Combination of cytosine deaminase with uracil phosphoribosyl transferase leads to local and distant bystander effects against RM1 prostate cancer in mice.
We aimed to evaluate the efficacy of gene-directed enzyme-prodrug therapy (GDEPT) using cytosine deaminase in combination with uracil phosphoribosyl transferase (CDUPRT) against intraprostatic mouse androgen-refractory prostate (RM1) tumors in immunocompetent mice. The product of the fusion gene, CDUPRT, converts the prodrug, 5-fluorocytosine (5FC), into 5-fluorouracil (5FU) and other cytotoxic metabolites that kill both CDUPRT-expressing and surrounding cells, via a 'bystander effect'. Stably transformed andogen-independent mouse prostate cancer (PC) cells, RM1-CDUPRT, -GFP or GFP/LacZ cells were used. To assess the local bystander effects of CDUPRT-GDEPT, immunocompetent C57BL/6 mice implanted with cell mixtures of RM1-GFP/CDUPRT and RM1-GFP cells in different proportions intraprostatically were treated with 5FC. Pseudo-metastases in the lungs were established by a tail vein injection of untransfected RM1 cells. At necropsy, prostate weight/volume and lung colony counts were assessed. Tumors, lymph nodes, spleens and lungs were frozen or fixed for immunohistochemistry. CDUPRT expression in RM1-GFP/CDUPRT cells or tumors was confirmed by enzymic conversion of 5FC into 5FU, using HPLC. Treatment of mice bearing intraprostatic RM1-GFP/CDUPRT tumors with 5FC resulted in complete regression of the tumors. A 'local bystander effect' was seen, even though only 20% of the cells expressed CDUPRT. More importantly a significant reduction in pseudo-metastases of RM1 cells in lungs indicated a 'distant bystander effect'. Immunohistochemical evaluation of the treated tumors showed increased necrosis and apoptosis, with decreased tumor vascularity. There was also a significant increase in tumour-infiltration by macrophages, CD4+ T and natural killer cells. We conclude that CDUPRT-GDEPT significantly suppressed the aggressive growth of RM1 prostate tumors and lung pseudo-metastases via immune mechanisms involving necrosis and apoptosis.


Action potentials in epithelial taste receptor cells induced by mucosal calcium.
Chemosensory cells in the taste bud of the tongue of Necturus generate action potentials in response to electrical stimulation through a microelectrode, as recently described by Roper (Science, 220: 1311-1312, 1983). We report that the epithelial receptor cells also respond to 10 mM CaCl2, applied to the mucosal surface, with a depolarization which elicits action potentials when a threshold of -50 mV is reached. Since CaCl2 is one of the taste stimuli in amphibia, the firing of action potentials by chemoreceptor cells may be part of the signal chain in gustatory reception of Ca ions.


Rapid and quantitative method for evaluating the personal therapeutic potential of cancer drugs.
An in vitro, rapid, and quantitative cell-based assay is needed to predict the efficacy of cancer drugs in individual patients, because a cancer patient may have unconventional aspects of tumor development. Here we report a rapid and label-free quantitative method for verifying apoptosis in living cancer cells cultured on a sensor chip with a newly developed high-precision surface plasmon resonance (SPR) sensor. The time-course cell reaction was monitored as the SPR angle change rate for 5 min during a 35-min cell culture of pancreatic cancer lines with a drug. The time-course cell reaction was significantly related to cell viability counted after 48 h as assessed by caspase-3 activity assay of apoptosis. Furthermore, the detected SPR signal was derived from the decrease in inner mitochondrial membrane potential. The results obtained are universally valid for various cancer drugs mediating apoptosis through different cell-signaling pathways and even for combined use in various pancreatic cancer cell lines. This system can be applied in a clinical setting to evaluate the personal therapeutic potential of drugs including pharmacodynamic interactions.


Correlation between Colon Perfusion and Postoperative Fecal Output through a Transanal Drainage Tube during Laparoscopic Low Anterior Resection.
In order to prevent anastomotic leakage (AL) following rectal surgery, various solutions—such as intraoperative indocyanine green (ICG) angiography and transanal drainage tubes (TDT)—have been proposed. This study investigated the relationship between intestinal perfusion and fecal volume through TDT in laparoscopic low anterior resection (LAR). A total of 59 rectal cancer patients who underwent laparoscopic LAR with both intraoperative ICG angiography and postoperative TDT placement were retrospectively analyzed. The relationship between intestinal perfusion and fecal volume through TDT was examined. Based on the ICG fluorescence, the transection site was shifted more proximally in 20 cases (33.9%). Symptomatic AL occurred in seven patients (11.8%). The AL rate of the patients whose daily fecal volume exceeded 100 mL/day in 2 or more days was significantly higher than that of those whose daily fecal volume exceeded it in 0 or 1 day (44.4% vs. 6.0%; p < 0.01). Univariate and multivariate analyses showed that the need for a proximal shift of the transection site was significantly associated with a high fecal volume. The quantitative analysis of ICG fluorescence indicated that Fmax (the fluorescence difference between the baseline and maximum) was significantly associated with fecal volume through TDT.


Characterization of rat Leydig cell gonadotropin receptor structure by affinity cross-linking.
The present study was intended to examine the structure of the rat Leydig cell gonadotropin receptor. Leydig cell suspensions were prepared by either collagenase digestion or mechanical disruption of the testes. The cells were incubated with 125I-human chorionic gonadotropin (hCG) following which the bound 125I-hCG was covalently cross-linked to the cell surface receptor using a cleavable (dithiobis(succinimidyl propionate] and a noncleavable (disuccinimidyl suberate) cross-linking reagent. The extracted cross-linked membrane proteins were resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and nonreducing conditions and subjected to autoradiographic analysis. Under nonreducing conditions, three radiolabeled bands, in addition to intact hCG and its alpha-subunit, were detected with apparent molecular weights of 184,000, 136,000, and 103,000. However, under reducing conditions, three radiolabeled bands migrated on the gel corresponding to molecular weights of 144,000, 106,000, and 75,000. The binding of 125I-hCG to the receptor was inhibited by hCG and luteinizing hormone, but not by a number of other peptides or proteins. The radiolabeled bands were not detectable in hCG down-regulated Leydig cells. Furthermore, a similar autoradiographic pattern of 125I-hCG-linked complexes was seen when the 125I-linked receptor complex was subjected to immunoprecipitation with anti-hCG antibodies followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In addition, evidence was obtained indicating that these three labeled bands were derived from the same molecular species. The data suggests that the hCG receptor in Leydig cell is probably an oligomeric complex with a molecular weight of about 250,000, which is composed of three polypeptide chains of molecular weights 121,000, 83,000, and 52,000 held together through noncovalent forces. Additionally, collagenase treatment of Leydig cells does not appear to alter the autoradiographic pattern of the 125I-hCG-linked receptor.


Pollution episodes at Venice related to weather types: an analysis for a better predictability.
In order to obtain a rational strategy for the control and management of emissions from the industrial area of Venice, determination of the air quality is made from different perspectives. A source-receptor oriented approach shows a remarkable scatter of data due to the complex source field and variability of atmospheric conditions at a coastal site. As many pollution episodes are initiated by local diffusion potentials induced by the interaction between topographic irregularities (i.e. Alpine chain and Adriatic Sea) and the mesoscale atmospheric circulation system, a correlation between pollution events and synoptic conditions is made. Two periods of two years each were considered, the former characterized by very frequent, moderate episodes, the latter by more rare but potentially more severe episodes, although the emission rate was reduced. Four different criteria (i.e. weather types as classified by the U.K. Meteorological Office, by Borghi and Giuliacci, and by Urbani, and the passage of fronts) are considered and compared in order to describe the synoptic conditions and to make a reasonable prediction of the local dynamic climatology. A risk index (RI) is computed for each weather type. The U.K. classification appears to be the most suitable as it classifies the maximum number of synoptic maps and has the greatest spread of RI, thus making a more sound prediction. The analysis shows that the pollution episodes are associated with instability phenomena, i.e. passage of fronts or the onset of moderate to fresh winds, and not with stationary conditions. Also, the areal extension over which the highest pollution levels are recorded is related to particular weather types. The statistics of the meteorological weather types occurring during the pollution events give satisfactory results for some weather types, especially for those correlating highly or poorly with pollution events.


Fourteen Days vs 28 Days of Albendazole Therapy for Neurocysticercosis in Children: An Open Label Randomized Controlled Trial.
There is a paucity of literature to support 14-days albendazole therapy for neurocysticercosis (NCC). To compare the efficacy of 14-day and 28-day albendazole therapy in the management of children with newly diagnosed active NCC. Open-labelled randomized controlled trial. Children aged 1-14 years with newly diagnosed active neurocysticercosis. Albendazole (15 mg/kg/day) for either 14 days or 28 days. The primary outcome measure was proportion of children with radiological resolution of active lesion at 6-month follow up. Secondary outcome measures were proportion of children with seizure recurrence, duration to seizure recurrence and calcification on follow up imaging. 65 children with newly diagnosed NCC were rando-mized to receive albendazole therapy for 14 days (n=32) or 28 days (n=33). The proportion of children with complete resolution was comparable between the two groups [6 (18.8%) vs. 9 (27.3%); OR (95%CI):0.61 (0.19 to 1.98); P=0.56]. Similarly, proportion of children with seizure recurrence [5(15.6%) vs 2(6.1%); OR (95%CI): 2.87(0.51-16.0); P=0.26] and proportion of children with calcification on follow-up imaging [26(81.2%) vs 23(69.7%); OR (95%CI): 1.88 (0.59-5.99); P=0.39] were also comparable. There were no major side-effects noted during the study. 14-day treatment with albendazole therapy is as effective as 28-day treatment in achieving radiological resolution at six-month follow up. However, high rate of calcification in both the groups indicates need for further evaluation with an adequately powered study and longer follow up.


Inhibitor development in two patients with mild haemophilia A - spontaneous disappearance and no recurrence of the inhibitor after re-challenge.
Inhibitors against factor VIII (FVIII) complicate the treatment of patients with haemophilia. In mild haemophilia, the development of antibodies against FVIII is rare. However, the occurrence of an inhibitor in mild haemophilia changes the bleeding phenotype from mild to severe, and thus becomes a major clinical problem. We report on two patients with mild haemophilia A (FVIII level 8 and 27%, respectively), who have a missense mutation in exon 16 (G to A transition in codon 1773) and exon 22 (T to C transition in codon 2096), respectively. Both mutations have not been described in the Haemophilia A Mutation, Structure Test and Resource Site. Our patients developed high titer inhibitors following an intensive FVIII replacement therapy due to a muscle bleeding and after a polytrauma. During the presence of the inhibitor, AICC or FVIIa was successfully used as bypassing agent. In both patients the inhibitor disappeared spontaneously. Years after the development of the inhibitor, the patients again received FVIII concentrates. Reappearance of the inhibitor was not observed in either patient. The reported cases indicate that inhibitors in patients with mild haemophilia might be transient and disappear spontaneously. Therefore, the necessity of immune tolerance therapy, which is costly and strenuous for the patients, should be critically examined for each individual patient and a watch and wait strategy might be advisable.


