Nanocrystals of magnesium and fluoride substituted hydroxyapatite.
Hydroxyapatite nanocrystals synthetized in the presence of different concentrations of magnesium and fluoride ions in solutions--1, 5 and 10 at.% have been submitted to a structural and chemical characterization. The syntheses were carried out in the presence of low molecular weight polyacrylic acid, which has been verified to inhibit hydroxyapatite crystallization. The polyelectrolyte is adsorbed into the crystals during the synthesis and provokes a reduction of the mean crystal sizes. The reduction is greater along the direction orthogonal to the c-axis, suggesting a preferential adsorption of the polyelectrolyte on the crystalline faces parallel to the c-axis. Both magnesium and fluoride can be incorporated into the hydroxyapatite structure. On the basis of the values of the lattice constants and of the magnesium relative content of the solid phase, it can be suggested that probably just a part of magnesium is substituted for calcium, the remainder being adsorbed on the crystal surface. However, magnesium destabilizes the apatitic structure favouring its thermal conversion into beta-tricalcium phosphate, and displays an inhibiting effect on the crystallization of hydroxyapatite. This last effect is enhanced by the simultaneous presence of polyacrylic acid. Fluoride substitution for hydroxyl ions into hydroxyapatite structure induces a slight increase of the crystal sizes along the c-axis direction. The data indicate that the experimental approach can be successfully used to prepare nanoapatite with crystallinity, crystal dimensions, composition, structure and stability very close to those characteristics of biological apatites.


Single-cell analysis of autophagy activity in normal and de novo transformed human mammary cells.
Assessment of autophagy activity has historically been limited to investigations of fixed tissue or bulk cell populations. To address questions of heterogeneity and relate measurements to functional properties of viable cells isolated from primary tissue, we created a lentiviral (RFP-GFP-MAP1LC3B) vector that allows the autophagosome and autolysosome content of transduced cells to be monitored at the single-cell level. Use of this strategy to analyze purified subsets of normal human mammary cells showed that both the luminal progenitor-containing (LP) subset and the basal cells (BCs) display highly variable but overall similar autophagic flux activity despite differences suggested by measurements of the proteins responsible (i.e., LC3B, ATG7 and BECLIN1) in bulk lysates. Autophagosome content was also highly variable in the clonogenic cells within both the LPs and BCs, but the proliferative response of the BCs was more sensitive to autophagy inhibition. In addition, use of this vector showed cells with the lowest autophagosome content elicited the fastest tumor growth in 2 different models of human mammary tumorigenesis. These results illustrate the utility of this vector to define differences in the autophagy properties of individual cells in primary tissue and couple these with their responses to proliferative and oncogenic stimuli.


Novel NaPi-IIc mutations causing HHRH and idiopathic hypercalciuria in several unrelated families: long-term follow-up in one kindred.
Homozygous and compound heterozygous mutations in SLC34A3, the gene encoding the sodium-dependent co-transporter NaPi-IIc, cause hereditary hypophosphatemic rickets with hypercalciuria (HHRH), a disorder characterized by renal phosphate-wasting resulting in hypophosphatemia, elevated 1,25(OH)(2) vitamin D levels, hypercalciuria, rickets/osteomalacia, and frequently kidney stones or nephrocalcinosis. Similar albeit less severe biochemical changes are also observed in heterozygous carriers, which are furthermore indistinguishable from those encountered in idiopathic hypercalciuria (IH). We now searched for SLC34A3 mutations (exons and introns) in two previously not reported HHRH kindreds, which resulted in the identification of three novel mutations. The affected members of kindred A were compound heterozygous for two different mutations, c.1046_47del and the intronic mutation c.560+23_561-42del, while the index case in kindred B was homozygous for the nonsense SLC34A3 mutation c.1764C>G (p.Y588X). The patient in kindred C was diagnosed with IH because of bilateral medullary nephrocalcinosis, suppressed PTH levels, and hypercalciuria; she was found to have a novel heterozygous c.1571_1880del mutation. The HHRH patients in kindred A were treated for up to 7years with oral phosphate, which led to reversal of hypophosphatemia, hypercalciuria, and prevention or healing of the mild bone abnormalities. PTH levels were normal throughout the observation period, while 1,25(OH)(2) vitamin D levels remained elevated and may thus be helpful for assessing treatment efficacy and patient compliance in HHRH.


