Testosterone levels following decreases in serum osteocalcin.
Recent preclinical studies suggest that osteoblasts are able to induce testosterone production by the testis, a process mediated by osteocalcin. Bisphosphonates substantially reduce osteocalcin levels. If osteocalcin is an important regulator of testosterone levels in adult men, it would be expected that the substantial reductions in osteocalcin induced by zoledronate would impact negatively on testosterone levels. Previously, we carried out a 2-year randomized, controlled trial of annual 4 mg zoledronate in 43 HIV-infected men. To explore the relationship between osteocalcin and testosterone further, we measured serum testosterone at baseline, 3 months, and 2 years; luteinizing hormone at 3 months and 2 years; and total osteocalcin at 2 years in 28 trial participants with available blood samples. At 2 years, total osteocalcin was 39 % lower in the zoledronate group than the placebo group (zoledronate mean 10.1 [SD 3.0] μg/L, placebo 16.5 [SD 4.9] μg/L, P = 0.003). Despite these substantial differences in osteocalcin levels, testosterone levels did not change over time in either group and there were no between-group differences over time, P = 0.4 (mean change at 2 years [adjusted for baseline levels] in zoledronate group -0.4 nmol/L, 95 % CI -2.5 to 1.6; placebo group 0.4 nmol/L, 95 % CI -1.6 to 2.5). Luteinizing hormone was within the normal range and did not differ between the groups at either 3 months or 2 years. Thus, the absence of a change in testosterone despite a substantial reduction in osteocalcin following zoledronate treatment argues against a biologically significant role for osteocalcin in the regulation of testosterone in adult men. This provides reassurance that men receiving potent antiresorptive drugs are not at risk of iatrogenic hypogonadism.


Natural use of heterothermy by a small, tree-roosting bat during summer.
Little is known about the use of heterothermy by wild bats during summer, especially for tree-roosting species. Because thermal conditions within tree roosts can fluctuate widely with ambient temperature, which affects thermoregulatory energy expenditure during diurnal roosting, we measured skin temperatures of free-ranging male Nyctophilus geoffroyi (8 g) to quantify the relation between summer torpor use and roost thermal conditions. Bats roosted under bark on the northern (sunny) side of trees and entered torpor every day, usually near sunrise. Bats exhibited two bouts of torpor on most days: the first occurred in the morning, was terminated by partially passive rewarming, and was followed by a period of normothermy during the warmest part of the day; a second torpor bout occurred in the late afternoon, with arousal near sunset. On the warmest days, bats had only a single, short morning bout. On the coolest days, bats remained torpid throughout the day, and one 2-d bout was observed. Thus, presumably owing to their poorly insulated roosts and the high energetic cost of normothermy at temperatures below 30 degrees C, the extent and timing of heterothermy was closely related to the cycle of diurnal temperatures. Our study indicates that torpor use is important for energy maintenance during summer diurnal roosting of N. geoffroyi and likely of other small, tree-roosting bats.


Determining Hospital Volume Threshold for Safety of Minimally Invasive Pancreaticoduodenectomy: A Contemporary Cutpoint Analysis.
Guidelines recommend limiting minimally invasive pancreaticoduodenectomy (MIPD) to high-volume centers. However, the definition of high-volume care remains unclear. We aimed to objectively define a minimum number of MIPD performed annually per hospital associated with improved outcomes in a contemporary patient cohort. Resectable pancreatic adenocarcinoma patients undergoing MIPD were included from the National Cancer Database (2010-2017). Multivariable modeling with restricted cubic splines was employed to identify an MIPD annual hospital volume threshold associated with lower 90-day mortality. Outcomes were compared between patients treated at low-volume (≤ model-identified cutoff) and high-volume (> cutoff) centers. Among 3079 patients, 141 (5%) died within 90 days. Median hospital volume was 6 (range 1-73) cases/year. After adjustment, increasing hospital volume was associated with decreasing 90-day mortality for up to 19 (95% CI 16-25) cases/year, indicating a threshold of 20 cases/year. Most cases (82%) were done at low-volume (< 20 cases/year) centers. With adjustment, MIPD at low-volume centers was associated with increased 90-day mortality (OR 2.7; p = 0.002). Length of stay, positive surgical margins, 30-day readmission, and overall survival were similar. On analysis of the most recent two years (n = 1031), patients at low-volume centers (78.2%) were younger and had less advanced tumors but had longer length of stay (8 versus 7 days; p < 0.001) and increased 90-day mortality (7% versus 2%; p = 0.009). The cutpoint analysis identified a threshold of at least 20 MIPD cases/year associated with lower postoperative mortality. This threshold should inform national guidelines and institution-level protocols aimed at facilitating the safe implementation of this complex procedure.


Sinus node electrogram in patients with the hypersensitive carotid sinus syndrome.
Sinus node electrograms were obtained in two patients with unexplained syncope and the cardioinhibitory form of the hypersensitive carotid sinus syndrome. Direct recordings of sinus node potentials were obtained using a transvenous electrode catheter. Sinus node function was normal in both patients during standard electrophysiologic evaluation. Carotid sinus massage was performed in both patients and the sinus node electrogram was continuously recorded. After the onset of carotid sinus massage, prolongation of sinoatrial time, slowing of sinus rate of depolarization, sinoatrial exit block and finally sinus node arrest were recorded. After termination of carotid sinus massage, sinus node potentials did not precede the first atrial impulse; subsequent beats showed markedly prolonged sinoatrial times as well as changes in the P wave on the surface electrocardiogram. Sinus rate and sinoatrial time returned to control values gradually, as did the P wave configuration. Intravenous atropine (1.0 mg) abolished the abnormal response to carotid sinus massage. It is concluded that the application of carotid sinus massage in patients with the hypersensitive carotid sinus syndrome produces profound changes in sinoatrial conduction including sinoatrial exit block, as well as shifts in primary pacemaker site and sinus node arrest. These alterations in conduction and automaticity are reversible with atropine and may be secondary to denervation sensitivity to acetylcholine.


The C terminus of the large tegument protein pUL36 contains multiple capsid binding sites that function differently during assembly and cell entry of herpes simplex virus.
The largest tegument protein of herpes simplex virus type 1 (HSV1), pUL36, is a multivalent cross-linker between the viral capsids and the tegument and associated membrane proteins during assembly that upon subsequent cell entry releases the incoming capsids from the outer tegument and viral envelope. Here we show that pUL36 was recruited to cytosolic progeny capsids that later colocalized with membrane proteins of herpes simplex virus type 1 (HSV1) and the trans-Golgi network. During cell entry, pUL36 dissociated from viral membrane proteins but remained associated with cytosolic capsids until arrival at the nucleus. HSV1 UL36 mutants lacking C-terminal portions of increasing size expressed truncated pUL36 but could not form plaques. Cytosolic capsids of mutants lacking the C-terminal 735 of the 3,164 amino acid residues accumulated in the cytosol but did not recruit pUL36 or associate with membranes. In contrast, pUL36 lacking only the 167 C-terminal residues bound to cytosolic capsids and subsequently colocalized with viral and host membrane proteins. Progeny virions fused with neighboring cells, but incoming capsids did not retain pUL36, nor could they target the nucleus or initiate HSV1 gene expression. Our data suggest that residues 2430 to 2893 of HSV1 pUL36, containing one binding site for the capsid protein pUL25, are sufficient to recruit pUL36 onto cytosolic capsids during assembly for secondary envelopment, whereas the 167 residues of the very C terminus with the second pUL25 binding site are crucial to maintain pUL36 on incoming capsids during cell entry. Capsids lacking pUL36 are targeted neither to membranes for virus assembly nor to nuclear pores for genome uncoating.


Four-day continuous infusion of cisplatin and 5-fluorouracil in head and neck cancer.
A combination of cisplatin and 5-fluorouracil, both administered 4 days continuously as infusion, was assessed in advanced head and neck cancer. Of the 37 patients studied, there were 15 complete and 17 partial responses (40.5% and 45.9%, respectively). Survival is 79.1% at 22 months. None of the patients in complete response has relapsed. In general toxic effects were moderate. Given as initial treatment, the regimen is effective and of considerable use in this type of patient.


Michaelis-Menten kinetic modeling of hemoglobin A1c status facilitates personalized glycemic control.
Discrepancy between measured HbA1c and HbA1c calculated from plasma glucose is associated with higher risk for diabetic complications. However, quantification of this difference is inaccurate due to the imperfect linear conversion models. We propose to introduce a mathematical formula that correlates with the observational data and supports individualized glycemic control. We analysed 175,437 simultaneous plasma glucose and HbA1c records stored in our laboratory database. Employing the Michaelis-Menten (MM) equation, we compared the calculated HbA1c levels to the measured HbA1c levels. Data from patients with multiple records were used to establish the patients' glycemic status and to assess the predictive power of our MM model. HbA1c levels calculated with the MM equation closely matched the population's average HbA1c levels. The Michaelis constant (Km) had a negative correlation with HbA1c (r2 = 0.403). Using personalized Km values in the MM equation, 85.1% of HbA1c predictions were within 20% error (ADAG calculation: 78.4%). MM prediction also performed better in predicting pathologic HbA1c levels (0.904 AUC vs. 0.849 AUC for ADAG). MM equation is an improvement over linear models and could be readily employed in routine diabetes management. Km is a reliable and quantifiable marker to characterize variations in glucose tolerance.


The Pediatric American Pain Society Patient Outcomes Questionnaire (Pediatric APS-POQ): Development and Initial Psychometric Evaluation of a Brief and Comprehensive Measure of Pain and Pain Outcomes in Hospitalized Youth.
Pediatric pain assessment in the hospital traditionally involves the patient's self-report of pain intensity using a numeric rating scale, which does not capture the complexity of the pain experience. No valid, comprehensive measure of pain in hospitalized youth exists. This study was designed to develop and conduct initial psychometric testing of the Pediatric American Pain Society Patient Outcomes Questionnaire (Pediatric APS-POQ), a comprehensive patient-reported measure of pain and pain outcomes in hospitalized youth. A multidisciplinary group of pediatric pain researchers and clinicians collaborated to adapt the adult APS-POQ Revised to pediatrics, including a patient-report and parent proxy version. The adapted measures were administered to 218 pediatric inpatients (age M = 13.4 years, 56% female) and 214 of their parents (80% mothers) at 4 US children's hospitals. The measure was feasible to administer within the inpatient setting and was acceptable and understandable to pediatric patients and their parents. Internal consistency was adequate for both patient-report and parent proxy (α = 0.77). Confirmatory factor analysis supported the following 6 domains, consistent with the adult measure: pain intensity, functional interference, emotional response, side effects, perceptions of care, and usual pain. Additional research is needed to further support the reliability and validity of this measure in diverse clinical populations. PERSPECTIVE: To reduce the impact of pain on hospitalized youth, pediatric pain assessment must move beyond ratings of pain intensity. The Pediatric APS-POQ provides a brief but comprehensive assessment of pain and pain outcomes in hospitalized children and adolescents, which will allow for greater individualization in hospital-based pain management and quality improvement purposes.