Primary periareolar abscess in the nonlactating breast: risk of recurrence.
The estimated probability of relapse of primary periareolar abscess of the nonlactating breast was 38 percent (95 percent confidence interval 18 to 58 percent) 8 years after simple incision and healing by granulation. Relapses were mainly fistulas in the line of incision. An attempt to identify relapses early was unsuccessful. This finding, in combination with the unsatisfactory results of simple incision and healing by granulation and the character of relapses, suggests improper initial operation and multifactorial causes. Initial incision followed by a search for a possible sinus tract and extensive excision in all patients with primary abscesses has been described and is recommended.


Compensatory strategies during manual wheelchair propulsion in response to weakness in individual muscle groups: A simulation study.
The considerable physical demand placed on the upper extremity during manual wheelchair propulsion is distributed among individual muscles. The strategy used to distribute the workload is likely influenced by the relative force-generating capacities of individual muscles, and some strategies may be associated with a higher injury risk than others. The objective of this study was to use forward dynamics simulations of manual wheelchair propulsion to identify compensatory strategies that can be used to overcome weakness in individual muscle groups and identify specific strategies that may increase injury risk. Identifying these strategies can provide rationale for the design of targeted rehabilitation programs aimed at preventing the development of pain and injury in manual wheelchair users. Muscle-actuated forward dynamics simulations of manual wheelchair propulsion were analyzed to identify compensatory strategies in response to individual muscle group weakness using individual muscle mechanical power and stress as measures of upper extremity demand. The simulation analyses found the upper extremity to be robust to weakness in any single muscle group as the remaining groups were able to compensate and restore normal propulsion mechanics. The rotator cuff muscles experienced relatively high muscle stress levels and exhibited compensatory relationships with the deltoid muscles. These results underline the importance of strengthening the rotator cuff muscles and supporting muscles whose contributions do not increase the potential for impingement (i.e., the thoracohumeral depressors) and minimize the risk of upper extremity injury in manual wheelchair users.


Causes of anaemia in very low birth weight infants. Phlebotomy losses are not the first accused.
The specific aim of the study was to determine the correlation between the severity of pathology, the amount of blood removed for diagnostic purposes in the 1st week of life and the incidence of early anaemia in very low birth weight (VLBW) infants. We recorded the level of haemoglobin (Hb) and haematocrit (Ht) in each of the 50 infants entered in the study at their admission in our neonatal intensive care unit (NICU) and at the age of 8 days. We quantified for each infant the blood drawn for clinical purpose during the 1(st)week of life, using microanalytic techniques for all types of analysis performed. Using the neonatal therapeutic intensive score system (NTISS) we divided all patients into 2 groups: group A= mild light pathology; group B= severe pathology. There was statistically significant difference between the percent decrease of Hb and Ht with reference to the birth weight in the 2 groups. Logistic regression analysis indicated a strong correlation (P = 0.009) between higher degree of illness severity and higher percent decrease of Hb and Ht. The difference due to the amount of phlebotomy losses was not significant. To our knowledge, this study is the first that strongly suggest that phlebotomy losses is not the main cause of anaemia in VLBW preterm infants in the 1st week of life, when a policy of strictly attention to the amount of blood removed is performed.


Functional correction of established central nervous system deficits in an animal model of lysosomal storage disease with feline immunodeficiency virus-based vectors.
Gene transfer vectors based on lentiviruses can transduce terminally differentiated cells in the brain; however, their ability to reverse established behavioral deficits in animal models of neurodegeneration has not previously been tested. When recombinant feline immunodeficiency virus (FIV)-based vectors expressing beta-glucuronidase were unilaterally injected into the striatum of adult beta-glucuronidase deficient [mucopolysaccharidosis type VII (MPS VII)] mice, an animal model of lysosomal storage disease, there was bihemispheric correction of the characteristic cellular pathology. Moreover, after the injection of FIV-based vectors expressing beta-glucuronidase into brains of beta-glucuronidase-deficient mice with established impairments in spatial learning and memory, there was dramatic recovery of behavioral function. Cognitive improvement resulting from expression of beta-glucuronidase was associated with alteration in expression of genes associated with neuronal plasticity. These data suggest that enzyme replacement to the MPS VII central nervous system goes beyond restoration of beta-glucuronidase activity in the lysosome, and imparts improvements in plasticity and spatial learning.


Association between inflammatory mediators and muscle mass in long-term hemodialysis patients.
Muscle wasting is highly prevalent in long-term hemodialysis (HD) patients. Although inflammatory indices have been associated with malnutrition in these patients, the role of inflammation in muscle wasting has not yet been determined. The relationship between the inflammatory mediators C-reactive protein (CRP) and interleukin-6 (IL-6) and the muscle mass indices thigh muscle area (TMA), measured by computed tomography, and creatinine (Cr) production, estimated by the Cr kinetic model (Cr-CKM), were investigated in 188 HD patients. Serum IL-6 level (7.3 +/- 7.8 pg/mL) was significantly elevated in HD patients, whereas mean serum CRP level (4.8 +/- 7.5 mg/L) remained within the normal range. Similar to serum albumin, muscle mass indices had significantly negative correlations with both serum IL-6 and CRP levels (TMA/dry weight [DW] versus log IL-6, r = -0.28; P < 0.01; TMA/DW versus log CRP, r = -0.38; P < 0.001; Cr-CKM versus log IL-6, r = -0.31; P < 0.01; Cr-CKM versus log CRP, r = -0.24; P < 0.01). Although muscle mass indices also were associated with both age and sex, a multiple regression analysis confirmed that these inflammatory indices were significantly associated with muscle mass in HD patients. Data indicate that muscle wasting is associated closely with inflammatory indices in long-term HD patients. It may be important to clarify the mechanism for the increasing inflammatory status and suppress the inflammatory response in these patients to improve their malnutrition and recover muscle mass.


Influence of effector molecules on the CD8(+) T cell response to infection.
Expansion and contraction of antigen-specific CD8(+) T cells after infection or vaccination results in stable memory. Recent evidence demonstrates that anti-microbial effector molecules such as perforin and IFN-gamma regulate specific aspects of CD8(+) T cell homeostasis by mechanisms that may be dependent or independent of pathogen clearance.


An interferon gamma response signature links myocardial aging and immunosenescence.
Aging entails profound immunological transformations that can impact myocardial homeostasis and predispose to heart failure. However, preclinical research in the immune-cardiology field is mostly conducted in young healthy animals, which potentially weakens its translational relevance. Herein, we sought to investigate how the aging T-cell compartment associates with changes in myocardial cell biology in aged mice. We phenotyped the antigen-experienced effector/memory T cells purified from heart-draining lymph nodes of 2-, 6-, 12-, and 18-month-old C57BL/6J mice using single-cell RNA/T cell receptor sequencing. Simultaneously, we profiled all non-cardiomyocyte cell subsets purified from 2- to 18-month-old hearts and integrated our data with publicly available cardiomyocyte single-cell sequencing datasets. Some of these findings were confirmed at the protein level by flow cytometry. With aging, the heart-draining lymph node and myocardial T cells underwent clonal expansion and exhibited an up-regulated pro-inflammatory transcription signature, marked by an increased interferon-γ (IFN-γ) production. In parallel, all major myocardial cell populations showed increased IFN-γ responsive signature with aging. In the aged cardiomyocytes, a stronger IFN-γ response signature was paralleled by the dampening of expression levels of transcripts related to most metabolic pathways, especially oxidative phosphorylation. Likewise, induced pluripotent stem cells-derived cardiomyocytes exposed to chronic, low grade IFN-γ treatment showed a similar inhibition of metabolic activity. By investigating the paired age-related alterations in the T cells found in the heart and its draining lymph nodes, we provide evidence for increased myocardial IFN-γ signaling with age, which is associated with inflammatory and metabolic shifts typically seen in heart failure.


New fluorogenic dansyl-containing calix[4]arene in the partial cone conformation for highly sensitive and selective recognition of lead(II).
A new efficient and highly selective fluorescent chemosensor for determination of Pb (2+) has been obtained by covalent attachment of two pendent proton-ionizable dansylcarboxamide groups to the calix[4]arene preorganized in the partial cone conformation. This geometry of the calixarene moiety was chosen on the basis of the prior (1)H NMR study of conformations adopted by the flexible dansyl-containing prototype upon complexation with lead ion. In acidic MeCN-H2O (1:1 v/v) solutions, the partial cone fluoroionophore allowed for detection of Pb (2+) at the levels as low as 2.5 ppb, which is totally compatible with the regulations of the U.S. Environmental Protection Agency and the World Health Organization on the limiting content of this hazardous pollutant in drinking water.


Quality control of photosystem II under light stress - turnover of aggregates of the D1 protein in vivo.
When photodamaged under excessive light, the D1 protein is digested and removed from Photosystem (PS) II to facilitate turnover of the protein. In vitro studies have shown that part of the photodamaged D1 protein forms aggregates with surrounding polypeptides before being digested by a protease(s) in the stroma [Yamamoto Y (2001) Plant Cell Physiol 42: 121-128]. The aim of this study was to examine whether light-induced aggregation of the D1 protein also occurs in vivo. The following results were obtained: (1) PS II activity in spinach leaves was significantly inhibited by weak illumination (light intensity, 20-100 microE m-2 s-1), as monitored by chlorophyll fluorescence Fv/Fm, when the leaves were kept at higher temperatures (35-40 degrees C); (2) aggregation of the D1 protein, as well as cleavage of the protein, was detected in thylakoids isolated from spinach leaves that had been subjected to heat/light stress; (3) aggregates of the D1 protein disappeared after incubation of the leaves at 25 degrees C in the dark or under illumination with weak light. Since it is dependent on the presence of oxygen, aggregation of the D1 protein is probably induced by reactive oxygen species produced in thylakoids upon illumination at elevated temperatures. Consistent with this notion, singlet oxygen production in thylakoid samples under illumination was shown to be stimulated significantly at higher temperatures.