Fine needle aspiration of thyroid: a cyto-histopathological correlation in Ramathibodi Hospital.
To evaluate the accuracy of fine needle aspiration cytology (FNAC) of thyroid lesions in Ramathibodi patients. A retrospective review is performed on 469 cases of thyroid surgical pathology specimens with previous FNAC reports; during January 2005 to January 2008. All histopathology reports are compared to the latest previous cytopathology results which are categorized as unsatisfactory, benign, inconclusive, borderline and malignancy The cytopathology results showed 74 cases (15.7%) of unsatisfactory specimens, 243 cases (51.8%) of benign lesions, 33 cases (7%) of inconclusive lesions, 15 cases (3.2%) of borderline lesions and 104 cases (22%) of malignant lesions. The histopathology results for benign lesions reveal multinodular and nodular goiter of 236 cases and other diagnosis of 54 cases. The histopathology results for malignant lesions were papillary carcinoma of 147 cases, follicular carcinoma of 24 cases and other malignancy of 8 cases. Among 243 cases of benign from FNAC; 23 cases turn out to be malignancy while 104 cases of malignancy from FNAC show no false positive. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of FNAC are 81.9%, 100%, 100%, 90.4% and 93.4% respectively. FNAC is a minimally invasive, highly accurate and cost-effective procedure. In our setting the FNAC enable the clinician to "rule-in" malignant lesions with confidence.


Upregulation of Thrombospondin 1 Expression in Synovial Tissues and Plasma of Rheumatoid Arthritis: Role of Transforming Growth Factor-β1 toward Fibroblast-like Synovial Cells.
To investigate the role of thrombospondin 1 (TSP-1) in RA. Expression of TSP-1 in synovial tissues was determined by immunohistochemistry. Expression of TSP-1 in rheumatoid fibroblast-like synovial cells (FLS) was investigated by quantitative real-time PCR and ELISA. Correlations among the plasma TSP-1 and other variables in patients with RA were examined. Expression of TSP-1 was increased in rheumatoid synovial tissues. Transforming growth factor-β1 (TGF-β1) clearly increased TSP-1 expression in FLS on both mRNA and protein levels. Changes in plasma TSP-1 were associated with those in 28-joint Disease Activity Score-erythrocyte sedimentation rate and plasma TGF-β1. TSP-1 might be critically involved in the disease process of RA through the TGF-β1/TSP-1 axis.


Quantitative analysis of O6-methylguanine DNA methyltransferase (MGMT) promoter methylation in patients with low-grade gliomas.
Methylation of the MGMT promoter is supposed to be a predictive and prognostic factor in glioblastoma. Whether MGMT promoter methylation correlates with tumor response to temozolomide in low-grade gliomas is less clear. Therefore, we analyzed MGMT promoter methylation by a quantitative methylation-specific PCR in 22 patients with histologically verified low-grade gliomas (WHO grade II) who were treated with temozolomide (TMZ) for tumor progression. Objective tumor response, toxicity, and LOH of microsatellite markers on chromosomes 1p and 19q were analyzed. Histological classification revealed ten oligodendrogliomas, seven oligoastrocytomas, and five astrocytomas. All patients were treated with TMZ 200 mg/m2 on days 1-5 in a 4 week cycle. The median progression-free survival was 32 months. Combined LOH 1p and 19q was found in 14 patients; one patient had LOH 1p alone and one patient LOH 19q alone. The LOH status could not be determined in two patients and was normal in the remaining four. LOH 1p and/or 19q correlated with longer time to progression but not with radiological response to TMZ. MGMT promoter methylation was detectable in 20 patients by conventional PCR and quantitative analysis revealed the methylation status was between 12 and 100%. The volumetric response to chemotherapy analyzed by MRI and time to progression correlated with the level of MGMT promoter methylation. Therefore, our retrospective case series suggests that quantitative methylation-specific PCR of the MGMT promoter predicts radiological response to chemotherapy with TMZ in WHO grade II gliomas.