Short-Term Supplemental Dietary Potassium from Potato and Potassium Gluconate: Effect on Calcium Retention and Urinary pH in Pre-Hypertensive-to-Hypertensive Adults.
Potassium supplementation has been associated with reduced urinary calcium (Ca) excretion and increased Ca balance. Dietary interventions assessing the impact of potassium on bone are lacking. In this secondary analysis of a study designed primarily to determine blood pressure effects, we assessed the effects of potassium intake from potato sources and a potassium supplement on urinary Ca, urine pH, and Ca balance. Thirty men (n = 15) and women (n = 15) with a mean ± SD age and BMI of 48.2 ± 15 years and 31.4 ± 6.1 kg/m2, respectively, were enrolled in a cross-over, randomized control feeding trial. Participants were assigned to a random order of four 16-day dietary potassium interventions including a basal diet (control) of 2300 mg/day (~60 mmol/day) of potassium, and three phases of an additional 1000 mg/day (3300 mg/day(~85 mmol/day) total) of potassium in the form of potatoes (baked, boiled, or pan-heated), French fries (FF), or a potassium (K)-gluconate supplement. Calcium intake for all diets was approximately 700-800 mg/day. Using a mixed model ANOVA there was a significantly lower urinary Ca excretion in the K-gluconate phase (96 ± 10 mg/day) compared to the control (115 ± 10 mg/day; p = 0.027) and potato (114 ± 10 mg/day; p = 0.033). In addition, there was a significant difference in urinary pH between the supplement and control phases (6.54 ± 0.16 vs. 6.08 ± 0.18; p = 0.0036). There were no significant differences in Ca retention. An increased potassium intake via K-gluconate supplementation may favorably influence urinary Ca excretion and urine pH. This trial was registered at ClinicalTrials.gov as NCT02697708.


Stuttering severity and treatment outcome.
This study evaluated a multidimensional treatment of stuttering (awareness training, regulated breathing, cognitive restructuring and relapse prevention) for mild, moderate and severe stutterers. Behavioral measures (percentage of syllables stuttered and rate of speech) and cognitive measures (self-efficacy perception, locus of control and Erickson Scale) were taken. Using three multiple baseline designs across subjects, results showed that all mild stutterers and most moderate stutterers were clinically improved at the end of treatment and at six months follow-up. No severe stutterers (more than 15% of syllables stuttered) achieved clinical improvement. Self-efficacy perceptions were quite accurate in predicting the performance of successfully treated patients. No consistent changes were observed on the locus of control or the Erickson Scale. The clinical and theoretical implications of the results are discussed.


The experimental studies on the determination of the ground and excited state dipole moments of some hemicyanine dyes.
The ground state (mu(g)) and excited state (mu(e)) dipole moments of 15 hemicyanine dyes were studied at room temperature. They were estimated from solvatochromic shifts of the absorption and the fluorescence spectra as function of the solvent dielectric constant (epsilon) and refractive index (n). In this paper we applied the Stokes shift phenomena not only for the determination of the difference in the dipole moment of excited state and ground state, but to determine the value of polarizability alpha as well. The obtained results show that excited state dipole moments of hemicyanine dyes are in the range from 5 to 15 Debye, and the difference between the excited and ground state dipole moments vary from 1 to 7 Debye. The excited and ground state dipole moments difference (mu(e)-mu(g)) obtained for selected dyes are positive. It means that the excited states of the dyes under the study are more polar than the ground state ones. Additionally, the value of both polarizability (alpha) and the Onsager radius (a) of each investigated hemicyanine dye molecule are determined, and the ratio of alpha/a(3) for each dye were calculated, which oscillate from 0.29 to 5.21. The increase in dipole moment has been explained in terms of the nature of excited state and its resonance structure.


A prospective, randomized, controlled trial of the efficacy and safety of fibrin pad as an adjunct to control soft tissue bleeding during abdominal, retroperitoneal, pelvic, and thoracic surgery.
This study evaluated the hemostatic effectiveness and safety of Fibrin Pad (Omrix Biopharmaceuticals Ltd.) vs absorbable hemostat in patients undergoing nonemergent surgery. Fibrin Pad is a topical absorbable hemostat designed to be effective in a variety of soft tissues and across multiple bleeding intensities. Patients 18 years and older, requiring abdominal, retroperitoneal, pelvic, or thoracic (noncardiac) surgery and with an appropriate soft-tissue target bleeding site (TBS), were randomized to receive Fibrin Pad or absorbable hemostat (NCT00658723). Patients were stratified by bleeding severity at the TBS. Assessments included percentage of patients achieving hemostasis at 4 minutes after randomization with no rebleeding requiring treatment during the subsequent 6 minutes (primary endpoint), proportion of patients achieving hemostasis at 10 minutes, and incidence of treatment failure. On the primary endpoint, 98.3% of patients with Fibrin Pad and 53.3% with absorbable hemostat achieved hemostasis at 4 minutes (p < 0.0001). The treatment differential was magnified (efficacy was maintained with Fibrin Pad but decreased with absorbable hemostat) with increasing bleeding intensity: in patients with mild bleeding, 100.0% vs 80.0% achieved hemostasis with Fibrin Pad and absorbable hemostat (p = 0.03), respectively; rates were 96.6% vs 26.7%, respectively (p < 0.0001) with moderate bleeding. Percentages of patients who achieved hemostasis at 10 minutes were: Fibrin Pad, 98.3% and absorbable hemostat, 73.3% (p < 0.0001). Incidences of adverse events were comparable between groups. Fibrin Pad is superior to absorbable hemostat (SURGICEL Original Absorbable Hemostat [Ethicon]) in soft-tissue bleeding control and is safe and effective as an adjunct for rapidly and reliably achieving hemostasis for soft-tissue bleeding during surgery.


Prognostic value of nocturnal blood pressure and reverse-dipping status on the occurrence of cardiovascular events in hypertensive diabetic patients.
To assess whether reverse-dipping status is associated with cardiovascular (CV) events such as CV death, myocardial infarction (MI) or stroke in diabetic patients with hypertension. A total of 97 diabetic patients underwent their first ambulatory blood pressure monitoring (ABPM 1). "Reverse dippers" were defined as patients with a nighttime systolic and/or diastolic blood pressure (BP) greater than daytime systolic and/or diastolic BP. Other patients were called "others". A second ABPM (ABPM 2) was done after a median delay of 2.6 years. Patients were then followed for a further 2.9-year median period (total median follow-up: 5.5 years). After ABPM 1, CV events occurred in 53% of the reverse dippers (n=15) and in 29% of the others (n=82). Kaplan-Meier curves showed significant differences between the two groups (P=0.003). Mean nighttime systolic BP on ABPM 1 was 148+/-23 mmHg and 142+/-19 mmHg in patients who did and did not experience a CV event, respectively. With Cox analysis adjusted for confounders, a 10 mmHg increase in nighttime systolic BP was associated with a 35% increase in the risk of a CV event (hazard ratio [HR]: 1.35, P=0.003). The HR for a CV event in reverse- versus nonreverse-dipping status was 2.79 (P=0.023). After ABPM 2, the relationship between the reverse-dipping status and occurrence of CV events was no longer evident (P=0.678). Nighttime systolic BP remained predictive of CV events (P=0.001). These findings suggest that nighttime systolic BP per se appeared to be a stronger predictor of an excess risk of CV events compared with reverse-dipping status.


What works to prevent falls in older adults dwelling in long term care facilities and hospitals? An umbrella review of meta-analyses of randomised controlled trials.
Preventing falls in long term care facilities (LTCF) and hospitals is an international priority. Many interventions have been investigated and summarised in meta-analyses (MA) and there is a need to synthesise the top of the hierarchy of evidence in one place. Therefore we conducted an umbrella review of MA of randomised controlled trials (RCTs) of falls prevention interventions LTCF and hospitals. Two independent reviewers searched major electronic databases from inception till October 2014 for MA containing ≥3 RCTs investigating any intervention to prevent falls in LTCF or hospitals in older adults aged ≥60 years. Methodological quality was assessed by the AMSTAR tool and data were narratively synthesised. The methodological quality of the MA was moderate to high across the 10 included MA. Nine MA provided data for LTCF and only two considered hospital settings. Only one MA defined a fall and two reported adverse events (although minor). Consistent evidence suggests that multifactorial interventions reduce falls (including the rate, risk and odds of falling) in LTCF and hospitals. Inconsistent evidence exists for exercise and vitamin D as single interventions in LTCF, whilst no MA has investigated this in hospitals. No evidence exists for hip protectors and medication review on falls in LTCF. In conclusion, multifactorial interventions appear to be the most effective interventions to prevent falls in LTCF and hospital settings. This is not without limitations and more high quality RCTs are needed in hospital settings in particular. Future RCTs and MA should clearly report adverse events.


Dentists' Self-evaluated Ability in Diagnosing and Updating About Pulpotomy.
This study aimed to self-evaluate the knowledge of different dental professionals' profiles in Brazil on diagnosing and indicating pulpotomy in primary teeth and how these groups updated on this theme. Three groups (G1: professors, G2: specialists, and G3: professionals) answered an online questionnaire containing 20 questions and returned 416 questionnaires. Data were subjected to Chi-square associative tests and t tests, with P < .05. G1 and G2 reported to indicate pulpotomy, highlighting the tendency of these groups towards seeking updates on this subject. All 3 groups used academic materials for updating. However, G2 had a high demand for updates through congresses, whilst G3 had a high demand for updates through social media (Google and Google Scholar). The most cited indication by G1 and G2 was "accidental pulp exposure" and by G3 was "teeth with extensive carious lesions that have involved the pulp with radiographic confirmation." All 3 groups opted for pulpotomy to keep the tooth in the arch. Thus, G1 and G2 are the groups who are more updated on the subject, using mainly academic materials, which can be associated with the fact that such groups indicated pulpotomy in primary teeth. The lower interest in searching about pulpotomy in G3 can be understood as a nonclinical application of the topic for this group. Professionals who work directly with paediatric dentistry (professors or specialists) felt more capable of diagnosing and treating cases of pulpotomy. In addition, most of the professionals used scientifically based sources for getting information and actualisation on the subject of pulp therapies.


Management of clinically negative neck for the patients with head and neck squamous cell carcinomas in the modern era.
Management of the cervical metastases is of paramount importance in the treatment of patients with head and neck squamous cell carcinoma (HNSCC). Head and neck oncologists continue to debate the appropriate approach of the clinically negative neck among patients with HNSCC. There are three management options: (A) Observation, reserving therapeutic neck dissection for only those patients who subsequently develop metastatic disease in the neck. (B) Staging with reserving definitive treatment for those who are found to have subclinical disease in the neck. Staging may require the use of imaging techniques or "staging" neck dissection. In the latter case a selective neck dissection (SND) is usually recommended, but it is still controversial if this surgical procedure for clinically negative neck is a staging or a therapeutic approach. (C) Elective treatment of the neck using neck dissection, radiation therapy or both. All these strategies may be appropriate, depending on the clinical circumstances and will be discussed in this review.