Targeted re-sequencing in malformations of cortical development: genotype-phenotype correlations.
Malformations of cortical development (MCD) are a phenotypically and genetically heterogeneous group of disorders, for which the diagnostic rate of genetic testing in a clinical setting remains to be clarified. In this study we aimed to assess the diagnostic rate of germline and pathogenic variants using a custom panel in a heterogeneous group of subjects with MCD and explore genotype-phenotype correlations. A total of 84 subjects with different MCD were enrolled. Genomic DNA was isolated from peripheral blood. Fifty-nine tartget genes were assessed using a custom next-generation sequencing (NGS) panel. Genetic causes were identified in one-fourth of our cohort (21.4 %). Overall, we identified 19 pathogenic or likely pathogenic single-nucleotide variants in 11 genes among 18 subjects, including PAFAH1B1 (LIS1) (n = 3), TUBA1A (n = 3), DYNC1H1 (n = 3), ACTG1 (n = 2), TUBB2B (n = 1), TUBB3 (n = 1), DCX (n = 1), FLNA (n = 1), LAMA2 (n = 1), POMGNT2 (n = 1) and VLDLR (n = 1). The diagnostic yield was higher in patients with lissencephaly/pachygyria (60 %) (p = 0.001), cobblestone malformation (50 %), and subcortical band heterotopia (SBH) (40 %). Furthermore, five out of six subjects with suspect tubulinopathies on imaging harboured pathogenic variants in tubulin genes. Overall, germline pathogenic variants were more likely to be identified if MCD were diffuse (p = 0.002) and associated with other central nervous system malformations (p = 0.029). Moderate to severe intellectual disability was also more commonly associated with pathogenic variants (p = 0.044). Customized gene panels may support the diagnostic work-up for some specific MCD, especially when these are diffuse, bilateral and associated with other brain malformations.


Characteristics of Optic Disc Rotation in Myopic Eyes.
To investigate the characteristics of optic disc rotation and ocular parameters affecting optic disc rotation in healthy myopic eyes. Cross-sectional, comparative study. A total of 220 participants with healthy myopic eyes. Spherical equivalent (SE) refractive error, axial length, central corneal thickness, and intraocular pressure (IOP) were evaluated. Optic disc tilt ratio, degree of optic disc rotation, and area of β-zone parapapillary atrophy (PPA) were measured. Optic nerve head (ONH) parameters and thickness of the peripapillary retinal nerve fiber layer (pRNFL) and macular ganglion cell-inner plexiform layer (mGCIPL) were measured using Cirrus optical coherence tomography (Carl Zeiss Meditec Inc., Dublin, CA). Subjects were divided into 2 groups, group 1 with superior rotation and group 2 with inferior rotation of the optic disc, and various parameters were compared. Linear regression analysis was performed to evaluate the relationships between the degree of optic disc rotation and several parameters. Degree of optic disc rotation. Among 220 eyes, 147 showed superior rotation of the optic disc and 73 showed inferior rotation. The mean tilt ratio and rotation degree were 1.16 and -19.51°, respectively, in group 1 and 1.20 and 28.93°, respectively, in group 2, showing significant differences between the groups (P = 0.028 and P = 0.035, respectively). There were also significant between-group differences in IOP (15.59 vs. 16.34 mmHg), SE refractive error (-4.05 vs. -5.66 diopters [D]), axial length (25.51 vs. 26.26 mm), and area of β-zone PPA (0.32 vs. 0.70 mm(2)). Overall, a multivariate linear regression analysis showed that IOP, axial length, and area of β-zone PPA were significant parameters related to the degree of optic disc rotation (P = 0.011, P = 0.043, and P = 0.030, respectively). Group 2 showed thinner pRNFL and mGCIPL thickness in general compared with group 1. In healthy myopic eyes, superior rotation of the optic disc was more prevalent than inferior rotation. As the optic disc rotates inferiorly, there was a significant positive correlation with IOP, axial length, and area of the β-zone PPA. Conversely, a significant negative correlation with pRNFL and mGCIPL thickness was observed.


Transatrial Endocardial Placement of Epicardial Pacing Lead.
We report a technique wherein an epicardial pacing lead was placed transatrially to achieve optimal pacing in a patient with a complex venous anatomy.


The temporomandibular joint pain dysfunction syndrome.
This paper discusses a relatively common myofascial pain syndrome. Despite its commonness, it is frequently misdiagnosed and patients may be subjected to many forms of treatment without relief. Some of these treatments may be mutilating, or may produce complications which add to the patient's discomfort. It is therefore suggested that competent dental advice should be sought in all cases of atypical facial pain, and that pain clinics should include a dentist as a member of the team.


Sudden infant death in Copenhagen 1956--1971. II. Social factors and morbidity.
131 cases of the Sudden Infant Death Syndrome (SIDS) among infants born in the Municipality of Copenhagen during 1956--1971 were analysed on the basis of data collected prospectively by the infant health visitors and abstracted from police reports. Compared with controls, a significantly larger number of SIDS infants had been born out of wedlock, were living only with their mother, had parents in a less secure occupation, lived in more crowded, poor-quality dwellings and districts. The "home standard" was lower among the SIDS families, including a lower standard of infant care, a higher percentage of not keeping the appointments with the infant health visitors and a lower mental capacity in the mothers; there was no difference with respect to the mother's physical capacity. More congenital malformations and more cases of asphyxia were found among the SIDS infants, and a significantly larger number of the SIDS infants had been admitted to hospital, with a tendency to being kept there longer. It is concluded that a relationships exists between poor social conditions, increased morbidity, and SIDS. At the same time, it is pointed out that during recent years the differences between cases and controls as regards certain of the social parameters studied have decreased.


Increased glomerular atrial natriuretic peptide receptor affinity in experimental nephrotic syndrome.
Atrial natriuretic peptide (ANP) is a cardiac hormone with natriuretic and diuretic effects. To better define the ANP hormonal system in the nephrotic syndrome, a condition associated with renal sodium retention, we undertook a study of glomerular ANP receptors in rats with puromycin aminonucleoside-induced nephrotic syndrome and in pair-fed controls. Nephrotic rats had significantly decreased serum albumin and total protein and significantly increased serum cholesterol, triglycerides and 24 hour urinary protein excretion. Plasma level of atrial natriuretic peptide was similar in both groups of rats. Competition binding inhibition studies in isolated glomeruli demonstrated one binding site in both groups of rats. The density of ANP binding sites in isolated glomeruli was similar in nephrotic and pair-fed rats while the binding affinity was increased significantly in the nephrotic rats. This is the first study to demonstrate alterations in renal ANP receptors in the nephrotic syndrome. Further studies will be necessary to determine whether alterations in glomerular ANP receptors contribute to renal sodium retention in the nephrotic syndrome.


Changes in paroxetine binding in the cerebral cortex of polydipsic rats.
Schedule-induced polydipsia was induced when food-deprived rats were subjected to a fixed-time (60 s) feeding schedule for 150 min daily for 3 weeks (training period). Subsequent chronic administration of the serotonin reuptake inhibitor fluoxetine reduces schedule-induced polydipsia over 2-4 weeks. We asked whether changes in the serotonin reuptake carrier occur following the development of schedule-induced polydipsia and its reduction by fluoxetine. Using [3H]paroxetine binding, we found a 40% increase in Kd and a 50% decrease in Bmax in polydipsic rats; both were reversed by fluoxetine. Food deprivation alone did not affect these parameters. These observations suggest that changes in the serotonin reuptake carrier correlate with the development and reversal of schedule-induced polydipsia.


Calculation of odour emissions from aircraft engines at Copenhagen Airport.
In a new approach the odour emissions from aircraft engines at Copenhagen Airport are calculated using actual fuel flow and emission measurements (one main engine and one APU: Auxiliary Power Unit), odour panel results, engine specific data and aircraft operational data for seven busy days. The calculation principle assumes a linear relation between odour and HC emissions. Using a digitalisation of the aircraft movements in the airport area, the results are depicted on grid maps, clearly reflecting aircraft operational statistics as single flights or total activity during a whole day. The results clearly reflect the short-term temporal fluctuations of the emissions of odour (and exhaust gases). Aircraft operating at low engine thrust (taxiing, queuing and landing) have a total odour emission share of almost 98%, whereas the shares for the take off/climb out phases (2%) and APU usage (0.5%) are only marginal. In most hours of the day, the largest odour emissions occur, when the total amount of fuel burned during idle is high. However, significantly higher HC emissions for one specific engine cause considerable amounts of odour emissions during limited time periods. The experimentally derived odour emission factor of 57 OU/mg HC is within the range of 23 and 110 OU/mg HC used in other airport odour studies. The distribution of odour emission results between aircraft operational phases also correspond very well with the results for these other studies. The present study uses measurement data for a representative engine. However, the uncertainties become large when the experimental data is used to estimate the odour emissions for all aircraft engines. More experimental data is needed to increase inventory accuracy, and in terms of completeness it is recommended to make odour emission estimates also for engine start and the fuelling of aircraft at Copenhagen Airport in the future.


Surgical attenuation of spontaneous congenital portosystemic shunts in dogs resolves hepatic encephalopathy but not hypermanganesemia.
Hypermanganesemia is commonly recognized in human patients with hepatic insufficiency and portosystemic shunting. Since manganese is neurotoxic, increases in brain manganese concentrations have been implicated in the development of hepatic encephalopathy although a direct causative role has yet to be demonstrated. Evaluate manganese concentrations in dogs with a naturally occurring congenital shunt before and after attenuation as well as longitudinally following the changes in hepatic encephalopathy grade. Our study demonstrated that attenuation of the shunt resolved encephalopathy, significantly reduced postprandial bile acids, yet a hypermanganasemic state persisted. This study demonstrates that resolution of hepatic encephalopathy can occur without the correction of hypermanganesemia, indicating that increased manganese concentrations alone do not play a causative role in encephalopathy. Our study further demonstrates the value of the canine congenital portosystemic shunt as a naturally occurring spontaneous model of human hepatic encephalopathy.


Singapore's diverse immunology landscape: interconnected research networks enabling blue sky ideas through to translation.
Singapore stands as a dynamic hub for cutting-edge immunological research and innovation. The country's vibrant research ecosystem is supported by collaborative networks across the many national medical and scientific research institutes, fostering meaningful alliances between academia and industry. In this article, we speak to Assistant Professor Jinmiao Chen from the Agency for Science, Technology, and Research (A*STAR) and Professor Nicholas Gascoigne from the National University of Singapore (NUS), Duke-NUS Medical School and Nanyang Technological University (NTU) about immunology in Singapore. Credit: Kate Forbes.