CCR5 N-terminus peptides enhance X4 HIV-1 infection by CXCR4 up-regulation.
The HIV-1 envelope glycoprotein gp120 interacts consecutively with CD4 and CCR5 to mediate the entry of R5-HIV-1 strains into target cells. The N-terminus of CCR5, which contains several sulfated tyrosines, plays a critical role in gp120-CCR5 binding and, consequently, in viral entry. Here, we demonstrate that a tyrosine sulfated peptide, reproducing the entire N-terminal extracellular region of CCR5, its unsulfated analogue, and a point-mutated peptide are unable to inhibit R5-HIV-1 mediated infection, competing with the entire CCR5 in the formation of gp120-CD4-CCR5 complex. Surprisingly, these peptides show the capability of enhancing HIV-1 infection caused by X4 strains through the up-regulation of both CD4 and CXCR4 receptors.


Endothelial cell activation by interleukin-1 and extracellular matrix laminin-10 occurs via the YAP signalling pathway.
Laminin-10 (LM-10) is a key regulator of blood-brain barrier (BBB) repair after hypoxia and inflammation. Here we investigated the signalling mechanisms regulated by LM-10 in human brain endothelial cell line hCMEC/D3 in response to interleukin(IL)-1beta(β) in vitro. LM-10 promoted endothelial proliferation and repair of an endothelial monolayer after scratch injury, and upregulated IL-1β-induced ICAM-1 and VCAM-1 expression. IL-1β and LM-10 regulated YAP signalling pathway in endothelial cells leading to differential expression of YAP target genes, ctgf and serpine-1, providing evidence that the YAP signalling pathway could be a new therapeutic target for the treatment of BBB dysfunction in CNS diseases.


Are awareness questionnaires valid? Investigating the use of posttest questionnaires for assessing awareness in implicit memory tests.
Two experiments--one employing a perceptual implicit memory test and the other a conceptual implicit memory test--investigated the validity of posttest questionnaires for determining the incidence of awareness in implicit memory tests. In both experiments, a condition in which none of the studied words could be used as test responses (i.e., the none-studied condition) was compared with a standard implicit test condition. Results showed that reports of awareness on the posttest questionnaire were much less frequent in the none-studied condition than in the standard condition. This was especially true after deep processing at study. In both experiments, 83% of the participants in the none-studied condition stated they were unaware even though there were strong demands for claiming awareness. Although there was a small bias in the questionnaire (i.e., 17% of the participants in the none-studied condition stated they were aware), overall, there was strong support for the validity of awareness questionnaires.


Parathion effects on protein synthesis in the seminiferous tubules of mice.
Agropesticides impart high risks of exposure for animals including humans. This work analysed the effect of parathion upon protein synthesis in seminiferous tubules cultured in vitro from adult male CF1 mice. They were incubated with parathion solutions (0.8, 0.4, 0.04, 0.004, 0.0004 mM) for 5 h. One hour after ending incubation tritiated leucine was added to each sample. In addition, groups treated with 0.8 mM parathion were incubated for 5 h, and thereafter transferred to a parathion-free medium; tritiated leucine incorporation was measured 1, 2, or 3 h afterwards. Results of protein synthesis denote an inhibitory dose-dependent effect of parathion. This is reversed after pesticide-free incubation. Testicular protein synthesis inhibition induced by parathion may compromise male fertility in view of the fact that, in spermatogenesis, proliferation (mitosis, meiosis) and cell differentiation occurs.