Relevance of Fusion Genes in Pediatric Cancers: Toward Precision Medicine.
Pediatric cancers differ from adult tumors, especially by their very low mutational rate. Therefore, their etiology could be explained in part by other oncogenic mechanisms such as chromosomal rearrangements, supporting the possible implication of fusion genes in the development of pediatric cancers. Fusion genes result from chromosomal rearrangements leading to the juxtaposition of two genes. Consequently, an abnormal activation of one or both genes is observed. The detection of fusion genes has generated great interest in basic cancer research and in the clinical setting, since these genes can lead to better comprehension of the biological mechanisms of tumorigenesis and they can also be used as therapeutic targets and diagnostic or prognostic biomarkers. In this review, we discuss the molecular mechanisms of fusion genes and their particularities in pediatric cancers, as well as their relevance in murine models and in the clinical setting. We also point out the difficulties encountered in the discovery of fusion genes. Finally, we discuss future perspectives and priorities for finding new innovative therapies in childhood cancer.


Activation of recombinant h 5-HT1B and h 5-HT1D receptors stably expressed in C6 glioma cells produces increases in Ca2+-dependent K+ current.
The putative coupling between stably expressed recombinant h 5-HT1B or h 5-HT1D receptors and K+ channels which regulate excitability was investigated in C6 glioma cells. Outward K+ currents (IK) were examined in nontransfected C6 glioma cells and in cells expressing cloned h 5-HT1B or h 5-HT1D receptors using the patch-clamp technique in the whole-cell configuration. IK was elicited by a depolarizing step from a holding potential of -60 mV. In C6 glioma cells expressing either recombinant h 5-HT1B or h 5-HT1D receptors, sumatriptan similarly increased IK in a concentration-dependent manner (maximum increase 19.4+/-7.2%, n=8, P<0.05 and 25.1+/-3.9%, n=6, P<0.001, respectively) with EC50 values (geometric mean with 95% confidence intervals in parentheses) of 56.3 nM (7.9-140 nM) and 68.7 nM (16-120 nM), respectively. Sumatriptan failed to elicit increases in IK in non-transfected cells, confirming a specific involvement of the respective membrane h 5-HT1B and h 5-HT1D receptors in transfected C6 cells. In the presence of the mixed 5-HT1B/D receptor antagonist GR 127935 (0.1 microM), sumatriptan (1 microM) failed to significantly increase IK in C6 cells expressing h 5-HT1B receptors (-7.5+/-3.5%, P=NS, n=6), although a higher concentration of GR 127935 (1 microM) was required to significantly inhibit sumatriptan-evoked increases in IK in C6 cells expressing h 5-HT1D receptors (-1.8+/-3.5%, P=NS, n=6), confirming that sumatriptan-evoked responses were indeed mediated by h 5-HT1B and h 5-HT1D receptors, respectively. In C6 cells expressing either cloned h 5-HT1B or h 5-HT1D receptors, sumatriptan-induced increases in IK were prevented by the calcium chelator EGTA (5 mM) when included in the patch pipette (maximum increase 0.57+/-0.6%, n=3, P=NS and -2.8+/-1.6%, n=5, P=NS, respectively). In C6 cells expressing cloned h 5-HT1B receptors, sumatriptan (1 microM) similarly failed to significantly increase IK in the presence of dibutyryl cAMP (10 microM) or when a nominally Ca2+-free medium was included in the patch pipette (-19.4+/-5.1%, n=5 and -5.2+/-4.3%, n=5, respectively, P=NS in each case). In addition, the Ca2+-dependent K+ channel blockers iberiotoxin (0.1 microM) and tetraethylammonium (TEA, 1 mM) abolished sumatriptan-induced increases in IK (-0.5+/-1.0%, n=4 and -3.9+/-3.1%, n=4, respectively, P=NS in each case) in C6 cells expressing h 5-HT1B receptors, confirming the involvement of Ca2+-dependent K+ channels. In C6 cells expressing cloned h 5-HT1B receptors, sumatriptan (1 microM) similarly failed to significantly increase IK after 30-min incubation with thapsigargin (1 microM) or when heparin (2 mg/ml) was included in the patch pipette (1.1+/-0.4%, n=5 and 1.2+/-2.4%, n=5, respectively, P=NS). In conclusion, evidence is provided that both recombinant h 5-HT1B and h 5-HT1D receptors stably transfected in C6 glioma cells are positively coupled to Ca2+-dependent K+ channels, and the outward hyperpolarizing current mediated by these channels is dependent upon IP3 receptor-mediated intracellular Ca2+ release.


Linkage between lipid droplet formation and nuclear deformation in HeLa human cervical cancer cells.
Because lipid droplets (LDs) and the nucleus are cellular organelles that regulate seemingly very different biochemical processes, very little attention has been focused on their possible interplay. Here, we report a correlation between nuclear morphology and cytoplasmic LD formation in HeLa human cervical cells. When the cells were treated with oleic acid (OA), LDs were formed in the cytoplasm, but not in the nucleoplasm. Interestingly, cells harboring OA-induced cytoplasmic LDs showed deformity of the nucleus, particularly at the nuclear rim. Conversely, when alteration from a single spherical nuclear shape to a multinucleated form was enforced by coadministration of paclitaxel and reversine, a significant amount of LDs was detected in the cytoplasm of the multinucleated cells. These two distinct pharmacological culture conditions not only allow analysis of the previously underappreciated organelle relationship, but also provide insights into the mutual affectability of LD formation and nuclear deformation.


[CYP2E1 gene polymorphism and ovarian cancer risk in the Yakut population].
The CYP2E1 gene polymorphism has been studied in Yakut women with ovarian cancer and without cancer. The two groups have been found to substantially differ in the frequency of the CYP2E1* 1D allele (with a 96-bp insertion in the promoter region of the gene): it is more frequent in healthy women (16.3 versus 7.4%, P = 0.007).


Oil-material fractionation in Gulf deep water horizontal intrusion layer: Field data analysis with chemodynamic fate model for Macondo 252 oil spill.
Among the discoveries of the Deepwater Horizon blowout was the so-called "sub-surface plume"; herein termed the "oil-trapping layer". Hydrocarbons were found positioned at ~1100-1300m with thickness ~100-150m and moving horizontally to the SW in a vertically stratified layer at the junction of the cold abyssal water and the permanent thermocline. This study focuses on its formation process and fate of the hydrocarbons within. The originality of this work to the field is two-fold, first it provides a conceptual framework which places layer origin in the context of a horizontal "intrusion" from the near-field, vertical, blow-out plume and second, it offers a theoretical model for the hydrocarbon chemicals within the horizontal layer as it moves far-afield. The model quantifies the oil-material fractionation process for the soluble and fine particle. The classical Box model, retrofitted with an internal gradient, the "G-Box", allows an approach that includes turbulent eddy diffusion coupled with droplet rise velocity and reactive decay to produce a simple, explicit, transparent, algebraic model with few parameters for the fate of the individual fractions. Computations show the soluble and smallest liquid droplets moving very slowly vertically through the layer appearing within the trapping layer at low concentration with high persistence. The larger droplets move-through this trapping zone quickly, attain high concentrations, and eventually form the sea surface slick. It impacts the field of oil spill engineering science by providing the conceptual idea and the algorithms for projecting the quantities and fractions of oil-material in a deep water, horizontal marine current being dispersed and moving far afield. In the field of oil spill modeling this work extends the current generation near-field plume source models to the far-field. The theory portrays the layer as an efficient oil-material trap. The model-forecasted concentration profiles for alkanes and aromatics against the available field data support the proposed theory and the resulting model.


Pollution mitigation and carbon sequestration by an urban forest.
At the beginning of the 1900s, the Canberra plain was largely treeless. Graziers had carried out extensive clearing of the original trees since the 1820s leaving only scattered remnants and some plantings near homesteads. With the selection of Canberra as the site for the new capital of Australia, extensive tree plantings began in 1911. These trees have delivered a number of benefits, including aesthetic values and the amelioration of climatic extremes. Recently, however, it was considered that the benefits might extend to pollution mitigation and the sequestration of carbon. This paper outlines a case study of the value of the Canberra urban forest with particular reference to pollution mitigation. This study uses a tree inventory, modelling and decision support system developed to collect and use data about trees for tree asset management. The decision support system (DISMUT) was developed to assist in the management of about 400,000 trees planted in Canberra. The size of trees during the 5-year Kyoto Commitment Period was estimated using DISMUT and multiplied by estimates of value per square meter of canopy derived from available literature. The planted trees are estimated to have a combined energy reduction, pollution mitigation and carbon sequestration value of US$20-67 million during the period 2008-2012.


YM155 induces caspase-8 dependent apoptosis through downregulation of survivin and Mcl-1 in human leukemia cells.
Survivin, a member of the inhibitor of apoptosis protein (IAP) family, is highly expressed in various kinds of tumors. In the present study, we investigated the cytotoxic mechanism of YM155, a unique small-molecule inhibitor of survivin, in human myelogenous leukemia cells. YM155 potently inhibited the cell growth of HL-60 and U937 cells with the half-maximal inhibitory concentration (IC50) value of 0.3 nM and 0.8 nM, respectively. YM155 significantly suppressed the levels of mRNA expression and protein of survivin in HL-60 and U937 cells. In addition, we also found that YM155 down-regulated the level of Mcl-1, another critical anti-apoptotic protein, in both HL-60 and U937 cells. Treatment of HL-60 and U937 cells with YM155 induced apoptosis concomitant with the activation of caspase-8 and caspase-3. Interestingly, we have found that caspase-8 inhibitor Z-IETD-FMK strongly inhibited YM155-induced apoptosis in HL-60 and U937 cells. When cells were pretreated with Z-IETD-FMK, the activation of caspase-3 was completely abolished, suggesting that caspase-8 may be involved in the activation of caspase-3 during YM155-induced apoptosis. We demonstrated for the first time that YM155 induces caspase-8 dependent apoptosis through downregulation of survivin and Mcl-1 in human leukemia cells.