An exploratory experimental study examining the short-term effects of a public service announcement on parents' perceptions of the descriptive norm for children's outdoor play: A cautionary tale.
Public service announcements (PSA) targeting physical activity are not always effective at enacting behaviour change. One factor that may impact message effectiveness is whether the message matches its intent (ie, what norm is being portrayed and received). The current study explored the effects of two PSAs on parents' perceptions of outdoor play. Parents (N = 333) completed a pre/postexperimental online survey to examine the acute impact of a PSA on their perception of how often children play outdoors (ie, descriptive norms). Participants provided their descriptive norm perceptions and then were randomised to view one of two PSAs: (a) the Mr. Lonely PSA, which portrayed outdoor activity as not normative, and (b) a control message, which portrayed outdoor activity as the norm (VERB PSA). After viewing their PSA, descriptive norm perceptions were reported again. A two-way mixed ANOVA indicated that participants who viewed the Mr. Lonely PSA (where outdoor play was not normative) perceived that children and youth played outdoors less frequently immediately after watching the PSA compared to before viewing it (P < 0.05). No significant change occurred in the control condition. PSAs can alter individuals' perceptions of whether they perceive outdoor play as normative among children and youth. SO WHAT?: As individuals align their behaviour with what is typical, PSA developers should be wary of what message is being conveyed. In terms of activity, portraying that outdoor play is not normative may not have the intended outcome.


Psychology--tensions between objectivity and subjectivity.
The self is often defined in terms of its presentational appearances. This may easily end up in a denial of the internal aspects of the self, which is very often related to a tendency to avoid the tension between the internal and the external, but also between subjectivity and objectivity. In this paper this ambition is regarded in a historical perspective, in which Fichte and Hegel both represent attempts at abolishing the tension, whereas Kant and Kierkegaard represent the opposite. History shows that an eradication of the tension between subjectivity and objectivity implies a deterioration of psychology as well. Thus the conclusion is that psychology is primarily to be defined in terms of the tension between subjectivity and objectivity, which requires an accurate understanding and the inclusion of both of them.


Acta Anatomica: A portrait of an anatomy department, Christmas 1951.
Seventy years ago, two medical art students painted a group portrait of the staff of the anatomy department in the University of Manchester Medical School. The painting is an unusual allegorical portrayal of the staff as pantomime characters. This paper asks: who were they and what were their subsequent careers? Does this picture tell us anything about the role of anatomy in medical education in the 1950s?


Children with Complex Mental Health Problems: Needs, Costs and Predictors over One Year.
Little is known regarding children of greatest concern with complex mental health problems. A one-year prospective study of psychiatric diagnosis, psychosocial functioning, need status and service receipt in 60 children identified as most concerning. Thirty-two (53%) had two or more disorders. The mean number of needs per child was five. One year later mean needs were unchanged but with considerable individual variation. Mean weekly costs were £1017 (€1627) (SD = £957 (€1531)). Higher costs related to social factors rather than diagnosis or need. The mean annual cost of services to children with complex mental health problems is ten times that in other studies of children with mental health problems.


Isolation and sequence analysis of cDNA clones coding for rat skeletal muscle creatine kinase.
A series of cDNA clones corresponding to 1494 bases of rat muscle creatine kinase mRNA has been isolated and characterized. The identity of these clones has been confirmed by DNA sequence analysis and by comparison of the predicted amino acid sequence with that determined for the purified protein. The cDNA sequence accounts for the entire coding sequence of the creatine kinase protein in addition to the complete 3' untranslated region and 68 bases of 5' noncoding region. Sequences corresponding to the active site region of the protein, the initiation codon, the termination codon, and poly(A) addition signal have been identified.


Inclusion as a focus of employment-related research in intellectual disability from 2000 to 2010: a scoping review.
This paper presents the results of a scoping review of the research literature on community-based employment for individuals with intellectual disabilities from 2000 to 2010. The review examined the variables studied in each paper, and considered the degree to which elements of social inclusion were addressed. The search strategy identified a total of 245 articles, the total pool of which was reduced to 42 following abstract and text review. Two researchers reviewed the final set of articles and extracted information relevant to the study goals. Independent and dependent measures used in the studies were categorized relative to a conceptual model of social inclusion. The frequency with which each aspect of inclusion was addressed in the studies was totalled, and the resulting pattern analyzed qualitatively. The analysis revealed that the majority of papers identified the work role achieved (i.e. employment and pay rates, job titles) as the primary construct of interest, while fewer than 5 articles focused on central aspects of inclusion, such as sense of belonging, reciprocity, and need fulfillment. This study profiles the evidence base relative to inclusive employment for people with intellectual disabilities. The lack of evidence on the degree to which social inclusion is being achieved through community-based employment highlights a critical area requiring attention.


[Central pseudocancerous form of bronchopulmonary tuberculosis simulating cancer].
Pathomorphism of tuberculosis results in an increased frequency of forms simulating bronchial carcinoma. A specific bronchopulmonary lesion involving atelectasis of the upper lobe is reported. Since it occurs in middle-aged female patients and develops mostly in the left lung, the lesion should be regarded as a specific pseudotumorous form of pulmonary tuberculosis.


Evaluation of esophageal contractile propagation using esophageal pressure topography.
High-resolution manometry and esophageal pressure topography have enhanced our ability to analyze esophageal motor disturbances by improving the detail and accuracy of measurements of peristaltic activity.This has been extremely helpful in the evaluation of disorders of rapid propagation as the technique is able to define important time points and physiologic landmarks that are crucial in defining peristaltic velocity and latency intervals. The goal of the current review will be to assess how esophageal pressure topography has impacted our ability to define important phenotypes of rapid propagation. Additionally, this review will also be utilized to complement the description of the Chicago Classification of Esophageal Motor Disorders, which is presented in this supplement issue.


Blood eosinophils and eosinophil cationic protein after acute myocardial infarction or corticosteroid administration.
Measurements of eosinophil cationic protein in serum (S-ECP) have been made in patients with acute myocardial infarction. In spite of low numbers of blood eosinophils in the acute phase of the disease S-ECP levels fluctuated widely with often markedly raised levels. Peak levels of S-ECP were significantly correlated (P less than 0.001) to peak numbers of blood eosinophils, although the former usually occurred within the first 2-3 d of illness and the latter usually after the sixth day. Intravenous injection of 0.5 g methylprednisolone in healthy individuals reduced blood eosinophil counts for at least 24 h, but S-ECP levels remained within normal limits. These results suggest that ECP is released from eosinophils in vivo. The raised levels of S-ECP in patients with acute myocardial infarction are probably due to the active participation of eosinophils in the inflammatory process.


Updated immunomodulatory roles of gut flora and microRNAs in inflammatory bowel diseases.
Inflammatory bowel disease is a heterogeneous intestinal inflammatory disorder, including ulcerative colitis (UC) and Crohn's disease (CD). Existing studies have shown that the pathogenesis of IBD is closely related to the host's genetic susceptibility, intestinal flora disturbance and mucosal immune abnormalities, etc. It is generally believed that there are complicated interactions between host immunity and intestinal microflora/microRNAs during the occurrence and progression of IBD. Intestinal flora is mainly composed of bacteria, fungi, viruses and helminths. These commensals are highly implicated in the maintenance of intestinal microenvironment homeostasis alone or in combination. MiRNA is an endogenous non-coding small RNA with a length of 20 to 22 nucleotides, which can perform a variety of biological functions by silencing or activating target genes through complementary pairing bonds. A large quantity of miRNAs are involved in intestinal inflammation, mucosal barrier integrity, autophagy, vesicle transportation and other small RNA alterations in IBD circumstance. In this review, the immunomodulatory roles of gut flora and microRNAs are updated in the occurrence and progression of IBD. Meanwhile, the gut flora and microRNA targeted therapeutic strategies as well as other immunomodulatory approaches including TNF-α monoclonal antibodies are also emphasized in the treatment of IBD.


Bilateral inter-arm coordination in freestyle swimming: effect of skill level and swimming speed.
The aim of this study was to examine the influence of level of skill and swimming speed on inter-limb coordination of freestyle swimming movements. Five elite (2 males, 3 females; age 18.9+/-1.0 years, height 1.71+/-0.04 m, body mass 62.1+/-7.0 kg) and seven novice (age 22.0+/-2.0 years, height 1.77+/-0.04 m, body mass 74.8+/-9.0 kg) swimmers swam a sprint and a self-paced 25 m freestyle trial. The swimming trials were recorded by four digital cameras operating at 50 Hz. The digitized frames underwent a three-dimensional direct linear transformation to yield the three-dimensional endpoint kinematic trajectories. The spatio-temporal relationship between the upper limbs was quantified by means of the peak amplitude and time lag of the cross-correlation function between the right and left arm's endpoint trajectories. A strong anti-phase coupling between the two arms, as confirmed by peak amplitudes greater than 0.8, was noted for both groups and swimming speeds. Significantly higher (P<0.05) peak amplitudes were observed for the sprint compared with self-paced swimming. No significant differences in the strength of inter-limb coupling were noted between the elite and novice swimmers (P>0.05). Time lags were very close to 0 ms and did not differ between groups or swimming speeds. We conclude that in freestyle swimming, the intrinsic anti-phase (180 degrees phase difference) inter-limb relationship is strongly preserved despite the physically powerful environmental influence of the water and this "preferred" pattern is not affected by level of skill. In contrast, increasing movement speed results in stronger inter-limb coupling that is closer to the anti-phase inter-limb relationship.


Heterogeneity of immunoreactive prolactin in the rat brain.
Three immunoreactive prolactin proteins (24 Kd, 16 Kd, and 12 Kd) were identified in the rat brain using sodium dodecyl sulphate polyacrylamide gel electrophoresis, and western blot analyses. In male and female brains, the primary prolactin protein has a molecular weight of 24 Kd which is similar to that of pituitary prolactin. Two additional proteins with apparent molecular weights of 16 Kd and 12 Kd were also identified and were found in greater concentrations in the brain than in the pituitary, and were more predominant in the female brain. In addition, brain extracts proteolytically modify the 24K dalton PRL resulting in the formation of two fragments with apparent molecular weights of 16 and 8 Kd. These data indicate that the prolactin identified in the rat brain is similar to pituitary prolactin, and suggests, that like other PRL target tissues the brain may have the capacity to proteolytically modify prolactin.


Gas Diffusion Layer with a Regular Hydrophilic Structure Boosts the Power Density of Proton Exchange Membrane Fuel Cells via the Construction of Water Highways.
The gas diffusion layer (GDL) is an essential carrier for the mass transmission of proton exchange membrane fuel cells (PEMFCs), which decides the peak power density of PEMFCs. Herein, a gas diffusion layer with a regularly arranged hydrophilic and hydrophobic pattern structure was prepared by a template method combined with the ultrasonic spray process. The peak power density was enhanced by 30% (from 520 to 678 mW/cm2) compared to an unpatterned structure, and the breakthrough pressure of the GDL was reduced from 13.61 to 2.96 kPa. In addition, the finite element analysis (FEA) results indicate that the polarization curve of calculation was highly consistent with the experimental results. Importantly, the capillary pressure of the hydrophilic area was about 0.3 kPa, much lower than that of the hydrophobic area (2 kPa), demonstrating that the hydrophilic and hydrophobic synergistic structure reduced the water transmission resistance in separating water and oxygen and builds a high-speed channel for water transmission.