[Gastrin].
Gastrin antisera were raised by immunization of rabbits or guinea pigs with synthetic human gastrin I conjugated to bovine serum albumin by carbodiimide. Radioiodination of SHG: 2-17 was performed by the chloramine T-method and by an enzymatic procedure. AE-cellulose was used to get a monoiodinated tracer hormone. Antibody reactions with the different forms of gastrin and with CCK-PZ was characterized. Precision (VK = 6-8%) and reproducibility (VK less than 15%) of the gastrin values were comparable to the insulin assay. Gastrin stimulates the parietal cell and has trophic effects on gastric mucosa. Hypergastrinaemia in combination with hypersecretion exhibits clinical significance in patients suffering from Zollinger-Ellison-syndrome or excluded antrum-syndrome which are due to autonomous gastrin release. Some findings suggest a pathogenetic role of gastrin in duodenal ulcer disease. Those disease in which gastrin determinations are of clinical value are discussed.


Fast Food Pattern and Cardiometabolic Disorders: A Review of Current Studies.
There are growing concern globally regarding the alarming trend of fast food consumption and its related cardiometabolic outcomes including overweight and obesity. This study aimed to review the current evidences available in relation to adverse effects of fast food pattern on cardiometa-bolic risk factors. Relevant articles including epidemiological and clinical studies with appropriate design and good quality were obtained through searches of the Medline, PubMed, Scopus databases and Google scholar with related key words including "fast foods", "processed foods", "obesity", "overweight", "insulin resistance", "diabetes", "cardiovascular disease", "metabolic syndrome", "dyslipidemia" and "hypertension". Fast food consumption and out-of-home eating behavior is a main risk factor for lower diet quality, higher calorie and fat intake and lower micronutrients density of diet. Frequent consumption of fast foods was accompanied with overweight and abdominal fat gain, impaired insulin and glucose homeostasis, lipid and lipoprotein disorders, induction of systemic inflammation and oxidative stress. Higher fast food consumption also increases the risk of developmental diabetes, metabolic syndrome and cardiovascular disease. This review provides further evidence warning us against the irreparable effects of fast food consumption on public health especially the increasing global burden of obesity and cardiovascu-lar diseases.


In-situ radionuclide characterization of a submarine groundwater discharge site at Kalogria Bay, Stoupa, Greece.
In-situ underwater gamma-ray spectrometer KATERINA was used for continuous measurements of radon progenies ((214)Pb, (214)Bi), thoron progeny ((208)Tl) and (40)K in submarine groundwater discharge (SGD) sites at Kalogria Bay, SW Peloponnesus (Greece). The spectrometer was deployed attached on measuring platform along with two conductivity - temperature data loggers while underwater battery packs supplied the system for acquisition periods up to 25 days. The radionuclide time series together with salinity data were obtained for spring (wet) and summer (dry) seasons. The (40)K activity concentrations correlated well with salinity of the emanating groundwater. Although the (214)Bi and (208)Tl activities showed usually similar trends anticorrelating with salinity, in some cases (208)Tl did not follow the (214)Bi record due to changes in the dynamics of the groundwater aquifer. As the half-life of (220)Rn is very short (55.6 s), its concentration in SGD may depend on the distance from its origin to the monitoring point. The observed temporal variations of (214)Bi and (208)Tl confirmed advantages of continuous in-situ monitoring of SGD in coastal areas.


Transferrin receptor expression in tumours of the human nervous system: relation to tumour type, grading and tumour growth fraction.
The expression of transferrin receptor (Tr) was investigated by means of immunohistochemistry in 101 tumours of the human central and peripheral nervous system. The results were compared with the proliferative activity of the tumours, determined by immunostaining for the proliferation-associated antigen Ki-67. In addition to immunostaining of normal and proliferated blood vessel endothelium and of a fraction of tumour infiltrating lymphocytes, we observed staining for Tr in a variable fraction of neoplastic cells of all histological tumour types. Immunoreactivity in the majority of tumour cells was found only in anaplastic tumours such as glioblastomas. Furthermore, a positive correlation between Tr expression and the Ki-67 growth fraction was established for gliomas. Non-glial tumours strongly expressing Tr included one metastatic rhabdomyosarcoma, one intracerebral malignant lymphoma, two of four plasmocytomas and seven of nine metastatic carcinomas. Our results indicate that immunohistochemistry for Tr and Ki-67 can provide additional information about the biological behaviour of nervous system tumours, thus complementing conventional histopathological criteria for anaplasia.