Management of children with co-occurring sleep disordered breathing and hearing loss.
To examine hearing loss incidence in a pediatric otolaryngology patients with sleep disordered breathing, and secondarily, identify possible disparities in management. A retrospective case-control study of pediatric patients with sleep-disordered breathing or obstructive sleep apnea from 2012 to 2019 was conducted at a private, not-for-profit, academic, tertiary care center. Study parameters were extracted from the electronic medical record and compared in study groups of patients with sleep-disordered breathing generated based on presence of hearing loss (cases) versus absence (controls). Study parameters were also compared in study groups based on timing of hearing loss onset. 14.8% of patients with sleep disordered breathing had hearing loss. Hearing loss was associated with an increased risk of undergoing adenotonsillectomy (OR 1.632 [1.294-2.058], p < 0.001, adjusted for age). In patients who underwent polysomnographic testing, 9.8% patients had pre-existing hearing loss and 12% patients developed hearing loss over the study period. Patients with pre-existing hearing loss and those who developed hearing loss had significantly more visits with otolaryngology compared to controls (p < 0.001). Hearing loss did not significantly delay adenotonsillectomy. More patients who developed hearing loss had adenotonsillectomies (OR 2.475 [1.672-3.663], p < 0.001, adjusted for age) versus controls. This difference was not identified in patients with pre-existing hearing loss. Patients with evidence of hearing loss in addition to sleep disordered breathing had more adenotonsillectomies performed and more clinic visits. Further work must be done to understand the associations and implications of hearing loss in this population.


Metabolic effects of p-coumaric acid in the perfused rat liver.
The p-coumaric acid, a phenolic acid, occurs in several plant species and, consequently, in many foods and beverages of vegetable origin. Its antioxidant activity is well documented, but there is also a single report about an inhibitory action on the monocarboxylate carrier, which operates in the plasma and mitochondrial membranes. The latter observation suggests that p-coumaric acid could be able to inhibit gluconeogenesis and related parameters. The present investigation was planned to test this hypothesis in the isolated and hemoglobin-free perfused rat liver. Transformation of lactate and alanine into glucose (gluconeogenesis) in the liver was inhibited by p-coumaric acid (IC50 values of 92.5 and 75.6 microM, respectively). Transformation of fructose into glucose was inhibited to a considerably lower degree (maximally 28%). The oxygen uptake increase accompanying gluconeogenesis from lactate was also inhibited. Pyruvate carboxylation in isolated intact mitochondria was inhibited (IC50 = 160.1 microM); no such effect was observed in freeze-thawing disrupted mitochondria. Glucose 6-phosphatase and fructose 1,6-bisphosphatase were not inhibited. In isolated intact mitochondria, p-coumaric acid inhibited respiration dependent on pyruvate oxidation but was ineffective on respiration driven by succinate and beta-hydroxybutyrate. It can be concluded that inhibition of pyruvate transport into the mitochondria is the most prominent primary effect of p-coumaric acid and also the main cause for gluconeogenesis inhibition. The existence of additional actions of p-coumaric acid, such as enzyme inhibitions and interference with regulatory mechanisms, cannot be excluded.


Impact on gait biomechanics of using an active variable impedance prosthetic knee.
An above knee amputation can have a significant impact on gait, with substantial deviations in inter-leg symmetry, step length, hip exertion and upper body involvement even when using a current clinical standard of care prosthesis. These differences can produce gait that is less efficient and less comfortable, resulting in slower and shorter distance walking, particularly with long term use. A robotic variable impedance prosthetic knee (VI Knee) was tested with five individuals (N = 5) with unilateral amputation above the knee at fixed speeds both above and below their normal walking speed. Subject gait was measured as they walked along an instrumented walkway via optical motion capture and force plates in the floor. Each subject's gait while using the VI Knee was compared to that while using their standard of care knee (OttoBock C-Leg). Significant differences (p < 0.05) in walking between the standard of care and variable impedance devices were seen in step length and hip range of motion symmetries, hip extension moment, knee power and torso lean angle. While using the VI Knee, several subjects demonstrated statistically significant improvements in gait, particularly in increased hip range of motion symmetry between affected and intact sides, greater prosthesis knee power and in reducing upper body involvement in the walking task by decreasing forward and affected side lean and reducing the pelvis-torso twist coupling. These changes to torso posture during gait also resulted in increased terminal stance hip flexion moment across subjects. Detriments to gait were also observed in that some subjects exhibited decreased step length symmetry while using the VI Knee compared to the C-Leg. The knee tested represents the potential to improve gait biomechanics and reduce upper body involvement in persons with above knee amputation compared to current standard of care devices. While using the VI Knee, subjects demonstrated statistically significant improvements in several aspects of gait though some were worsened while using the device. It is possible that these negative effects may be mitigated through longer term training and experience with the VI Knee. Given the demonstrated benefits and the potential to reduce or eliminate detriments through training, using a powered device like the VI Knee, particularly over an extended period of time, may help to improve walking performance and comfort.


Evolving Educational Techniques in Surgical Training.
Training competent and professional surgeons efficiently and effectively requires innovation and modernization of educational methods. Today's medical learner is quite adept at using multiple platforms to gain information, providing surgical educators with numerous innovative avenues to promote learning. With the growth of technology, and the restriction of work hours in surgical education, there has been an increase in use of simulation, including virtual reality, robotics, telemedicine, and gaming. The use of simulation has shifted the learning of basic surgical skills to the laboratory, reserving limited time in the operating room for the acquisition of complex surgical skills".


Preoperative taxane-based chemotherapy in a standardized protocol for locally advanced breast cancer.
To assess the feasibility of a standardized multidisciplinary protocol for the management of locally advanced breast cancer (LABC). We also evaluated the accuracy of magnetic resonance imaging (MRI) and positron emission tomography (PET) in predicting the extent of residual disease. Patients with LABC were offered preoperative chemotherapy of docetaxel 75 mg/m(2) , doxorubicin 50 mg/m(2) , cyclophosphamide 500 mg/m(2) (TAC), every 21 days for six cycles, until progression or intolerable toxicity. MRI and PET were performed at baseline and six cycles. Patients underwent a mastectomy or complete local excision, followed by radiotherapy. Trastuzumab and endocrine treatment were recommended where appropriate. Between April 2005 and October 2006, 51 patients were included from three institutions, and 50 received TAC (90% commenced within 35 days of diagnosis), with 44 patients completing six cycles (88%). Pathological complete response was seen in 10 patients (19.6%); all had invasive ductal carcinoma. No patient with invasive lobular carcinoma achieved pathological complete response. MRI was the most accurate method of assessing the extent of residual cancer. In total, 45 (88%) patients underwent surgery within the protocol-specified time and 12 (23%) patients had breast conservation surgery. At a median follow-up of 41.3 months, there were three local recurrences. Ten patients (19.6%) developed distant metastases, resulting in an 80% actuarial disease-free survival. This regimen of TAC is effective and well-tolerated and is likely to result in improved outcomes since patients can receive optimal multimodality treatments.


Vps9 family protein Muk1 is the second Rab5 guanosine nucleotide exchange factor in budding yeast.
VPS9 domains can act as guanosine nucleotide exchange factors (GEFs) against small G proteins of the Rab5 family. Saccharomyces cerevisiae vps9Δ mutants have trafficking defects considerably less severe than multiple deletions of the three cognate Rab5 paralogs (Vps21, Ypt52, and Ypt53). Here, we show that Muk1, which also contains a VPS9 domain, acts as a second GEF against Vps21, Ypt52, and Ypt53. Muk1 is partially redundant with Vps9 in vivo, with vps9Δ muk1Δ double mutant cells displaying hypersensitivity to temperature and ionic stress, as well as profound impairments in endocytic and Golgi endosome trafficking, including defects in sorting through the multivesicular body. Cells lacking both Vps9 and Muk1 closely phenocopy double and triple knock-out strains lacking Rab5 paralogs. Microscopy and overexpression experiments demonstrate that Vps9 and Muk1 have distinct localization determinants. These experiments establish Muk1 as the second Rab5 GEF in budding yeast.


Effects of RHAMM/HMMR-Selective Peptides on Survival of Breast Cancer Cells.
RHAMM-selective peptides in a concentration of 10 μg/ml (2×10(-7) M) inhibited the growth of MDA-MB-231 breast cancer cells over 48 h. Treatment of cancer cells with RHAMM-selective peptides induced apoptosis and necrosis and increased caspase-3 activity (by 30%).


Histidine-rich glycoprotein modulation of the anticoagulant activity of heparin. Evidence for a mechanism involving competition with both antithrombin and thrombin for heparin binding.
Heparin binding to rabbit histidine-rich glycoprotein (HRG) was studied in a purified system, allowing for determination of a heparin dissociation constant of approximately 5.5 X 10(-8) M for the interaction with HRG at pH 7.0. The strong interaction between heparin and HRG was demonstrated to be competitive with the binding of both antithrombin and thrombin to the heparin chain. HRG was further tested as a modulator of the anticoagulant activity of heparin by comparing rates of the heparin-catalyzed reaction between antithrombin and thrombin in the presence and absence of added HRG. The heparin-antithrombin-thrombin reaction was modeled using the formalism of a two-substrate enzyme-catalyzed reaction with heparin as the enzyme and HRG analyzed as an enzyme inhibitor. HRG was shown to compete with both antithrombin and thrombin for binding to heparin by this kinetic analysis. Thus, both the kinetic and heparin-binding data indicate that the mechanism by which HRG modulates heparin anticoagulant activity involves competition for heparin with both the inhibitor and the protease. Inhibition by HRG of the heparin-catalyzed reaction was found to be highly dependent on pH, with a sharp increase in inhibition from about 15% to greater than 90% observed as pH was lowered from 7.4 to 7.0. Since little change in the rate of the heparin-catalyzed inhibition of thrombin by antithrombin occurs in this pH region, the dramatic change in HRG inhibition seen upon pH titration may reflect increasing ionic interaction between heparin and HRG due to the protonation of histidine residues which occurs in this pH region.


Solubility and surface thermodynamics of conducting polymers by inverse gas chromatography. III: polypyrrole chloride.
Inverse gas chromatography, IGC, was applied to characterize conducting polypyrrole chloride (PPyCl) using twenty three solvents. IGC is able to reveal the change in the morphology, the strength of solvent-PPyCl interactions, thermodynamics parameters (χ12, Ω1(∞)), solvent and polymer solubility parameters, and molar heats of sorption, mixing and evaporation (ΔH1(s), ΔH1(∞), ΔH1(v)). The following solvents showed stronger interactions than others; yet, none of these solvents are good solvents for PPyCl: dodecane among the alkane family, tetrahydrofuran and methyl ethyl ketone among the oxy and keto group, dichloromethane among the chlorinated group up to 120°C and chloroform at 180°C, and toluene among the cyclic and aromatic group. Overall, the groups showed higher affinities to PPyCl are: acetates, oxy and cyclic, and chlorinated groups. Comprehensive solvents and PPyCl solubility parameters are obtained. The latter showed that PPyCl is not soluble in any solvent used.