β1-adrenergic receptor and sphingosine-1-phosphate receptor 1 (S1PR1) reciprocal downregulation influences cardiac hypertrophic response and progression to heart failure: protective role of S1PR1 cardiac gene therapy.
The sphingosine-1-phosphate receptor 1 (S1PR1) and β1-adrenergic receptor (β1AR) are G-protein-coupled receptors expressed in the heart. These 2 receptors have opposing actions on adenylyl cyclase because of differential G-protein coupling. Importantly, both of these receptors can be regulated by the actions of G-protein-coupled receptor kinase-2, which triggers desensitization and downregulation processes. Although classic signaling paradigms suggest that simultaneous activation of β1ARs and S1PR1s in a myocyte would simply result in opposing action on cAMP production, in this report we have uncovered a direct interaction between these 2 receptors, with regulatory involvement of G-protein-coupled receptor kinase-2. In HEK (human embryonic kidney) 293 cells overexpressing both β1AR and S1PR1, we demonstrated that β1AR downregulation can occur after stimulation with sphingosine-1-phosphate (an S1PR1 agonist), whereas S1PR1 downregulation can be triggered by isoproterenol (a β-adrenergic receptor agonist) treatment. This cross talk between these 2 distinct G-protein-coupled receptors appears to have physiological significance, because they interact and show reciprocal regulation in mouse hearts undergoing chronic β-adrenergic receptor stimulation and in a rat model of postischemic heart failure. We demonstrate that restoration of cardiac plasma membrane levels of S1PR1 produces beneficial effects that counterbalance the deleterious β1AR overstimulation in heart failure.


Correction for under-ascertainment in cancer cases in the very elderly (aged 75+): external reference method.
Any fall-off in the age-specific cancer incidence curve in oldest age groups may be suggestive of under-ascertainment. This article introduces a correction method for under-ascertainment of cancer cases in the very elderly (aged 75+). Corrected age-specific rates for a certain cancer are calculated by summing the age-specific rates of that cancer for two lower age groups multiplied by a corresponding coefficient from an external reference cancer registry (sex-, site-, and age-specific coefficients from the Finnish Cancer Registry, a nation-wide population-based registry with high data validity). In order to examine this new method, Lithuanian Cancer Registry data for the period 1993-97 was chosen as an example. Age-specific incidence curves of cancers in both sexes and most of cancer sites in Lithuania demonstrated the presence of under-ascertainment in the oldest age groups (over 75). Comparison of the corrected incidence rates for 1995 with the original ones showed that without correction for under-ascertainment, crude rates (and consequently number of new cases) were underestimated by around 10%. Underestimation for the year 2050 in Lithuania was approximately 20%. Depending on cancer site and sex, the percentage of underestimation varied substantially. In many cancer registries, after correction for under-ascertainment in elderly age groups, the estimates of number of new cases and incidence rates increase. With respect to prepare future projections of incidence, when the effects of an aging population need to be taken into account, accurate age-specific rates in the upper age groups are important. Without correction, cancer occurrence measures may be substantially underestimated, which may lead to inadequate resource allocations for cancer control.


Association of Hypertriglyceridemic Waist Phenotype with Obstructive Sleep Apnea: A Cross-Sectional Study.
Hypertriglyceridemic waist (HTGW) phenotype is an independent risk factor for metabolic disorders. Although obstructive sleep apnea (OSA) is associated with metabolic disorders, it is unclear whether there is an association between HTGW phenotype and OSA. We enrolled consecutive participants presenting to a sleep center in Shanghai, China. Full-night polysomnography was performed, and serum triglyceride (TG) levels and waist circumference (WC) were calculated. HTGW phenotype was defined as increased WC (men > 90 cm, women > 80 cm) and elevated TG levels (> 1.7 mmol/L). Participants were classified into four groups: normal TG with normal WC (NTNW); normal TG with increased WC (NTGW); elevated TG with normal WC (HTNW); and elevated TG with enlarged WC, namely HTGW. The relationships between HTGW phenotypes and OSA were assessed using binary (apnea hypopnea index, [AHI]≥5/hr) and multinomial logistic regression analyses (clinical AHI severity categories). We included 3190 participants in this cross-sectional study. Compared to the NTNW phenotype, participants with NTGW and HTGW phenotypes had the significantly higher risk of OSA (AHI ≥5/hr, odds ratio [OR] = 2.51, 95% confidence interval [CI] = 1.91-3.31; OR = 3.76, 95% CI = 2.67-5.31, respectively), after adjustment for confounders. In subgroup analyses categorised by age, sex, and BMI, the aforementioned associations remained significant. The association between the NTGW, HTGW phenotype and OSA risk remained significant across the OSA severity groups. Multinomial logistic regression also revealed that the mild (OR = 1.63, 95% CI = 1.07-2.50), moderate (OR = 1.875, 95% CI = 1.22-2.88), and severe OSA (OR = 3.18, 95% CI = 2.14-4.73) were associated with HTGW phenotype. Both NTGW and HTGW phenotype were positively associated with OSA risk in all subgroups. Further longitudinal follow-up studies are needed to determine the causal link and prognostic role of these metabolic factors.


Specification of vertebrate slow-twitch muscle fiber fate by the transcriptional regulator Blimp1.
Skeletal muscles of vertebrates are typically composed of slow- and fast-twitch fibers that differ in their morphology, gene expression profiles, contraction speeds, metabolic properties and patterns of innervation. During myogenesis, how muscle precursors are induced to mature into distinct slow- or fast-twitch fiber-types is inadequately understood. We have previously shown that within the somites of the zebrafish embryo, the activity of the zinc finger and SET domain-containing transcriptional regulator Blimp1 is essential for the specification of slow muscle fibers. Here, we have investigated the mechanism by which Blimp1 programs myoblasts to adopt the slow-twitch fiber fate. In slow myoblasts, expression of the Blimp1 protein is transient, and precedes the expression of slow muscle-specific differentiation genes. We demonstrate that the competence of somitic myoblasts to commit to the slow lineage in response to Blimp1 changes as a function of developmental time. Furthermore, we provide evidence that mammalian Blimp1 can recapitulate the slow myogenic program in zebrafish, suggesting that zebrafish Blimp1 can recognize the same consensus DNA sequence that is bound by the mammalian protein. Finally, we show that zebrafish Blimp1 can repress the expression of fast muscle-specific myosin light chain, mylz2, through direct binding near the promoter of this gene, indicating that an important function of the transcriptional activity of Blimp1 in slow muscle development is the suppression of fast muscle-specific gene expression. Taken together, these findings provide new insights into the molecular basis of vertebrate muscle fiber-type specification, and underscore Blimp1 as the central determinant of this process.


Correction: Photoelectron angular distributions as sensitive probes of surfactant layer structure at the liquid-vapor interface.
Correction for 'Photoelectron angular distributions as sensitive probes of surfactant layer structure at the liquid-vapor interface' by Rémi Dupuy et al., Phys. Chem. Chem. Phys., 2022, 24, 4796-4808, https://doi.org/10.1039/D1CP05621B.


Expression of the fragile (X) chromosome in an interspecific somatic cell hybrid.
Interspecific somatic cell hybrids were constructed between a Chinese hamster lung cell line deficient in hypoxanthine phosphoribosyltransferase and two lymphoblastoid cultures (GM 4025 and GM 3200) from unrelated males affected with the fragile (X) syndrome. Thirteen independent colonies survived selection in hypoxanthine-azaserine, while only one colony survived selection in hypoxanthine-aminopterin-thymidine. One hybrid formed from GM 4025 was found to contain a human X chromosome as the only detectable human chromosome in the majority of cells analyzed. Induction of fragile (X) expression in this hybrid at frequencies up to 20% was achieved by treatments with 5-fluoro-2'-deoxyuridine (5 X 10(-8) M or 1 X 10(-7) M) or methotrexate (5 X 10(-6) or 1 X 10(-5) for 12 h. Use of the somatic cell hybrid system may allow study of the fragile (X) from different patients on a homogeneous xenogeneic background and may provide a better system for characterization of the fragile (X) at the biochemical and molecular level.


Kinetics, Thermodynamics, and Mechanism of Cu(II) Ion Sorption by Biogenic Iron Precipitate: Using the Lens of Wastewater Treatment to Diagnose a Typical Biohydrometallurgical Problem.
The feasibility of improving typical biohydrometallurgical operation to minimize copper losses was investigated by the use of biogenic iron precipitate for the uptake of Cu(II) ions from aqueous solutions. The iron precipitate was obtained from mineral sulfide bioleaching and characterized using SEM/EDS, XRD, FTIR, BET, TGA, and pHpzc analyses. The results show that the precipitate is highly heterogeneous and that Cu(II) ion adsorption can be described by both Freundlich and Langmuir adsorption isotherms, with a maximum adsorption capacity of 7.54 mg/g at 30 °C and 150 mg/L. The sorption followed pseudo-second-order kinetics, while the major presence of -OH and -NH2 functional groups initiated a chemisorption mechanism through an ion-exchange pathway for the process. Ionic Cu(II) (radius (0.72 Å)) attached easily to the active sites of the precipitate than hydrated Cu(II) (radius (4.19 Å)). With an estimated activation energy of 23.57 kJ/mol, the obtained thermodynamic parameters of ΔS° (0.034-0.050 kJ/mol K), ΔG° (8.37-10.64 kJ/mol), and ΔH° (20.07-23.81 kJ/mol) indicated that the adsorption process was chemically favored, nonspontaneous, and endothermic, respectively. The 43% Cu(II) removal within 60 min equilibrium contact time at pH 5 was indicative of the reduced efficiency of copper extraction observed in a real-life biohydrometallurgical process due to sorption by the iron precipitate. The result of this study might provide an insight into the management of the biohydrometallurgical process to minimize copper losses. It may also help mitigate environmental pollution caused by the disposal of these biogenic iron precipitate residues.


Plasma N-terminal pro-brain natriuretic peptide levels are positively correlated with pulmonary arterial pressure in atrial septal defect patients.
N-terminal pro-brain natriuretic peptide (NT-proBNP), a pro-hormone secreted by the myocardium in response to various stimuli, was found to be correlated with several hemodynamic parameters in pulmonary hypertension associated with systemic sclerosis. We investigated plasma NT-proBNP levels and the relationships between NT-proBNP and several hemodynamic parameters in atrial septal defect (ASD) patients with or without pulmonary arterial hypertension (PAH). We found that plasma NT-proBNP level was significantly higher in PAH group compared with the control group (5495.4±388.4 pg/ml vs 4005.1±260.5 pg/ml, P<0.05). In a multiple regression model analysis, only mean pulmonary arterial pressure was an independent predictor of NT-proBNP (standardized coefficient=0.663, P=0.002). In the PAH group, only right atrial systolic pressure was found to be positively correlated with NT-proBNP, whereas other parameters were not found to be correlated with NT-proBNP. Our data suggests that NT-proBNP might also be a predictor of the severity of pulmonary hypertension in the ASD patients.