Sarcoidosis diagnosed in the elderly: a case-control study.
Studies on sarcoidosis in elderly patients are scarce and none have specifically evaluated patients aged ≥75 at onset. We aimed to analyse the characteristics of patients with sarcoidosis diagnosed after 75 and to compare them with those of younger patients. Multicenter case-control study comparing elderly-onset sarcoidosis (EOS) with young-onset sarcoidosis (YOS) seen at Lyon University Hospitals between 2006 and 2018. Using our institutional database, we included 34 patients in the EOS group and compared them with 102 controls from the YOS group in a 1:3 ratio. Demographic characteristics, medical history, clinical presentation, laboratory and imaging findings, sites of biopsies, histological analyses, treatments and outcomes were recorded using a comprehensive questionnaire. There were more Caucasians in the EOS group (94.1% vs. 59.8%; P < 0.001), who had significantly more comorbidities (mean, 3.1 ± 2 vs. 1.1 ± 1.6; P < 0.001). In the EOS group, there was less pulmonary involvement (26.5% vs. 49%; P = 0.022), less lymphadenopathy (2.9% vs. 16.7%; P = 0.041), no erythema nodosum (0% vs. 12.8%; P = 0.029) and no arthralgia (0% vs. 25.5%; P = 0.001). Conversely, uveitis was more common in the EOS group (55.9% vs. 20.6%; P < 0.001). Pathological confirmation was obtained significantly less frequently in the EOS group (67.7% vs. 85.3%; P = 0.023). Corticosteroid-related side effects were significantly more common in the EOS group (100% vs. 75.9%; P = 0.030). Epidemiology and clinical presentation of EOS differs from YOS, including more comorbidities and more uveitis. Elderly patients are more prone to corticosteroid side effects.


The health care experiences of British pensioners migrating to Spain: a qualitative study.
Notwithstanding the importance of cross-border healthcare mobility for Member States very little research has been carried out to understand the experiences of those that move. This paper focuses on a type of patient mobility that has had a significant growth in the past decades. These are pensioners who have moved from Northern European countries to countries in the South. To explore the health care experiences of British pensioners who have migrated to Spain. Qualitative study using in-depth interviews complemented with natural group discussions. Fifty-eight British retirees living in Spain and four that had returned to the UK from Spain. Spanish Autonomous Communities of Valencia and Baleares and the UK. British pensioners reported high satisfaction with their experiences of Spanish health care, in contrast to their more negative experiences of other public services in Spain. The perceived high quality (assessed in terms of thoroughness of care and the cleanliness of facilities) and humanity of care provided were the pivotal features of health care encounters, at all levels of the health care system. Exceptions to the overall positive assessments were transport to health care facilities, personal care in hospitals, long term care, and the management of pain relief, which were compared unfavourably with the UK NHS. In contrast to the existing literature, which suggests widespread problems faced by migrant British pensioners in accessing health care, we identified high levels of satisfaction with services.


New options and open issues in the management of unresectable stage III and in early-stage NSCLC: A report from an expert panel of Italian medical and radiation oncologists - INTERACTION group.
After the PACIFIC trial, concurrent chemo-radiotherapy followed by consolidation therapy with durvalumab for 1 year (limited to PD-L1 tumour proportion score ≥ 1% in the EMA region) is the firmly established standard of care treatment for unresectable NSCLC patients. Several relevant questions are emerging with the growing use of this approach, posing novel challenges in clinical practice. Treatment of oncogene-addicted NSCLCs, management of mediastinal disease recurrence after surgery and the optimal management of patients progressing during or after durvalumab are now some of the most clinically relevant issues. Patients with unresectable NSCLC harbouring EGFR and HER2 mutations or ALK/ROS1/RET /NTRK1,2,3 rearrangements are unresponsive to immunotherapy. Importance of knowing the tumour genotyping (NGS, preferable DNA and RNA) from the earliest stages of NSCLC, also for the possible use of immunotherapy both in the adjuvant and perioperative setting. In case of mediastinal disease recurrence after surgery, re-biopsy is essential to re-determine the histological and biological characteristics of the disease and the distinction of recurrence in curable and non-curable disease is of pivotal important for the optimal management of subsequent treatments. Treatment of stage III NSCLC has always been controversial and challenging: Multidisciplinary approach is mandatory and defining resectability is a critical issue. Chemo-radiotherapy followed by maintenance Durvalumab is now the standard of treatment. Herein, we provide a comprehensive overview of the key challenges and open questions that we are currently facing in clinical practice, in unresectable stage III and in early-stage NSCLC, identifying the knowledge gaps and the possible solutions.