Bovine pituitary intraglandular colloid fraction F5 localized in the rat endocrine pancreas.
Bovine pituitary intraglandular colloid thought to be a waste product, is the holocrine secretion of intermediate lobe cells. It is housed in the intraglandular lumen (residual lumen) and is extruded into the venous circulation of the cavernous sinuses via clefts in the capsule of the gland aligned with the intraglandular lumen. Intraglandular colloid, fraction F5 (mol.wt 34,000), radiolabeled with (125I)Na and injected (0.15 ml) into the right internal jugular vein of male Wistar rats, accumulated in the endocrine pancreas. Autoradiographs showed that the material had specifically localized in the capillary network of the endocrine pancreas. Since the intermediate lobe is poorly vascularized, intraglandular colloid is considered to be the transport medium for intermediate lobe materials.


Spontaneous granulation of moderately halophilic sludge inoculated with saltern sediments from single granule into multi-granule aggregation.
There is very limited research on the application of moderate halophiles for biotreatment of hypersaline wastewater widely generated from some industries. This study demonstrated the development of moderate halophiles inoculated from saltern sediments into aerobic granule sludge (AGS) to treat hypersaline wastewater with a salinity of 100 g/L. The granulation of moderate halophiles can occur without applying the settling velocity selective pressure. The saltern sediment initially aggregated into single small granules and finally developed into 1200 ± 50 μm multiparticle granules. The halophiles affiliated in Halomonas was dominant in the granular bacterial community, with a relative abundance of 94.52%. Halomonas ventosae secreted sulfated polysaccharides. The sulfated polysaccharides content accounted for 63.95 ± 2.10% in the polysaccharides (PS), having an adhesive role in connecting single granules. Multiparticle granules showed the clear stratified structure, with α-D-glucopyranose polysaccharides in the inner bounders and β-D-glucopyranose polysaccharides in the outer. The moderately granular sludge showed the stable chemical oxygen demand (COD) removal efficiency of >90% and the aerobic total inorganic nitrogen (TIN) removal efficiency (equal to ammonia removal) of 70 ± 5.00%. This paper contributes new insight into the formation of moderately halophilic granular sludge and accelerates the application of moderately halophilic granular sludge to treat hypersaline wastewater.


p63 expression in odontogenic cysts.
A total of 123 cases of odontogenic cysts, distributed as follows: 30 follicular (FC), 35 radicular (RC), 53 keratocysts, 1 glandular odontogenic and 4 calcifying odontogenic cysts, were analysed by immunohistochemistry for expression of p63, a component of p53 protein family. In RCs p63 positivity was not only in basal and parabasal layers but also in the intermediate layer and about 1/3 of cases displayed a percentage of stained cells comprised between 0 and <5%, and about 2/3 between >5% and <50%. In FCs positivity was confined to basal and parabasal layers of the epithelium and in the majority of FCs the stained cells were comprised between 0 and <5%. OKCs displayed the most intense and diffuse p63 labeling. In conclusion, these data suggest that p63 expression may be useful to identify cysts type with more aggressive and invasive phenotype supporting the hypothesis of a suprabasal proliferative compartment in OKCs.


The Asian-American variant of human papillomavirus type 16 exhibits higher activation of MAPK and PI3K/AKT signaling pathways, transformation, migration and invasion of primary human keratinocytes.
Asian-American (AA) HPV-16 variants are associated with higher risk of cancer. Abnormal activation of intracellular signaling play a critical role in cancer development and progression. Our aim was to elucidate mechanisms underlying the higher oncogenic potential attributed to AA variant. We evaluated activation of MAPK and PI3K/AKT pathways in primary human keratinocytes (PHKs) transduced with E6/E7 of three HPV-16 variants: E-P, AA, E-350G. Phenotypes examined included migration, anchorage independent growth and invasion. AA PHKs presented the highest levels of active proteins involved in all cascades analyzed: MAPK-ERK, MAPK-p38 and PI3K-AKT. AA PHKs were more efficient in promoting anchorage independent growth, and in stimulating cell migration and invasion. MEK1 inhibition decreased migration. The mesenchymal phenotype marker vimentin was increased in AA PHKs. Our results suggest that MEK1, ERK2, AKT2 hyperactivation influence cellular behavior by means of GSK-3b inactivation and EMT induction prompting AA immortalized PHKs to more efficiently surpass carcinogenesis steps.


Stigma toward Wuhan people during the COVID-19 epidemic: an exploratory study based on social media.
Stigma associated with infectious diseases is common and causes various negative effects on stigmatized people. With Wuhan as the center of the COVID-19 outbreak in China, its people were likely to be the target of stigmatization. To evaluate the severity of stigmatization toward Wuhan people and provide necessary information for stigma mitigation, this study aimed to identify the stigmatizing attitudes toward Wuhan people and trace their changes as COVID-19 progresses in China by analyzing related posts on social media. We collected 19,780 Weibo posts containing the keyword 'Wuhan people' and performed a content analysis to identify stigmatizing attitudes in the posts. Then, we divided our observation time into three periods and performed repeated-measures ANOVA to compare the differences in attitudes during the three periods. The results showed that stigma was mild, with 2.46% of related posts being stigmatizing. The percentages of stigmatizing posts differed significantly during the three periods. The percentages of 'Infectious' posts and 'Stupid' posts were significantly different for the three periods. The percentage of 'Irresponsible' posts was not significantly different for the three periods. After government interventions, stigma did not decrease significantly, and stigma with the 'Infectious' attitude even increased. It was not until the government interventions took effect that stigma significantly reduced. This study found that stigma toward Wuhan people included diverse attitudes and changed at different periods. After government interventions but before they took effect, stigma with the 'Infectious' attitude increased. After government interventions took effect, general stigma and stigmas with 'Infectious' and 'Stupid' attitudes decreased. This study constituted an important endeavor to understand the stigma toward Wuhan people in China during the COVID-19 epidemic. Implications for stigma reduction and improvement of the public's perception during different periods of epidemic control are discussed.


Cell signalling pathway involved in PACAP-induced AR4-2J cell proliferation.
The novel 38-amino acid neuropeptide PACAP (pituitary adenylate activating peptide) has recently been shown to induce the pancreatic acinar tumour AR4-2J cell growth. This growth promoting effect of PACAP was, however, independent of adenylate cyclase activation but suppressed by pertussis toxin and the somatostatin analog SMS 201-995. This study was undertaken to search for potential cell signalling pathways involved in the growth promoting effect of PACAP on AR4-2J cells. The AR4-2J cells were grown in Dulbecco's Modified Eagle's Medium containing 10% foetal calf serum. For studies on cell signalling pathways, all experiments were carried out on cells which have reached 50 to 75% confluency. At that point, they were transferred to serum free medium overnight with or without 1 microCi/ml myristic acid. The next morning, cells were harvested, washed and used for tyrosine kinase and phospholipase D (PLD) activities. For studies on growth, cells were grown for 2 days in the presence of 1 nM PACAP +/- the different inhibitors of tyrosine kinase and PLD. PACAP-38 and -27 caused a dose-dependent and parallel activation of tyrosine kinase and PLD an effect prevented by the antagonist PACAP 7-38. PACAP-38-stimulated tyrosine kinase and PLD activation are both dose-dependently inhibited by SMS 201-995. Finally, PACAP-stimulated tyrosine kinase and PLD activities are both inhibited by cell's preincubation with genistein and pertussis toxin. After 2 days, the PACAP-induced increase in AR4-2J cell growth was significantly inhibited by increasing concentrations of genistein and wortmannin, inhibitors of tyrosine kinase, PLD and phosphatidylinositol 3-kinase, respectively. PACAP can induce concomitant activation of tyrosine kinase and PLD; this finding and the observation that inhibition of these two enzymes inhibited PACAP-induced AR4-2J cell growth strongly suggests that they are intimately involved in the overall process of PACAP-induced AR4-2J cell proliferation.


Recent applications of quantitative electroencephalography in adult intensive care units: a comprehensive review.
Quantitative electroencephalography (qEEG) refers to the numerical analysis and/or visual transformations of raw electroencephalography (EEG) signals. Evaluation of qEEG in intensive care units (ICU) faces unique challenges that warrant investigation separate from those conducted in other settings. Additionally, the pathophysiology, management, and EEG patterns of critically ill conditions often significantly differ between adults and children. Thus, it is important to distinguish the literature on qEEGs specifically performed in adult ICUs. The aim of this review is to summarize the studies using qEEG for clinical evaluation of patients in adult ICUs performed over the past decade (since 2010), and to present the state of the art of these techniques. Overall, these studies have reported that qEEG can reveal important information faster than typically possible with traditional methods of reviewing the raw EEG only, with reasonable accuracy. However, it is crucial to emphasize that qEEG must be reviewed in conjunction with raw EEG and in context of understanding the patients' clinical status. Because each qEEG panel only focuses on a few aspects of the entire EEG, different combinations of qEEG panels may be required for optimal analyses of each medical condition and individual patient. Currently in practical terms, qEEG can serve as a complementary, valuable tool for portions of the EEG that require more detailed review. Further multi-center collaborative studies are needed to ultimately develop standardized methods of employing qEEG that are generalizable across institutions. As qEEG techniques continue to advance, including those involving machine learning, qEEG will further benefit from algorithms specifically suited for ICUs.


Mechanical properties and collagen fiber orientation of tendon in young and elderly.
The purpose of this study was to investigate differences in the mechanical properties and collagen fiber orientation of tendon structures between young and elderly groups. The mechanical properties of tendon structures in medial gastrocnemius muscle were measured using ultrasonography during ramp and ballistic contractions. Tendon collagen fiber orientation was estimated from coefficient of variation (CV) of echogenicity on transverse ultrasonic images of Achilles tendon. Differences in elongation between ramp and ballistic contractions of elderly were significantly smaller than those of young group at 20-80% of MVC. During ramp contraction, hysteresis of elderly was significantly higher than that of young, whereas no difference in hysteresis during ballistic contraction was found between the two groups. Difference in hysteresis between ramp and ballistic contractions of elderly tended to be lower than that of young group. Mean echogenicity of elderly was significantly higher than that of young group, whereas no difference in CV of echogenicity was found between the two groups. These results suggest that smaller differences in elongation and hysteresis between ramp and ballistic contractions of elderly may be related to decreased water content within tendons. Furthermore, no difference in collagen fiber orientation of tendons was noted between the two groups.