Semi-automated vs. manual 3D reconstruction of central mesenteric vascular models: the surgeon's verdict.
3D vascular anatomy roadmaps are currently being implemented for surgical planning and navigation. Quality of the reconstruction is critical. The aim of this article is to compare anatomical completeness of models produced by manual and semi-automatic segmentation. CT-datasets from patients included in an ongoing trial, underwent 3D vascular reconstruction applying two different segmentation methods. This produced manually-segmented models (MSMs) and semi-automatically segmented models (SAMs) which underwent a paired comparison. Datasets were delivered for reconstruction in 4 batches of 6, of which only batch 4 contained patients with abnormal anatomy. Model completeness was assessed quantitatively using alignment and distance error indexes and qualitatively with systematic inspection. MSMs were the gold standard. Assessed vessels were those of interest to the surgeon performing D3-right colectomy. 24 CT-datasets (13 females, age 44-77) were used in a paired comparative analysis of 48 3D-models. Quantitatively, SAMs showed structural improvement from Batch 1 to 3. Batch 4, with abnormal vessels, showed the highest error-index values. Qualitatively, 91.7% of SAMs did not contain all mesenteric branches relevant to the surgeon. In SAMs, 1 (12.5%) right colic artery-RCA scored as a complete vessel. 3 (37.5%) RCAs scored as incomplete and 4 (50%) RCAs were absent. 6 (25%) of 24 middle colic arteries-MCA scored as complete vessels. 11 (45.8%) scored as incomplete while 7 (29.2%) MCAs were absent. 13 (54.2%) of 24 ileocolic arteries-ICA were complete vessels. 11 (45.8%) scored as incomplete. None (0%) were absent. Additionally, it was observed that 10 (41.7%) of SAMs contained all their jejunal arteries, when compared to MSMs. Calibers of "complete" vessels were significantly higher than in "missing" vessels (MCA p < 0.001, RCA p = 0.016, ICA p < 0.001, JAs p < 0.001). Despite acceptable results from quantitative analysis, qualitative comparison indicates that semi-automatically generated 3D-models of the central mesenteric vasculature could cause considerable confusion at surgery.


Effect of physical activity and nutrition on bone mineral density in young Japanese women.
We explored factors that contributed to bone mineral density (BMD) in Japanese young women by quantifying the factors related to BMD. Between October 2003 and February 2004, we conducted a cross-sectional survey to study the status of nutritional intake and physical activity, and evaluated the various physical and serum parameters in relation to BMD. Subjects included 254 healthy female students who were 19-25 years old and were attending the Nursing School of Tokyo Women's Medical University, Japan. We measured the lumbar BMD (L2-L4) in these women. Multiple regression analysis was used to predict factors that contributed to current L2-L4 BMD. Our results showed that body mass index (BMI) (standardized regression coefficient = 0.45, P < 0.0001), past exercise habit (standardized regression coefficient = 0.15, P < 0.0059), and current total energy expenditure (standardized regression coefficient = 0.12, P < 0.03) were factors that significantly predicted the current L2-L4 BMD, with BMI as a key contributing factor. A BMI of 20.8 kg/m(2) allowed acquisition of young adult mean (YAM) irrespective of the total energy expenditure. In subjects with low BMI, L2-L4 BMD increased with higher current energy expenditure. A BMI of 20.8 kg/m(2) or greater and an energy expenditure of 32.9 METS-h/day or greater are required to acquire the YAM. We concluded that BMI and physical activity were factors that affected the BMD of Japanese young women.


Barriers to completion of health care proxies: an examination of ethnic differences.
Advance directives have not been uniformly used by different segments of the US population and studies have consistently shown a lower prevalence of advance directives among African Americans and Hispanics compared with non-Hispanic whites. To examine barriers to completion of health care proxies for different ethnic groups. One hundred ninety-seven subjects aged 65 years or older self-identified as African American (n = 65), Hispanic (n = 65), or non-Hispanic white (n = 67) attending a geriatrics and internal medicine outpatient clinic of a large New York City teaching hospital were administered a questionnaire. Questionnaires were developed to examine potential barriers to completion of health care proxies. Barriers were drawn from the literature and from focus groups. Significant predictors of proxy completion using logistic regression analysis included knowledge of health care proxies, availability of a health care agent, exposure to mechanical ventilation, age, and self-reported health status as fair to poor. Subjects who believed that a health care agent was irrelevant in the setting of involved family were significantly less likely to have completed a health care proxy. Although there were significant differences in the baseline completion rates of health care proxies for the 3 ethnic groups, ethnicity did not predict prior appointment of a health care agent in multivariate analysis. Differences in health care proxy completion rates across white, African American, and Hispanic elderly individuals in this New York City population seem to be related to potentially reversible barriers such as lack of knowledge and the perceived irrelevance of advance directives in the setting of involved family. Enhanced educational efforts of both health care personnel and patients could increase the rate of formal health care proxy appointment.


Inhibitors of sterol synthesis. Metabolism of 5 alpha-cholest-8(14)-en-3 beta-ol-15-one after intravenous administration to bile duct-cannulated rats.
The metabolism of 5 alpha-cholest-8(14)-en-3 beta-ol-15-one has been studied after intravenous administration to bile duct-cannulated rats. Very rapid and substantial conversion of the 15-ketosterol to polar biliary metabolites was observed in both male and female rats. For example, upon intravenous injection of [4-14C]5 alpha-cholest-8(14)-en-3 beta-ol-15-one to male bile duct-cannulated rats, approximately 86% of the administered 14C was recovered in bile in the first 38 h. Of the total amount of 14C recovered in bile in 38 h, approximately 50% was excreted in bile in the first 70 min and approximately 90% was excreted within 8 h after the injection of the 15-ketosterol. A substantial fraction of the polar biliary metabolites was shown to undergo enterohepatic circulation. Of the radioactivity derived from the labeled 15-ketosterol which was not recovered in bile or other excreta at 48 h after the intravenous administration of the 15-ketosterol, most (approximately 79%) was recovered in the form of cholesterol and cholesteryl esters of blood and the various tissues. The very substantial and rapid biliary excretion of polar metabolites of the 15-ketosterol (or of cholesterol derived from the 15-ketosterol), coupled with inhibition of the intestinal absorption of cholesterol by the 15-ketosterol, may contribute to the overall hypocholesterolemic action of the 15-ketosterol which has been observed in rodents and in nonhuman primates by providing a metabolic pathway(s) wherein a substantial fraction of the absorbed 15-ketosterol is rapidly removed from the body by biliary excretion in the form of polar metabolites.


The Biology of General Anesthesia from Paramecium to Primate.
General anesthesia serves a critically important function in the clinical care of human patients. However, the anesthetized state has foundational implications for biology because anesthetic drugs are effective in organisms ranging from paramecia, to plants, to primates. Although unconsciousness is typically considered the cardinal feature of general anesthesia, this endpoint is only strictly applicable to a select subset of organisms that are susceptible to being anesthetized. We review the behavioral endpoints of general anesthetics across species and propose the isolation of an organism from its environment - both in terms of the afferent arm of sensation and the efferent arm of action - as a generalizable definition. We also consider the various targets and putative mechanisms of general anesthetics across biology and identify key substrates that are conserved, including cytoskeletal elements, ion channels, mitochondria, and functionally coupled electrical or neural activity. We conclude with a unifying framework related to network function and suggest that general anesthetics - from single cells to complex brains - create inefficiency and enhance modularity, leading to the dissociation of functions both within an organism and between the organism and its surroundings. Collectively, we demonstrate that general anesthesia is not restricted to the domain of modern medicine but has broad biological relevance with wide-ranging implications for a diverse array of species.


Inactivation of Bacillus spores in reconstituted skim milk by combined high pressure and heat treatment.
To determine the resistance of a variety of Bacillus species spores to a combined high pressure and heat treatment; and to determine the affect of varying sporulation and treatment conditions on the level of inactivation achieved. Spores from eight Bacillus species (40 isolates) were high pressure-heat treated at 600 MPa, 1 min, initial temperature 72 degrees C. The level of inactivation was broad (no inactivation to 6 log10 spores ml(-1) reduction) and it varied within species. Different sporulation agar, high pressure equipment and pressure-transmitting fluid significantly affected the response of some isolates. Varying the initial treatment temperature (75, 85 or 95 degrees C) shifted the relative order of isolate high pressure-heat resistance. The response of Bacillus spores to combined high pressure-heat treatment is variable and can be attributed to both intrinsic and extrinsic factors. The combined process resulted in a high level of spore inactivation for several Bacillus species and is a potential alternative treatment to traditional heat-only processes. Sporulation conditions, processing conditions and treatment temperature all affect the response of Bacillus spores to the combined treatment of high pressure and heat. High levels of spore inactivation can be achieved but the response is variable both within and between species.


Reading the glyco-code: New approaches to studying protein-carbohydrate interactions.
The surface of all living cells is decorated with carbohydrate molecules. Hundreds of functional proteins bind to these glycosylated ligands; such binding events subsequently modulate many aspects of protein and cell function. Identifying ligands for glycan-binding proteins (GBPs) is a defining challenge of glycoscience research. Here, we review recent advances that are allowing protein-carbohydrate interactions to be dissected with an unprecedented level of precision. We specifically highlight how cell-based glycan arrays and glyco-genomic profiling are being used to define the structural determinants of glycan-protein interactions in living cells. Going forward, these methods create exciting new opportunities for the study of glycans in physiology and disease.


Women's experience of menopause in rural communities in Orlu, Eastern Nigeria.
To document women's experience of menopause in rural communities in Orlu, Eastern Nigeria, and to compare this with urban centers in Nigeria and worldwide. A total of 349 women who had not menstruated for at least 12 months were included in a questionnaire-based study. The questionnaire included questions on sociodemographic characteristics, age at menarche and menopause, attitude to menopause, and knowledge and use of hormone replacement therapy. Questionnaires were administered in 13 communities by medical students during the Annual Convention of Christian Women in August 2009. The mean age at menopause of the women was 47 ± 4.2 years. Menopausal symptoms were prevalent, and bone and joint pain were the most prevalent symptoms. Knowledge and use of hormone replacement therapy were poor. Most of the women considered the menopause to be beneficial. Age at menopause and associated postmenopausal symptoms were similar to those seen in urban centers in Nigeria, but age at menopause was lower than in studies of white women. Despite the apparent positive attitude of Nigerian women to menopause, a program of information and education, and use of hormone replacement therapy will improve quality of life for women in Eastern Nigeria.


