dosage because of HFSR. The most common reason for the interruption of regorafenib therapy was liver dysfunction in nine patients (33%) and HFSR in three patients (11%). No serious adverse events related to aluminum chloride were observed. Aluminum chloride ointment, a drug commonly used in routine practice to treat hyperhidrosis, is safe to use, has no serious side effects, and may be effective in reducing the occurrence of severe, regorafenib-related HFSR. ClinicalTrials.gov. identifier: jRCTs031180096, Registered on 25/01/2019.


Extradural injections and manual repositioning of spine in treatment of megalgia caused by cervical spondylotic radiculopathy.
To ascertain if it is effective to use extradural injections and manual repositioning of the spine to treat megalgia caused by cervical spondylotic radiculopathy (CSR). Patients with megalgia caused by CSR were divided into a treatment group (n = 46) treated by extradural injection and manual repositioning of the spine and a control group (n = 46) treated by a conventional method. The prevalence of cure was 58.69% and the total prevalence of effective cure in the treatment group was 97.83% and was 23.91% and 78.26%, respectively, in the control group: this difference between the two groups was significant (P < 0.05). Treatment of megalgia caused by CSR by extradural injections and manual repositioning of the spine has a good curative effect with rapid analgesia and short therapeutic course.


Differentially expressed genes during fruiting body development in the aggregative ciliate Sorogena stoianovitchae (Ciliophora: Colpodea).
Sorogena stoianovitchae is the only ciliate that undergoes fruiting body development. Previously, we demonstrated that the developmental process is divided into five distinct stages: aggregation, compact aggregation, secretion of a mucous matrix, stalk elongation, and completion of the fruiting body. When S. stoianovitchae is mildly starved, several hundreds of cells aggregate beneath the water surface, and the aggregate develops into an aerial fruiting body. Essential requirements for fruiting body development are high-cell density, a light-dark cycle, and a dark period of more than 8 consecutive hours. In addition, the initial aggregation begins during the night, and light stimulation at sunrise triggers subsequent development. To elucidate the genes involved in fruiting body development, we carried out a reverse transcription-polymerase chain reaction (RT-PCR) in cells before and after such development. Thirty-six sequences with stage-specific expression patterns were cloned and partially sequenced. BLASTX search revealed that sequences with high identity for extracellular proteins (mucin, proteophosphoglycan) or membrane proteins (surface protein, TM9SF) are likely candidates for aggregation material, mucous matrix, and stalk material. Other sequences showed similarities to proteins, such as the casein kinase related to exocytosis in Paramecium, suggesting that they are involved in exocytosis signaling pathways for fruiting body development.


Arrhythmogenic right ventricular dysplasia/cardiomyopathy: review for the clinician.
Although hypertrophic obstructive cardiomyopathy remains the most common cause of sudden cardiac death in young people, rarer causes, such as arrhythmogenic right ventricular dysplasia (ARVD), are now being increasingly recognized to lead to sudden cardiac death in the younger population. Recent advances in the understanding of the genetic inheritance, etiopathogenesis, diagnosis, and treatment options of ARVD have prompted a lot of research in this form of right ventricular cardiomyopathy. The purpose of this report is to review the etiopathogenesis, clinical manifestations, diagnosis and treatment modalities for ARVD, and recent advances in the understanding of this disease entity.


Mitochondrial involvement in tracheary element programmed cell death.
The mitochondria pathway is regarded as a central component of some types of programmed cell death (PCD) in animal cells where specific signals cause the release of cytochrome c from mitochondria to trigger a proteolytic cascade involving caspases. However, plant cells lack canonical caspases, therefore a role for the mitochondria in programmed cell death in plant cells is not obvious. Using plant cells which terminally differentiate, we provide evidence supporting the involvement of mitochondria in PCD, however the release of cytochrome c is insufficient to trigger the PCD. Prior to execution of cellular autolysis initiated by the rupture of the large central vacuole to release sequestered hydrolases, mitochondria adopt a definable morphology, the inner membrane depolarizes prior to death, and cytochrome c is released from mitochondria. However, PCD can be blocked despite translocation of cytochrome c. These results suggest a role for the mitochondria in this PCD but do not support the current animal model for a causative role of cytochrome c in triggering PCD.


Detection of Mycoplasma salivarium and Mycoplasma fermentans in synovial fluids of temporomandibular joints of patients with disorders in the joints.
Thirty-six synovial fluid samples of temporomandibular joints were obtained from 33 patients with pain and anterior disk displacement (closed lock) in the joints. DNAs were prepared from the samples and amplified by a PCR-based assay specific for Mycoplasma salivarium or Mycoplasma fermentans. Of the 36 samples, five (14%), three (8%), and 19 (53%) were positive for M. salivarium, M. fermentans and both, respectively.