Oral vinorelbine: feasibility and safety profile.
Patient preference as well as concerns and difficulties with intravenous access and pharmaco-economic issues have driven the development of oral vinorelbine. Four phase II studies were conducted in chemotherapy-naive non-small-cell lung cancer (NSCLC) and as first-line chemotherapy of advanced breast cancer (ABC). As recommended in the phase I dose-finding study, the first step used a weekly dose of 80 mg/m2. This regimen was associated with an excessive rate of early deaths (10%) due to complicated neutropenia and led to discontinuation of the first two studies. In a second step, the dose of 60 mg/m2/week was given for the first three courses and subsequently increased to 80 mg/m2/week, in the absence of severe neutropenia. One hundred and thirty eight patients (76 with NSCLC and 62 with ABC) received this regimen, of whom only five were unable to undergo dose escalation. The incidence of febrile neutropenia and neutropenic sepsis were low (2.9 and 3.6%, respectively). Although severe events were uncommon, nausea/vomiting and diarrhoea were frequent and primary prophylaxis with antiemetics should be recommended. Overall, the safety profile of oral vinorelbine at 60 mg/m2/week for the first three courses with escalation to 80 mg/m2 is qualitatively comparable to that of i.v. vinorelbine at standard doses. Similarly to i.v. chemotherapy, close haematological monitoring is necessary.


Imaging and pharmacokinetics of (64)Cu-DOTA-HB22.7 administered by intravenous, intraperitoneal, or subcutaneous injection to mice bearing non-Hodgkin's lymphoma xenografts.
The aim of the study is to compare the tumor-specific targeting, pharmacokinetics, and biodistribution of (64)Cu-DOTA-HB22.7 when administered to xenograft-bearing mice intravenously (IV), intraperitoneally (IP), and subcutaneously (SQ). Mice bearing human non-Hodgkin's lymphoma (NHL) xenografts were injected IV, IP, or SQ with (64)Cu-DOTA-HB22.7. Xenograft targeting was evaluated by micro positron emission tomography (microPET) and confirmed by organ biodistribution studies. Blood measurements of (64)Cu were performed to determine the pharmacokinetics and clearance of (64)Cu-DOTA-HB22.7. (64)Cu-DOTA-HB22.7 demonstrated equivalent tumor targeting within 24-48 h, regardless of the route of administration. Organ biodistribution confirmed tumor-specific targeting. Blood pharmacokinetics demonstrated that (64)Cu-DOTA-HB22.7 accessed the bloodstream after IP and SQ administration to a similar degree as IV administration, albeit at a slower rate. These findings establish (64)Cu-DOTA-HB22.7 as a potential radioimmunotherapeutic and/or NHL-specific imaging agent. These findings provide evidence that IP and SQ administration can achieve results equivalent to IV administration and may lead to more efficient, reproducible treatment plans for antibody-based therapeutics.


Efficacy of the subcutaneous reformulated triptorelin depot in children with central precocious puberty.
The efficacy, safety and acceptance of newly formulated triptorelin s.c. (Decapeptyl Depot, DDsc) was compared to triptorelin i.m. (DDim) in seven children with central precocious puberty (CPP) in a prospective study. Both formulations were given for 6 months consecutively. During both treatment periods suppression of basal and gonadotropin-releasing hormone (GnRH)-stimulated levels of luteinizing hormone and follicle-stimulating hormone, suppression of sex steroids, arrest of the maturation of gonads and uterus, and slowing of bone maturation were achieved. The height standard deviation score for bone age increased significantly during DDsc treatment (-1.33+/-0.90 to 1.07+/-0.92, p < 0.05). The ratio bone age/chronological age decreased significantly during both treatment periods (1.25+/-0.24 to 1.20+/-0.23,p < 0.05, and 1.20+/-0.23 to 1.16+/-0.22, p < 0.05). With the injection of DDsc in the abdominal wall, a palpable, non-irritating resistance which gradually decreased in size occurred in one patient. With injection into the thigh no indurations were seen. No allergic reactions were encountered. Five of the patients considered DDsc therapy as more pleasant and described a definite decrease in fear of injections. All parents considered DDsc treatment as equal or better than DDim in respect to the suppression of their child' s puberty. It was concluded that DDsc is equipotent to DDim in the treatment of CPP. For the majority of patients DDsc was more acceptable than DDim. The thigh is recommended for the subcutaneous application of DD.


Effect of donor pancreas extraction time on pancreas transplantation-a Swiss tertiary center experience.
We aimed to assess the effect of donor pancreas extraction time (ET) on postoperative complications and graft function after pancreas transplantation (PT). We analyzed all consecutive donor pancreas procurements for the simultaneous pancreas and kidney transplantation (SPK) and the associated PT in a Swiss transplant center over a 20-year period. Pancreas ET was defined as the time from cold flush to static storage of the pancreas on ice. The primary endpoint was the effect of extraction time on surgical complications. Secondary endpoints comprised the effect of ET on graft function (insulin-free survival) and graft pancreatitis. Of 115 procured pancreas grafts the median donor pancreas ET was 65 min (IQR: 48-78 min). In multivariable analysis, ET did not negatively affect major complications (OR 1.41 [95% CI: .59-3.36]; p = .438) and insulin-free survival (HR 1.42 [95% CI: .55-3.63]; p = .459). The median CIT was 522 (441-608) min. CIT was associated with major complications (OR 2.51 [95% CI: 1.11-5.68]; p = .027), but without impact on insulin-free survival (HR 1.94 [95% CI: .84-4.48]; p = .119). Patients with and without graft pancreatitis had no statistically significant differences in ET and CIT (p = .164 and p = .47, respectively). In multivariable analysis, Amylase levels > 270 U/L on postoperative day 1 were significantly associated with major complications (OR 3.61 [95% CI: 1.06-12.32]; p = .040). Our results suggest that although no effect of ET on complications and graft function after PT was found, shorter CIT and less graft pancreatitis can have a positive impact on surgical complications. Results could possibly be influenced by the exceptional quality of the pancreas donors, with short travel distances and preservation times in Switzerland.


Presenilin-1 interacts directly with the beta-site amyloid protein precursor cleaving enzyme (BACE1).
A neuropathological hallmark of Alzheimer's disease is the presence of amyloid plaques. The major constituent of these plaques, occurring largely in brain areas important for memory and cognition, is the 40-42 amyloid residues (Abeta). Abeta is derived from the amyloid protein precursor after cleavage by the recently identified beta-secretase (BACE1) and the putative gamma-secretase complex containing presenilin 1 (PS1). In an attempt to develop a functional secretase enzymatic assay in yeast we demonstrate a direct binding between BACE1 and PS1. This interaction was confirmed in vivo using coimmunoprecipitation and colocalization studies in human cultured cells. Our results show that PS1 preferably binds immature BACE1, thus possibly acting as a functional regulator of BACE1 maturation and/or activity.


The use of ethylene oxide and low temperature steam/formaldehyde in hospitals.
Reprocessing of heat-labile medical equipment is a major problem in hospital practice. Requirements are either for sterilization, e.g. cardiac catheters, or disinfection, e.g. respiratory equipment. Methods of low temperature sterilization include ethylene oxide, low-temperature steam and formaldehyde (LTS/F) and immersion in 2% glutaraldehyde. Ethylene oxide is reliable, but potentially toxic. LTS/F machines operate at a high temperature, but treated equipment needs less aeration. The advantages of these processes and possible alternatives are discussed.


Insulin-like growth factor-2 antisense oligonucleotides inhibits myopia by expression blocking of retinal insulin-like growth factor-2 in guinea pig.
To clarify the role of IGF-2 on the development of myopia, the dynamic expression of IGF-2 was investigated in the FD eyes' retina, and the effects of intravitreous injection with IGF-2 ASON was studied on the diopter and axial eye length of FD eyes. 64 guinea pigs were divided into 2 groups. In group A (n = 24), the right eyes were covered. On days 7, 14 and 21, the diopter, axial eye length and level of IGF-2 of both eyes were measured in every 8 guinea pigs. In group B (n = 40), the right eyes were covered. On day 1, the right eyes were received intravitreal injection with 40 µg IGF-2SON, 10 µg, 20 µg or 40 µg IGF-2 ASON. The diopter, axial eye length and level of IGF-2 were measured on day 14. FD eyes showed myopic shift, axial length enlongation, and up-regulation in retinal IGF-2 from day 7 to day 21. The level of retinal IGF-2 in FD eyes was higher than that in non-FD eyes. Compare with FD eyes without injection, the myopia diopter of FD eyes decreased in received intravitreous injection with IGF-2 ASON, axial length shortened, and down-regulated with retinal IGF-2. With the increase dose of IGF-2 ASON, the change of myopic diopter, axial length, and level of retinal IGF-2 were showed more and more significant. FD is effective to up-regulate the level of retinal IGF-2 expression in guinea pig. Intravitreous injection with IGF-2 ASON can inhibit the development of myopia.


A facile and green pretreatment method for nonionic total organic halogen (NTOX) analysis in water - Step II. Using photolysis to convert NTOX completely into halides.
Adsorbable organic halogen (AOX) is a parameter conventionally used to indicate the sum of organic halogenated disinfection byproducts (DBPs), which are formed from the reactions of disinfectants with dissolved organic matter, bromide and iodide in water. To overcome the issues of the AOX analytical method, we proposed a new facile and green pretreatment method to enable the analysis of nonionic total organic halogen (NTOX) via the following three steps: 1) separation of NTOX and halides with electrodialysis, 2) conversion of NTOX with ultraviolet (UV) photolysis, and 3) analysis of halides with ion chromatography. To verify this proposal, we mainly evaluated the efficiency of vacuum ultraviolet (VUV) coupled with UV photolysis (VUV-UV) in converting NTOX into halides. Results showed that by applying VUV irradiation for 60 min and UV irradiation at pH 10-11 for another 30 min, over 85.5% of each halide from 20 representative small molecular weight DBPs (each at 100 μg-X/L level) was recovered. The purpose of UV photolysis under alkaline conditions was to reduce oxyhalides (such as bromate and iodate) formed in the VUV process back to halides. With the aid of electrospray ionization-triple quadrupole mass spectrometry, we captured the whole pictures of high molecular weight polar DBPs in a chlorinated drinking water before and after VUV-UV, through which averagely 96.4% of dehalogenation with the VUV-UV treatment was observed. An illustrative comparison of the conventional AOX method and the proposed NTOX method indicates that although the detected NTOX was lower (by 2.3-30.6%) than AOX, the results of the two methods were highly correlated (R2 > 0.97). All these hence verified the photolysis as a mature yet novel tool for sample pretreatment in environmental analytical chemistry.


Self-perceived and independently observed competence of young adolescents as a function of parental marital conflict and recent divorce.
The self-perceived and independently observed cognitive and social competence of young adolescents as a function of parental conflict and recent divorce was investigated. Subjects were 40 young adolescents between the ages of 11 years 1 month and 15 years 1 month. A 2 X 2 factorial design was used, with the independent variables being parental marital status (married vs. recently divorced) and parental conflict (high vs. low). Dependent variables included the following measures of adolescent competence: adolescent-completed measures of self-perceived competence, teacher-completed measures, behavioral observations, and school grades. The results indicated that the level of parental conflict, rather than parental marital status, appears to be the critical variable associated with adolescents' independently observed levels of cognitive and social competence. In regard to adolescents' self-perceived levels of cognitive and social competence, parental marital status was found to be the critical variable. The implications of these findings are discussed.