Serum and aqueous humour concentration of tranexamic acid after peroral administration.
After a single oral dose of 25 mg/kg tranexamic acid in 37 patients with cataract as the sole eye disease, the concentration of the drug was measured in both the serum and the aqueous at various intervals following intake. The serum concentration was highest after three h (average 15.44 mg/l), but a trace of tranexamic acid (0.7 mg/l) could still be found after 19 h. In the aqueous, the concentration was likewise highest after three h (average 1.62 mg/l), but, following this, the fall in tranexamic acid concentration was very gradual and after 19 h was found to be 1.3 mg/l. In two patients who had received 25 mg/kg three times daily for three days, an aqueous concentration of 2.3 mg/l was found eight h after the final intake.


[Calculation of the probability of paternity and of the chance of paternity exclusion for the HLA system using only the typing results from the child and the putative father (author's transl)].
A method for the calculation of the probability of paternity for the HLA system using only the typing results from the child and the putative father, without taking into account the data of the mother, is presented. Furthermore, the usability of the formulas by Mayr and Pausch (Z. Immun.-Forsch. 150, 447 (1975)) for the computation of the chance of paternity exclusion in such cases is demonstrated.


Growth and stability of rocksalt Zn1-xMgxO epilayers and ZnO/MgO superlattice on MgO (100) substrate by molecular beam epitaxy.
Zn1-xMgxO films with x = 0.04-0.50 grown on MgO (100) substrates by molecular beam epitaxy retain the rocksalt (rs) crystal structure and grow epitaxially for x ≥ 0.17. In addition, the rs-ZnO epilayer is observed to be stable up to a thickness of 5 nm and also in a ZnO/MgO superlattice sample. However, a portion of the superlattice has transformed to wurtzite (wz)-structure islands in a self-accommodated manner during growth. The transformation is a combination of a Bain distortion, an in-plane rotation of 14.5°, and a Peierls distortion, resulting in an orientation relationship of (100)rs//(101̄0)wz and 〈011〉rs ∼//〈1̄21̄3〉wz. In such a manner, the volume expansion is only necessary along the growth direction and the in-plane strains can be minimized. A negative pressure generated during the transformation of ZnO stabilizes the MgO into a wurtzite structure.


Guilu Erxian Glue () Inhibits Chemotherapy-Induced Bone Marrow Hematopoietic Stem Cell Senescence in Mice May via p16INK4a-Rb Signaling Pathway.
To evaluate the effect of Guilu Erxian Glue (, GEG) on cyclophosphamide (CTX)-induced bone marrow hematopoietic stem cells (HSCs) senescence in mice and explore the underlying mechanism. The H22 liver cancer ascites lump model was established in male Kunming mice by injecting intraperitoneally (i.p.) with 5 × 106/mL H22 cells per mouse. Fifty tumor-bearing mice were divided into the control, model, pifithrin-α, GEG, and GEG+pifithrin-α groups using a random number table, 10 mice in each group. CTX (100 mg/kg i.p.) was administrated to mice from day 1 to day 3 (d1-d3) continuously except for the control group. The mice in the pifithrin-α, GEG and GEG+pifithrin-α groups were treated with pifithrin-α (2.2 mg/(kg·d) i.p.) for 6 consecutive days (d4-d9), GEG (9.5 g/(kg·d) i.p.) for 9 consecutive days (d1-d9), and GEG plus pifithrin-α, respectively. HSCs were collected after 9-d drug treatment. The anti-aging effect of GEG was studied by cell viability, cell cycle, and β -galactosidase (β -gal) assays. The mRNA and protein expressions of cyclin-dependent kinase 2 (CDK2), CDK4, inhibitor of cyclin-dependent kinase 4a encoding the tumor suppressor protein p16 (p16INK4a), p21Cip1/Waf1, p53, and phosphorylated retinoblastoma (pRb) were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and semi-quantitative Western blot, respectively. Compared with the model group, GEG increased cell viability as well as proliferation (P<0.05 or P<0.01) and reduced β -gal expression. Furthermore, GEG significantly decreased the expressions of p16INK4a, p53 and p21Cip1/Waf1 proteins, and increased the expressions of CDK2, CDK4 and pRb proteins compared with the model group (P<0.05 or P<0.01). GEG can alleviate CTX-induced HSCs senescence in mice, and the p16INK4a-Rb signaling pathway might be the underlying mechanism.


CLDN-1 promoted the epithelial to migration and mesenchymal transition (EMT) in human bronchial epithelial cells via Notch pathway.
Claudin-1 (CLDN-1) is one of main tight junction components that play an important role in epithelial-mesenchymal transition (EMT). However, the effects of CLDN-1 on the migration and EMT induced by TGF-β1 in primary normal human bronchial epithelial (NHBE) and BEAS-2B cells have not been clear. The expression of CLDN-1 was quantified by Western blotting in NHBE and BEAS-2B cells. Cell migration and invasion were detected using transwell assays. The expression level of E-cadherin, N-cadherin, α-SMA, and Vimentin was evaluated by quantitative real-time PCR and Western blotting. Here we showed that the protein expression of CLDN-1 was increased exposed to TGF-β1 in a dose- and time-dependent manner. Knockdown of CLDN-1 using small interfering CLDN-1 RNA (siCLDN-1) prevented the migration and invasion in NHBE and BEAS-2B cells. Moreover, depletion of CLDN-1 promoted the E-cadherin expression and decreased the mRNA and protein levels of N-cadherin, α-SMA, and Vimentin induced by TGF-β1. Furthermore, CLDN-1 silencing resulted in the reduction of the Notch intracellular domain (NICD) and hairy enhancer of split-1 (Hes-1) in mRNA and protein level. Jagged-1, an activator of Notch signaling pathway, abrogated the protective function of siCLDN-1 in migration and EMT. In conclusion, CLDN-1 promoted the migration and EMT through the Notch signaling pathway.


Compact conformal tattoo-polymer antenna for on-body wireless power transfer.
This paper presents a 35.0 × 35.0 × 2.7 mm3 compact, low-profile, and lightweight wearable antenna for on-body wireless power transfer. The proposed antenna can be easily printed on a piece of flexible tattoo paper and transformed onto a PDMS substrate, making the entire antenna structure conform to the human body for achieving a better user experience. Here, a layer of frequency selective surface (FSS) is inserted in between the antenna and human tissue, which has successfully reduced the loading effects of the tissue, with 13.8 dB improvement on the antenna gain. Also, the operating frequency of the rectenna is not affected much by deformation. To maximize the RF-DC conversion efficiency, a matching loop, a matching stub, and two coupled lines are integrated with the antenna for tuning the rectenna so that a wide bandwidth (~ 24%) can be achieved without the use of any external matching networks. Measurement results show that the proposed rectenna can achieve a maximum conversion efficiency of 59.0% with an input power of 5.75 μW/cm2 and can even exceed 40% for a low input power of 1.0 μW/cm2 with a 20 kΩ resistive load, while many other reported rectennas can only achieve a high PCE at a high power density level, which is not always practical for a wearable antenna.


The posterior pituitary of the garden dormouse (Eliomys quercinus l.). Evidence of two types of neurosecretory axons on the basis of ultrastructural characteristics.
Ultrastructural examination of the posterior pituitary of the garden dormouse (Eliomys quercinus L) was carried out at different times in the annual cycle of this hibernating rodent. Obvious differences between experimental groups have not been observed, and the results presented here must be considered as general features of the garden dormouse posterior pituitary. Neurosecretory axons and endings can be divided into two types, according to different aspects of neurosecretory granules (NSG) and microvesicles (MV). One type contains spherical NSG with homogeneous cores and round MV. In the other type, NSG have various, often elongated, shapes. Their content shows two types of crystalline structures and most of the MV have flattened aspects. As it is very unlikely that this duality in NSG is a result of an artefact of fixation, three hypotheses are presented as explanation. The duality of NSG might be related either to their hormonal content (oxytocin or vasopressin) or to their degree of maturation. Moreover, both explanations may be valid. In the species studied, pituicytes often contain concentric lamellar structures of the endoplasmic reticulum (whorls), the significance of which remains obscure.


A comparative study of mouse nasal septal and turbinal epithelium for in vitro cell cultures.
The small number of epithelial cells that can be isolated from mouse respiratory tract has limited its application to study of the airway. Our purpose in this work was to compare the potential of mouse nasal septal and turbinal epithelium for serving as a cell source for in vitro cell cultures. The distribution of ciliated epithelial cells in the mouse nasal cavity, and the surface area of the respiratory epithelium in the mouse nasal septum and turbinate were determined by β-tubulin IV immunohistochemistry. Ciliated epithelial cell density of native mouse nasal septum and turbinate, and the morphology of cultured mouse nasal septal and turbinal epithelial cells were examined by scanning electron microscopy (SEM) and β-tubulin IV and zona occludens (ZO)-1 dual-label fluorescent cytochemistry, respectively. Cilia reactivity to exogenous stimuli (adenosine triphosphate or benzalkonium chloride) in cultured nasal septal and turbinal epithelial cells was studied using high-speed digital microscopy. The ciliated epithelial cell density of native nasal epithelium, the morphology and cell yield of in vitro cultured epithelial cells, and cilia reactivity to exogenous stimuli of mouse turbinate are comparable to that of the nasal septum. However, the respiratory epithelium surface area of mouse turbinate is significantly larger than that of the nasal septum. Mouse turbinate may also serve as an ideal source of in vitro epithelial cell cultures. Both nasal septum and turbinate should be harvested to optimize animal use in the establishment of primary murine respiratory epithelial cultures.


STAT3 regulates miR93-mediated apoptosis through inhibiting DAPK1 in renal cell carcinoma.
Signal transducer and activator of transcription 3 (STAT3) is an essential member of the STAT family. STAT3 regulates diverse genes that mediate inflammatory reactions, cell survival, proliferation, and angiogenesis, and it is aberrantly upregulated and activated in various types of malignancies. Furthermore, STAT3 signalling is involved in multiple feedback loops and pathways. In this study, we demonstrate that miR-93-3p plays an oncogenic role in renal cell carcinoma (RCC) by enhancing RCC cell proliferation and suppressing apoptosis. In addition, STAT3 can regulate the transcription of miR-93 by directly binding its promoter region. miR-93 can inhibit death-associated protein kinase 1 (DAPK1) at the protein level. Moreover, STAT3 can block DAPK1 expression at the RNA level. Importantly, we verified that DAPK1 overexpression in turn suppresses the entry of activated STAT3 into the cell nucleus. Thus, this study reveals a potential continuously activated signalling transduction pathway, STAT3-miR93-DAPK1, and may provide a novel clinical therapeutic approach for RCC.