Role of wild birds in the spread of clade 220.127.116.11e H5N6 highly pathogenic avian influenza virus into South Korea and Japan.
H5Nx highly pathogenic avian influenza viruses (HPAIVs) have caused transboundary epizootics in poultry and wild birds. In 2016, the H5N6 subtype of clade 22.214.171.124e HPAIVs caused multiple outbreaks in Asia, including China, Japan, Korea, and Vietnam. However, the geographical spread pattern of 18.104.22.168e H5N6 HPAIV has not been clearly identified. To better understand the emergence and transmission history of 188.8.131.52e H5N6 HPAIV, we investigated the underlying epidemiologic processes associated with this viral spread by performing a Bayesian phylogeography analysis. The results revealed that wild waterfowl played a central role in the transboundary spread of clade 18.104.22.168e H5N6 HPAIV into both endemic and non-endemic countries, causing multiple incursions of the 18.104.22.168e H5N6 HPAIV into South Korea, Japan, and Vietnam. In our analysis, Guangdong province, China was estimated to be the most probable site where 188.8.131.52e H5N6 HPAIVs emerged prior to the transboundary transmissions. Continued genomic surveillance in both wild birds and poultry would be necessary for monitoring of HPAIV incursions. In addition, enhanced biosecurity would be key to preventing the HPAIV spread by minimizing contact between domestic poultry and wild birds.


Team Approach: Improving Orthopaedic Operating Room Efficiency.
The cost of surgical care accounts for almost one-third of all health care spending in the United States. Within health care spending, the operating room (OR) is one of the largest health care costs during a perioperative episode of care. Efficiency in the OR has been associated with increased productivity, safety, and quality of care. However, multiple sources of delays can contribute to inefficiency, and improving efficiency in the OR requires a systematic approach to identify and address each issue. We report on the case of a process improvement initiative implemented in a large academic institution to improve OR efficiency in outpatient orthopaedic cases, and we discuss the lessons learned through this program. Optimizing workflow in the OR requires a multidisciplinary team approach consisting of clinician leaders with common goals and open discussion regarding the needs of each team member, including circulating nurses, surgical nurses/technologists, and anesthesiologists. Our experience highlights the importance of practical, clinician-driven changes that are supported by administrative engagement, resources for staffing and equipment, and institutional flexibility, which are required to implement systemic changes to address and improve efficiency in the OR.


An evaluation of the routine use of contrast radiography as a screening test for cervical anastomotic integrity after esophagectomy.
The use of contrast radiography as a screening test for anastomotic integrity after esophagectomy is unclear. Consecutive patients undergoing either transhiatal esophagectomy (THE) or pharyngolaryngoesophagectomy (PLE) with the cervical anastomosis performed by the same single surgeon were registered into a database. Anastomotic leak rate, modality of detection, and the result of esophagram (if performed) were recorded. A total of 132 patients underwent esophagectomy (THE, 93 patients; PLE, 39 patients). Ninety-nine patients (75%) underwent contrast radiography. The overall leak rate was 12.1% (n = 16). Nine of 16 anastomotic leaks (56.3%) were detected without contrast radiography. Seven additional patients with anastomotic leaks underwent contrast radiography, 2 (28.6%) were true positives, 4 (57.1%) were false negatives, and 1 was equivocal (14.3%). Results of contrast esophagography modified patient management in only 2 instances, whereas 4 leaks were missed by this modality. Most leaks (56.3%) were diagnosed without contrast esophagography. Contrast studies changed clinical management correctly in only 2 of 132 patients, while failing to diagnose 4 of 7 possible leaks. This study suggests that its routine use after THE or PLE is not indicated.


Coexisting renal cell carcinoma and renal artery aneurysm.
We report a rare case of a patient with a renal artery aneurysm (RAA) and an ipsilateral renal cell carcinoma (RCC). This patient was treated with simultaneous aneurysmectomy and radical nephrectomy. The indications for surgical intervention for RAA remain controversial. We present a review of the literature on the clinical presentation, classification, etiology, and management of RAA and associated RCC.