Autistic spectrum disorder (ASD) - Gene, molecular and pathway signatures linking systemic inflammation, mitochondrial dysfunction, transsynaptic signalling, and neurodevelopment.
Despite advances in autism spectrum disorder (ASD) research and the vast genomic, transcriptomic, and proteomic data available, there are still controversies regarding the pathways and molecular signatures underlying the neurodevelopmental disorders leading to ASD. To delineate these underpinning signatures, we examined the two largest gene expression meta-analysis datasets obtained from the brain and peripheral blood mononuclear cells (PBMCs) of 1355 ASD patients and 1110 controls. We performed network, enrichment, and annotation analyses using the differentially expressed genes, transcripts, and proteins identified in ASD patients. Transcription factor network analyses in up- and down-regulated genes in brain tissue and PBMCs in ASD showed eight main transcription factors, namely: BCL3, CEBPB, IRF1, IRF8, KAT2A, NELFE, RELA, and TRIM28. The upregulated gene networks in PBMCs of ASD patients are strongly associated with activated immune-inflammatory pathways, including interferon-α signaling, and cellular responses to DNA repair. Enrichment analyses of the upregulated CNS gene networks indicate involvement of immune-inflammatory pathways, cytokine production, Toll-Like Receptor signalling, with a major involvement of the PI3K-Akt pathway. Analyses of the downregulated CNS genes suggest electron transport chain dysfunctions at multiple levels. Network topological analyses revealed that the consequent aberrations in axonogenesis, neurogenesis, synaptic transmission, and regulation of transsynaptic signalling affect neurodevelopment with subsequent impairments in social behaviours and neurocognition. The results suggest a defense response against viral infection. Peripheral activation of immune-inflammatory pathways, most likely induced by viral infections, may result in CNS neuroinflammation and mitochondrial dysfunction, leading to abnormalities in transsynaptic transmission, and brain neurodevelopment.


Maternal and foetal immune responses of cattle following an experimental challenge with Neospora caninum at day 70 of gestation.
The immune responses of pregnant cattle and their foetuses were examined following inoculation on day 70 of gestation either intravenously (iv) (group 1) or subcutaneously (sc) (group 2) with live NC1 strain tachyzoites or with Vero cells (control) (group 3). Peripheral blood mononuclear cell (PBMC) responses to Neospora antigen and foetal viability were assessed throughout the experiment. Two animals from each group were sacrificed at 14, 28, 42 and 56 days post inoculation (pi). At post mortem, maternal lymph nodes, spleen and PBMC and when possible foetal spleen, thymus and PBMC samples were collected for analysis. Inoculation with NC1 (iv and sc) lead to foetal deaths in all group 1 dams (6/6) and in 3/6 group 2 dams from day 28pi; statistically significant (p ≤ 0.05) increases in cell-mediated immune (CMI) responses including antigen-specific cell proliferation and IFN-γ production as well as increased levels of IL-4, IL-10 and IL-12 were observed in challenged dams compared to the group 3 animals. Lymph node samples from the group 2 animals carrying live foetuses showed greater levels of cellular proliferation as well as significantly (p ≤ 0.05) higher levels of IFN-γ compared to the dams in group 2 carrying dead foetuses. Foetal spleen, thymus and PBMC samples demonstrated cellular proliferation as well as IFN-γ, IL-4, IL-10 and IL-12 production following mitogenic stimulation with Con A from day 14pi (day 84 gestation) onwards. This study shows that the generation of robust peripheral and local maternal CMI responses (lymphoproliferation, IFN-γ) may inhibit the vertical transmission of the parasite.


Self-reported chemical sensitivity in Germany: a population-based survey.
Environmental clinics are frequented by patients with fears and complaints related to environmental triggers. A dose-independent overreaction to small doses of widely used and generally non-toxic chemicals is referred to as multiple chemical sensitivity (MCS), but no clearly defined clinical syndrome with objective physical findings has been delineated so far. We aimed to obtain information about symptoms, supposed environmental triggers, the frequency of self-reported chemical sensitivity, and of the diagnosis MCS in Germany. We conducted a representative survey among 2032 adult Germans. We found self-reported chemical sensitivity in 9% and physician-diagnosed MCS in 0.5% of our representative sample. Physical complaints were common in the whole study population and in chemically sensitive individuals, but there was no clear-cut symptom constellation among the latter. The most common complaints were headache, fatigue, sleep disturbances, joint pain, mood changes and nervousness. A subjective connection between complaints and environmental triggers was denied by 67% of the whole group and by 35% of the self-reported chemically sensitive. Factor analysis of environmental triggers suggested that a specific exposure situation rather than chemical similarity is the basis for individual trigger combinations. The prevalence of subjective sensitivity towards chemicals is similar to such rates reported from other countries. There is a relatively low awareness of the MCS-concept, and it appears to be diagnosed less frequently than, e.g., in the USA. Since symptoms and triggers in chemically sensitive individuals did not differ from the general population, our data do not suggest the existence of a widespread new syndrome related to chemical sensitivities in Germany. We outline the limitations of self-reported chemical sensitivity as the major criterion for such a contentious diagnosis as MCS.


Risk factors of early death in pediatric hemophagocytic lymphohistocytosis: Retrospective cohort study.
Hemophagocytic lymphocytosis (HLH) is a rare life-threatening hyperinflammatory syndrome in which early mortality remains high in patients with HLH. We retrospectively collected the medical records of all pediatric patients diagnosed with HLH at the West China Second Hospital of Sichuan University between January 2014 and December 2020. Collect demographic, laboratory, clinical, genetic profiles, treatment information and perform statistical analysis from records. Risk factors for death 30 days after admission were evaluated using a multivariable logistic regression model. A total of 110 pediatric HLH patients were enrolled. The median age of patients was 44 months (IQR 23-100.5) and 62 (56.4%) in males. The 30-day mortality rate for admission to this cohort was 34 (30.9%). Multivariate logistic regression analysis showed that heart failure (OR = 13.389, 95% CI, 1.671-107.256, p = 0.015) and hypoproteinemia (OR = 4.841, 95% CI, 1.282-18.288, p = 0.020) were associated with increased early mortality in children with HLH. These identified risk factors may help clinicians stratify patients with HLH and develop targeted treatment strategies. More research is needed to explore the best treatment strategies for patients with HLH to reduce early mortality in patients with HLH.


Orally bioavailable highly potent HIV protease inhibitors against PI-resistant virus.
Efforts directed to identifying potent HIV protease inhibitors (PI) have yielded a class of compounds that are not only very active against wild-type (NL4-3) HIV virus but also very potent against a panel of PI-resistant viral isolates. Chemistry and biology are described.


Cost-utility analysis of interferon-free treatments for patients with early-stage genotype 1 hepatitis C virus in Brazil.
We conducted a cost-utility analysis of available interferon-free treatments for patients with early-stage genotype 1 chronic hepatitis C based on a Brazilian public health system perspective. A Markov model was derived using a cohort of stage F0-F2 patients treated as recommended by the Brazilian national guidelines. Glecaprevir plus pibrentasvir was superior to all other treatments, followed by sofosbuvir plus velpatasvir. Sofosbuvir plus daclatasvir was identified as the least cost-effective option. The above findings were confirmed via probabilistic sensitivity analysis and the tested scenarios.


Cytocompatible asymmetrical coating for Janus carrier synthesis through capillary wetting and ascending.
Despite the successful implementation of elegant strategies for the fabrication of Janus microstructures, two critical factors have limited the applicability of most techniques for the partial modification of living cell surfaces: harsh conditions that could disintegrate cells, and the lack of an effective route to accomplish a mild modification for living cells. In this study, an expeditious synthesis, named lower-half occupation by capillary ascended liquids (LOCAL), is proposed for the fabrication of asymmetrical structures surrounding not only microbeads but also both living adherent and buoyant mammalian cells. The microbeads or living cells are safely supported and trapped on the apical sides of a micropillar array, which prevents them from contacting the bottom substrate. As the coating agents further transfer and contact the trapped particles through interpillar capillary flow, the autonomous capillary ascending coats the free bottom surfaces of the target particles within 2 min, with significantly small quantities of coating agents. The self-assembled architectures of the cells demonstrate thoroughly maintained cell viability, highlighting the potential of the LOCAL method as a desirable alternative to the widely applied state-of-art methods for developing Janus beads and Janus cells.


Best Practices in Surveillance for Barrett's Esophagus.
Barrett's esophagus is the precursor lesion for esophageal adenocarcinoma. The goals of endoscopic surveillance are to detect dysplasia and early esophageal adenocarcinoma in order to improve patient outcomes. Despite the ongoing debate regarding the efficacy of surveillance, all current gastrointestinal societies recommend surveillance at this time. Optimal surveillance technique includes adequate inspection time, evaluation using high-definition white light and chromoendoscopy, appropriate documentation of the metaplastic segment using the Prague C & M criteria as well as the Paris classification should lesions be found, utilization of the Seattle biopsy protocol, and endoscopic resection of visible lesions.


The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results.
The embryonic cardiomyocyte cell line H9C2 is commonly used in numerous in vitro studies, including cardiotoxicity analyses of new drugs. So far no results were published for studies on cell parameters variability during the cell line ageing process. For this reason the aim of the study was to evaluate the effect of a number of H9C2 rat embryonic cardiomyocytes passages on repeatability of study results for selected cytotoxicity parameters, with doxorubicin as a model toxic agent. The cultures were passaged twenty-five times. Cells from passage 1, 5, 10, 15, 20 and 25 were treated with doxorubicin for 24 h. Then drug cytotoxicity was evaluated with the MTT test and additionally the nuclear factor erythroid 2-related factor (Nrf2) gene expression was examined. The analysis of oxidative stress intensity and cell morphology was also assessed. The microscopic appearance of cells indicates that untreated cardiomyocytes morphology changes as well as sensitivity to toxic effects increases with the number of passages. Also an increase in oxidative stress in cells occurs with further passaging of cardiomyocytes. Statistical significance of differences in conducted tests results depended on doxorubicin concentration but in many cases the H9C2 line was found to be a reliable in vitro model only for the first five passages. For this reason it is important to take into consideration that further culturing of cardiomyocytes may not ensure repeatability of study results due to the culture ageing.