Maternal α-linolenic acid availability during gestation and lactation alters the postnatal hippocampal development in the mouse offspring.
The availability of ω-3 polyunsaturated fatty acids is essential for perinatal brain development. While the roles of docosahexaenoic acid (the most abundant ω-3 species) were extensively described, less is known about the role of α-linolenic acid (ALA), which is the initial molecular species undergoing elongation and desaturation within the ω-3 pathways. This study describes the association between maternal ALA availability during gestation and lactation, and alterations in hippocampal development (dentate gyrus) in the mouse male offspring, at the end of lactation (postnatal day 19, P19). Postnatal ALA supplementation increased cell proliferation (36% more proliferating cells compared to a control group) and early neuronal differentiation, while postnatal ALA deficiency increased cellular apoptosis within the dentate gyrus of suckling pups (61% more apoptotic cells compared to a control group). However, maternal ALA deficiency during gestation prevented the increased neurogenesis induced by postnatal supplementation. Fatty acid analysis revealed that ALA supplementation increased the concentration of the ω-3 species in the maternal liver and serum, but not in the brain of the offspring, excepting for ALA itself. Interestingly, ALA supplementation also increased the concentration of dihomo γ-linolenic acid (a ω-6 species) in the P19 brains, but not in maternal livers or serum. In conclusion, postnatal ALA supplementation enhances neurogenesis in the dentate gyrus of the offspring at postnatal day 19, but its beneficial effects are offset by maternal ALA deficiency during gestation. These results suggest that ALA is required in both fetal and postnatal stages of brain development.


A review of the composition, characteristics, and effectiveness of barrier mesh prostheses utilized for laparoscopic ventral hernia repair.
The objective of this review was to provide an overview of the components that comprise each of the eight barrier mesh prostheses commonly utilized for LVHR and to review the current literature related to the characteristics and effectiveness of these materials to guide the general surgeon in selecting the most appropriate material for LVHR. Composite prostheses with permanent barriers (Bard™ Composix™ E/X, Bard™ Composix™ L/P, and DUALMESH(®) Biomaterial) were compared to composite prostheses with absorbable barriers (C-QUR™ Mesh, PROCEED™ Surgical Mesh, Bard™ Sepramesh™ IP Composite, Parietex™ Composite, and PHYSIOMESH™) using scanning electron microscopy and a review of the current preclinical and clinical literature. Clinical studies and preclinical animal models have attempted to determine the adhesion characteristics and effectiveness of barrier mesh prostheses available for ventral hernia repair applications. However, it is difficult to make any definitive statements about the adhesion characteristics and effectiveness of these materials because all meshes were not included in all studies and likewise not compared under identical conditions. Overall, Parietex™ Composite and DUALMESH(®) Biomaterial were cited most frequently for improvement of adhesion characteristics, followed closely by Bard™ Sepramesh™ IP Composite and C-QUR™ Mesh. Bard™ Composix™, PROCEED™ Surgical Mesh, and uncoated polypropylene were cited most frequently as having the most tenacious and extensive adhesions. Differences observed between the various barrier prostheses are likely attributable to the chemical composition of the barrier or the conditions required for resorption and metabolism of the barrier components. It is likely that the components of these barriers incite a wide range of inflammatory responses resulting in the range of adhesion coverage and tenacity observed in the preclinical and clinical studies reviewed. Clinical trials are needed to more appropriately define the clinical effectiveness of these barriers.


Synthesis and triplex forming ability of conformationally locked oligonucleotides containing unnatural nucleobases: efficient recognition of a C.G interruption.
In order to develop a novel nucleoside analogue which recognizes C.G interruption in homopurine.homopyrimidine DNA, we designed and synthesized a conformationally locked nucleoside analogue, 1-(2-O,4-C-methylene-beta-D-ribofuranosyl)pyridin-2-one (4), and introduced it into a triplex-forming oligonucleotide (TFO). On melting temperature (Tm) measurements, the unprecedented C.G base recognition ability of 4 was observed.


Omission of Completion Lymph Node Dissection in Sentinel Node Biopsy Positive Head and Neck Cutaneous Melanoma Patients.
Recent studies evaluating patients with a positive sentinel lymph node biopsy (SLNB+) show no melanoma-specific survival difference between patients undergoing lymph node basin surveillance and completion lymph node dissection (CLND). This has been broadly applied, despite underrepresentation of head and neck (HN) cutaneous melanoma patients. We evaluated whether this was upheld in the HN melanoma cohort. Patients with HN melanoma with a SLNB+ were selected from the National Cancer Database (NCDB) from 2012 to 2019. Overall survival (OS) of patients who underwent SLNB only versus SLNB + CLND were compared. Subgroup analyses were performed based on pathologic N (pN) and receipt of immunotherapy. Adjusted hazard ratio (aHR) and 95% confidence interval (CI) were calculated. Analysis of 634 patients with multivariable Cox regression showed no difference in OS in SLNB only versus SLNB + CLND cohorts (hazard ratio [HR] 1.13; 95% confidence interval [CI] 0.71-1.81; p = 0.610). Charlson-Deyo score (CDS) 1 versus 0 (HR 1.70; 95% CI 1.10-2.63; p = 0.016), pN2+ versus pN1 (HR 1.74; 95% CI 1.23-2.45; p = 0.002), and lymphovascular invasion (LVI) versus no (HR 2.07; 95% CI 1.34-3.19; p = 0.001) were associated with worse prognosis. Subgroup analysis by pN showed no OS benefit for CLND in either pN1 (HR 1.04; 95% CI 0.51-2.10; p = 0.922) or pN2+ (HR 1.31; 95% CI 0.67-2.57; p = 0.427) patients or in patients who received immunotherapy (HR 1.32; 95% CI 0.54-3.22; p = 0.549). This study of SLNB + HN melanoma patients showed no OS difference in SLNB only versus SLNB + CLND. Further studies need to be performed to better define the role of CLND.


Targeting arginase-1 exerts antitumor effects in multiple myeloma and mitigates bortezomib-induced cardiotoxicity.
Multiple myeloma (MM) remains an incurable malignancy of plasma cells despite constantly evolving therapeutic approaches including various types of immunotherapy. Increased arginase activity has been associated with potent suppression of T-cell immune responses in different types of cancer. Here, we investigated the role of arginase 1 (ARG1) in Vκ*MYC model of MM in mice. ARG1 expression in myeloid cells correlated with tumor progression and was accompanied by a systemic drop in ʟ-arginine levels. In MM-bearing mice antigen-induced proliferation of adoptively transferred T-cells was strongly suppressed and T-cell proliferation was restored by pharmacological arginase inhibition. Progression of Vκ*MYC tumors was significantly delayed in mice with myeloid-specific ARG1 deletion. Arginase inhibition effectively inhibited tumor progression although it failed to augment anti-myeloma effects of bortezomib. However, arginase inhibitor completely prevented development of bortezomib-induced cardiotoxicity in mice. Altogether, these findings indicate that arginase inhibitors could be further tested as a complementary strategy in multiple myeloma to mitigate adverse cardiac events without compromising antitumor efficacy of proteasome inhibitors.


Participation of PI3-kinase/Akt signalling in insulin stimulation of p34cdc2 activation in zebrafish oocyte: phosphodiesterase 3 as a potential downstream target.
Exposure of fully grown oocytes to growth factors (insulin/IGFs) initiates various signalling cascades that culminate to final stages of oocyte maturation. Regulation of signalling pathways during growth factor-induced meiosis resumption in fish is not well characterized. Here we studied the participation of PI3K/Akt signalling pathway during recombinant human insulin (rh-insulin)-induced meiotic maturation in zebrafish (Danio rerio) oocytes. Priming of defolliculated oocytes in vitro with rh-insulin promotes germinal vesicle breakdown (GVBD) in a dose- and time-dependent manner, an effect sensitive to translation but not transcription inhibition. More than 80% of the oocytes underwent GVBD due to 0.8IU/ml rh-insulin within 10h of incubation and the kinetics of p34cdc2 kinase activation corresponded well with GVBD data. PI3K inhibitors, wortmannin and LY294002 blocked insulin, but not 17α, 20β-DHP-induced GVBD. Immunoblot analyses of oocyte extract revealed that phospho-PI3K (p85α) was up regulated within 30-60 min of insulin stimulation followed by phospho-Akt (Ser473) at 60-120 min. Though PI3K/Akt phosphorylation was largely unaffected, pre-incubation with phosphodiesterase (PDE) inhibitors, IBMX and cilostamide, but not rolipram completely blocked rh-insulin-induced p34cdc2 activation and GVBD. These results suggest that PDE3 may be one potential downstream target to PI3K/Akt signalling necessary for rh-insulin-induced GVBD in zebrafish.


Intra-aortic balloon pumping: effects on left ventricular diastolic function.
The intra-aortic balloon pump is the most widely used form of temporary cardiac assist and often utilised in patients before and after cardiac surgery. Several effects of balloon counter-pulsation have been reported previously, but its effect on left ventricular diastolic function has not been thoroughly investigated. The aim of this study is to examine the effect of the intra-aortic balloon pump on left ventricular wall motion and transmitral flow. We studied 20 patients in the intensive care unit, less than 36 h following cardiac surgery. We recorded left anterior descending coronary artery and transmitral E-wave flow velocities using transesophageal echocardiography pulsed Doppler. We also recorded left ventricular long axis free-wall movement using M-mode. The intra-aortic balloon pump was set to full augmentation and recordings were made at pumping cycles 1:1, 1:2, 1:3, and when the pump was on stand-by, leaving a minimum of 5 min between the pumping modes to allow the return to control conditions. In order to eliminate time effects, the sequence of recording was varied between patients using a 4 by 4 Latin-square. The peak diastolic left anterior descending coronary artery and transmitral E-wave flow velocities, and left ventricular free-wall early diastolic lengthening velocity increased significantly with intra-aortic balloon pumping cycles 1:1, 1:2 and 1:3 compared to their value with the pump on stand-by, all P < 0.001. The increase in peak transmitral E-wave flow velocity correlated with the increase in peak left anterior descending coronary artery diastolic flow velocity (r = 0.74, P = 0.02), and with the increase in left ventricular free-wall early diastolic lengthening velocity (r = 0.80, P < 0.001). Using the intra-aortic balloon pump post-cardiac surgery significantly increases peak diastolic left anterior descending coronary artery flow velocities and left ventricular free-wall early diastolic lengthening velocity, whose increase explains the increase in peak transmitral E-wave velocity. Although coronary flow is epicardial and mitral flow is intracardial, their close relationship suggests an improvement in left ventricular diastolic function with intra-aortic balloon pump.