Application of image analysis for grass tillering determination.
Tillering is defined as the process of above-ground shoot production by a single plant. The number of grass tillers is one of the most important parameters in ecology and breeding studies. The number of tillers is usually determined by manually counting the separated shoots from a single plant. Unfortunately, this method is too time-consuming. In this study, a new method for counting grass tillers based on image analysis is presented. The usefulness of the method was evaluated for five grass species, Phleum pratense, Lolium perenne, Dactylis glomerata, Festuca pratensis and Bromus unioloides. The grass bunches were prepared for analysis by cutting and tip painting. The images obtained were analysed using an automatic procedure with separation of shoots and other objects based on morphological parameters. It was found that image analysis allows for very quick and accurate counting of grass tillers. However, the set of morphological parameters for object recognition must be selected individually for different grass species. This method can be recommended as a replacement for the traditional, time-consuming method in grass breeding.


Sex differences and ethnic variations in speed of automatized naming.
White American-Caucasian children from 6-12 yr showed significant sex differences on four tasks of rapid automatized naming; and in each case, girls outperformed boys. American-Chinese, native Japanese and native Korean schoolchildren did not differ from American-Caucasians in naming speed when scores for boys and girls were combined. However, in the three Asian subgroups, there were no sex differences on any naming task. It is suggested that biological as well as experiential factors contribute to behavioral sex differences; and that biological (genetic) variables associated with mating group membership modify the expression of cognitive sex differences.


A stepwise mutagenesis approach using histidine and acidic amino acid to engineer highly pH-dependent protein switches.
Antibody-based drugs can be highly toxic, because they target normal tissue as well as tumor tissue. The pH value of the extracellular microenvironments around tumor tissues is lower than that of normal tissues. Therefore, antibodies that engage in pH-dependent binding at slightly acidic pH are crucial for improving the safety of antibody-based drugs. Thus, we implemented a stepwise mutagenesis approach to engineering pH-dependent antibodies capable of selective binding in the acidic microenvironment in this study. The first step involved single-residue histidine scanning mutagenesis of the antibody's complementarity-determining regions to prescreen for pH-dependent mutants and identify ionizable sensitive hot-spot residues that could be substituted by acidic amino acids to obtain pH-dependent antibodies. The second step involved single-acidic amino acid residue substitutions of the identified residues and the assessment of pH-dependent binding. We identified six ionizable sensitive hot-spot residues using single-histidine scanning mutagenesis. Nine pH-dependent antibodies were isolated using single-acidic amino acid residue mutagenesis at the six hot-spot residue positions. Relative to wild-type anti-CEA chimera antibody, the binding selectivity of the best performing mutant was improved by approximately 32-fold according to ELISA and by tenfold according to FACS assay. The mutant had a high affinity in the pH range of 5.5-6.0. This study supports the development of pH-dependent protein switches and increases our understanding of the role of ionizable residues in protein interfaces. The stepwise mutagenesis approach is rapid, general, and robust and is expected to produce pH-sensitive protein affinity reagents for various applications.


Usefulness of speckle tracking echocardiography in hypertensive crisis and the effect of medical treatment.
The acute impact of hypertensive crisis, and changes after treatment, on left ventricular (LV) systolic and diastolic function using comprehensive echocardiography, including speckle tracking, has not been well characterized. Thirty consecutive patients admitted to the hospital from the emergency room with hypertensive crisis underwent Doppler echocardiography at baseline and after blood pressure optimization. The mean age of the patients was 54 ± 13 years, with 19 men (63%). The most common presenting symptoms included dyspnea (70%), chest pain (43%), and altered mental status (13%). Mean systolic and diastolic blood pressures at presentation were 198 ± 12 and 122 ± 12 mm Hg, decreasing to 143 ± 15 and 77 ± 12 mm Hg (p <0.001 for both) after treatment. There was no significant change in LV ejection fraction between baseline and follow-up (48 ± 18% vs 46 ± 18%, p = 0.50); however, global longitudinal LV systolic strain (-10 ± 4% to -12 ± 4%, p = 0.01) and global systolic strain rate (-1.0 ± 0.4 vs -1.4 ± 0.6 s(-1), p = 0.01) significantly improved. Mean global early diastolic strain (-7.2 ± 4.0% to -9.4 ± 2.9%, p = 0.004) and early diastolic strain rate (0.3 ± 0.2 to 0.5 ± 0.4 s(-1), p = 0.05) also improved after treatment. On multivariate analysis, the independent predictors of LV longitudinal strain at follow-up were LV ejection fraction (p <0.001), heart rate (p = 0.005), systolic blood pressure (p = 0.04), and left atrial volume index (p = 0.05). In conclusion, as opposed to LV ejection fraction, LV systolic strain and strain rate were depressed during hypertensive crisis and significantly improved after medical treatment. LV diastolic function, assessed using conventional and speckle-tracking parameters, was also depressed and significantly improved after treatment.