Heparanase DNA vaccine delivered by electroporation induces humoral immunity and cytoimmunity in animal models.
Electroporation (EP)-assisted DNA vaccination has been proven effective as an approach to the treatment of cancer. Although heparanase (HPA) is a potential target for patients with advanced tumor diseases, the efficacy of immunotherapeutic strategies targeting HPA has never been evaluated. In this study, humoral immunity was elicited using genetic vaccinations between C57BL/6J mice and Macaca fascicularis. The immunized serum neutralized HPA activity and attenuated the invasion of B16 cells in vitro. In addition, T lymphocytes from the splenic cells of the immunized mice induced HPA-specific cytotoxic lymphocytes (CTLs), which verified cytoimmunity. Prophylactic vaccination significantly suppressed tumor growth and metastasis in vivo and prolonged the survival rate in tumor-bearing murine models. In addition, RT-PCR and Western blot analyses of the primary tumors indicated less proliferation and angiogenesis and more apoptosis in the HPA-immunization immunotherapy groups. Simultaneously, the levels of IL-2, IL-4, and IFN-γ were not significantly greater in the HPA-immunized group than in PBS controls. Thus, we conclude that the combination of an anti-HPA antibody and a CTL response in HPA-immunization gene therapy is enough to attenuate tumor growth and metastasis. This is the first time that a DNA vaccine targeting HPA immunization assisted by EP has induced humoral immunity and cytoimmunity in vivo. This provides a basis for the continued development of DNA vaccines targeting HPA and the use of such vaccines in clinical settings.


[Prenatal diagnosis by amniocentesis. Clinical and cytogenetic experience in 1,500 cases].
Genetic amniocentesis is performed in México 25 years ago but only few works have been published. To analyze clinical and cytogenetic findings in consecutive patients submitted to genetic amniocentesis. An analysis was made of the clinical features, amniocentesis results and pregnancy outcome in 1500 consecutive cases of genetic amniocentesis. Sixty-eight fetuses with chromosomopathy (4.5%) were detected and two, with an inborn error of metabolism. The most frequent abnormalities were trisomy 21 (32 cases), trisomy 18 (10 cases), trisomy 13(6 cases), 45,X (6 cases), 47,XXY (4 cases). Pregnancy outcome is known in 474 patients (32%). There were five fetal losses (1%). Of the 68 cases with chromosomopathy, the outcome is known in 45, of which, 29 (64%) decided to have an abortion while 16 (35%) continued the pregnancy, six had a spontaneous abortion or perinatal death and ten had an alive new born. Among fetuses with normal or balanced karyotype and normal ultrasound, 11 out of 419 (2.6%) had congenital anomalies. Two of them had a condition known to be related with epigenetic regulation, (Russell Silver and Angelman syndrome). Amniocentesis is a reliable and low risk method. Cytogenetic findings in this series are similar to those reported in the literature. Most patients with fetal disease decided to have an abortion. The finding of two patients with a condition related with abnormal epigenetic regulation suggests that the magnitude of this risk remains to be defined.


Characterization of histamine H3 receptors in mouse brain using the H3 antagonist [125I]iodophenpropit.
We have characterized the binding of the histamine H3 receptor antagonist [125I]iodophenpropit to mouse brain. [125I]Iodophenpropit saturably bound to mouse brain membranes with a pKd-value of 9.31+/-0.04 nM and a receptor binding density of 290+/-8 fmol per mg protein. Saturation binding analysis revealed binding of [125I]iodophenpropit to a single class of sites, showing linear Scatchard plots and Hill coefficients not different from unity (nH=0.98+/-0.02). At a concentration of 0.25 nM [125I]iodophenpropit, specific binding represented about 75% of the total binding. Competition binding curves for H3 receptor antagonists were fitted best to a one-site model, showing pKi-values in general accordance with the pA2-values obtained in mouse cerebral cortex. Displacement of [125I]iodophenpropit by the H3 receptor agonists (R)-alpha-methylhistamine, immepip, imetit and histamine were fitted best to a two-site model. Competition binding curves of (R)-alpha-methylhistamine showed a rightward shift upon incubation with GTPgammaS (10 microM), indicating the involvement of G-proteins in H3 agonist binding. In contrast, competition binding curves of the antagonists iodophenpropit, thioperamide and burimamide were not affected by GTPgammaS (10 microM). Autoradiographic experiments showed that [125I]iodophenpropit binding sites were heterogeneously distributed, similarly to the distribution of histamine H3 receptors reported in rat brain. Highest densities were observed in the cerebral cortex, the striatum, the nucleus accumbens, the globus pallidus and the substantia nigra. In conclusion, we have demonstrated that in mouse brain, [125I]iodophenpropit selectively binds to histamine H3 receptors. We also observed that the mouse brain H3 receptors labelled by [125I]iodophenpropit displayed binding characteristics and a distribution similar to rat brain.


Application of fused-grid-based CYP-Template systems for genotoxic substances to understand the metabolisms.
Understanding of metabolic processes is a key factor to evaluate biological effects of carcinogen and mutagens. Applicability of fused-grid Template* systems of CYP enzymes (Drug Metab Pharmacokinet 2019, 2020, 2021, and 2022) was tested for three phenomena. (1) Possible causal relationships between CYP-mediated metabolisms of β-naphthoflavone and 3-methylcholanthrene and the high inducibility of CYP enzymes were examined. Selective involvement of non-constitutive CYP1A1, but not constitutive CYP1A2, was suggested on the oxidative metabolisms of efficient inducers, β-naphthoflavone and 3-methylcholanthrene. These results supported the view of the causal link of their high inducibility with their inefficient metabolisms due to the lack of CYP1A1 in livers at early periods after the administration of both inducers. (2) Clear differences exist between human and rodent CYP1A1 enzymes on their catalyses with heterocyclic amines, dioxins and polyaromatic hydrocarbons (PAHs). Reciprocal comparison of simulation results with experimental data suggested the rodent specific site and distinct sitting-preferences of ligands on Template for human and rodent CYP1A1 enzymes. (3) Enhancement of metabolic activation and co-mutagenicity have been known as phenomena associated with Salmonella mutagenesis assay. Both the phenomena were examined on CYP-Templates in ways of simultaneous bi-molecule bindings of distinct ligands as trigger and pro-metabolized molecules. α-Naphthoflavone and norharman served consistently as trigger-molecules to support the oxidations of PAHs and arylamines sitting simultaneously as pro-metabolized molecules on Templates of CYP1A1, CYP1A2 and CYP3A4. These CYP-Template simulation systems with deciphering capabilities are promising tools to understand the mechanism basis of metabolic activations and to support confident judgements in safety assessments.


Designing Dual-functionalized Gels for Self-reconfiguration and Autonomous Motion.
Human motion is enabled by the concerted expansion and contraction of interconnected muscles that are powered by inherent biochemical reactions. One of the challenges in the field of biomimicry is eliciting this form of motion from purely synthetic materials, which typically do not generate internalized reactions to drive mechanical action. Moreover, for practical applications, this bio-inspired motion must be readily controllable. Herein, we develop a computational model to design a new class of polymer gels where structural reconfigurations and internalized reactions are intimately linked to produce autonomous motion, which can be directed with light. These gels contain both spirobenzopyran (SP) chromophores and the ruthenium catalysts that drive the oscillatory Belousov-Zhabotinsky (BZ) reaction. Importantly, both the SP moieties and the BZ reaction are photosensitive. When these dual-functionalized gels are exposed to non-uniform illumination, the localized contraction of the gel (due to the SP moieties) in the presence of traveling chemical waves (due to the BZ reaction) leads to new forms of spontaneous, self-sustained movement, which cannot be achieved by either of the mono-functionalized networks.


A new dataset of pheromone and pheromone-gene structures from the ciliate, Euplotes crassus.
Like many other organisms, ciliates communicate and interact socially via diffusible chemical signals, named pheromones, that are functionally associated with a genetic mating-type mechanism of cell self/not-self recognition. In Euplotes species, pheromones form species-specific families of small, globular, and disulfide-rich proteins folding into exclusively helical secondary structures. Each is specified by one of a series of high-multiple alleles that are inherited in Mendelian fashion with relationships of co-dominance at the so-called mat genetic locus of the cell transcriptionally inert micronuclear genome, and expressed in the transcriptionally active macronuclear genome as individual DNA molecules in which the central coding region is flanked by 5'-leader and 3'-trailer noncoding regions ending with C4A4/T4G4 telomeric repeats. In E. crassus, a cosmopolitan marine species with a long tradition in the study of ciliate mating systems and breeding patterns, oligonucleotides specific to amino acid sequences of pheromones Ec-1 and Ec-α were previously used to clone and sequence a first set of four structurally distinct macronuclear (mac) pheromone coding genes, mac-ec-α, mac-ec-1, mac-ec-2 and mac-ec-3, from two interbreeding strains, L-2D and POR-73. The use of these oligonucleotides in PCR amplifications of macronuclear DNA preparations from three other E. crassus interbreeding strains, ES10, Fava4 and MN4, has now resulted in the characterization of a second set of eight new pheromone coding genes, mac-ec-β, mac-ec-γ, mac-ec-δ, mac-ec-ε, mac-ec-µ, mac-ec-4, mac-ec-5 and mac-ec-6. Multiple alignment between previously and newly determined pheromone-gene sequences reinforces the concept that the E. crassus pheromone-gene family includes two sub-families, which likely reflect a duplication of the micronuclear mat gene locus and represent an apomorphic trait of the E. crassus clade. Members of one sub-family (each identified with a Greek letter) show a 500-bp 5'-leader noncoding region rich in AGGA/AGGGA repetitions, and encode 56-amino acid pheromones with eight conserved Cys residues. Members of the other sub-family (each identified with an Arabic numeral) show an 800-bp 5'-leader noncoding region without AGGA/AGGGA repetitions, and encode 45-amino acid pheromones with ten conserved Cys residues.


Comparative evaluation of zinc oxide-eugenol and zinc oxide with Neem oil in root canal treatment of primary teeth: Split-mouth study with 12 months follow-up.
Various root canal filling materials are used to preserve pulp tissue involved carious the primary tooth. A single material that fulfills all the requirements of an ideal root canal filling material for primary teeth is yet unavailable. Hence, this study was initiated to evaluate clinically and radiographically, the efficacy of two obturating materials - zinc oxide eugenol (ZOE) and zinc oxide mixed with Neem oil (ZON) for 12 months. To assess the success rate of a mixture of zinc oxide and neem oil in comparison to zinc oxide-eugenol paste, as an obturating material in primary molars. To compare the efficacy of ZON as an obturating material at the various time interval of 3, 6, 9, and 12 months postoperatively. This split-mouth, double-blind study was performed on 24 children aged 5-7 years, who presented with bilaterally infected primary molars. Total 48 infected primary molars were divided into two groups for pulpectomy followed by obturation with ZOE and ZON, respectively. Randomization with the chit-pick method, for the control and experimental drug, was performed. A follow-up for 12 months was performed. The Chi-square test was used for inter-group comparison and the Z test was used for the analysis of data over a period of time. Overall clinical findings in this study revealed 91.7% results in ZOE compared with 100% results in the ZON group. The radiographical findings revealed 91.7% results in ZOE compared with 100% success in the ZON group. ZON demonstrated excellent effectiveness as an obturating material in the 12-month follow-up period as compared to ZOE in primary teeth. The efficacy elicited in this study could be a basis to recommend ZON as herbal alternative obturation material.