Juvenile-onset bulbospinal muscular atrophy with deafness: Vialetta-van Laere syndrome or Madras-type motor neuron disease?
A girl with rapid-onset, bulbospinal muscular atrophy and deafness is described. The patient's mother and brother showed EMG features consistent with subclinical involvement. T is bulbospinal form of spinal muscular atrophy associated with deafness described by Vialetto and van Laere closely resembles the Madras type of motor neuron disease, also associated with deafness, described by Jagganathan and colleagues.


Ontogeny of cholecystokinin gene expression in the rat thalamus--a hybridization histochemical study.
We examined the ontogeny of cholecystokinin (CCK) gene expression in the rat thalamus using hybridization histochemistry. Maturation of CCK gene expression occurred over the course of a month, beginning in the medial geniculate nucleus on the 17th day of gestation. CCK mRNA was found in the caudal nuclei first, later in the rostral; the ventral were followed by the dorsal; and the lateral by the medial. In some nuclei, such as the medial geniculate, CCK mRNA was present prior to the full differentiation of the neurons; whereas in others, such as the mediodorsal, the neurons differentiated to almost adult morphology before CCK mRNA was detected.


Comparison of accuracy and precision of heart rate calibration methods to estimate total carbon dioxide production during 13C-breath tests.
13C-breath tests are noninvasive tools to measure gastrointestinal function and nutritional interventions. Calculation of percentage dose recovered of 13C in exhaled breath requires knowledge of CO2 production rate (VC02). A resting value is usually assumed, but this can underestimate VC02 because subjects are unlikely to remain at rest during tests that last for many hours. There is a need for a method to estimate nonresting VC02 during 13C-breath tests. To calibrate a heart rate monitor to continually estimate VC02 during 13C-breath tests. Proof of concept study. Eight healthy adults, 10 healthy children and six children with cystic fibrosis. Heart rate and VC02 were measured simultaneously at resting and nonresting levels. A new calibration method (smoothing heart rate and fitting a sigmoid function) was compared with published methods. A [ 3C]acetate breath test was used to demonstrate the range of physical activity during breath tests. The new calibration method was more accurate than existing methods (mean bias -0.0002%, 95% confidence interval (CI) -0.0007, 0.0003% of the mean measured VC02). Smoothing heart rate gave a more precise estimate of VC02 and a more accurate estimate of resting energy expenditure (mean bias -0.09, 95% Cl -0.22, 0.05 mmol CO2 min-' m-2 body surface area) than using raw data (mean bias -0.21, 95% Cl -0.38, -0.04 mmol CO2 min' m-2 body surface area). Physical activity level ranged from 1.0 to 2.5 in children, and 1.0 to 1.5 in adults. Use of smoothed HR with a sigmoid function provides an accurate method of estimating nonresting VC02 during 13C-breath tests.


An evaluation of the performance of five extraction methods: Chelex® 100, QIAamp® DNA Blood Mini Kit, QIAamp® DNA Investigator Kit, QIAsymphony® DNA Investigator® Kit and DNA IQ™.
DNA left at a crime scene was often limited in amount and far from pristine. To maximize the chance of recovering as much information as possible from such compromised samples, an appropriate extraction method using the available technologies needs to be devised. In this study, we used human blood, buffy coat and a total of 76 simulated touch DNA samples to test the effectiveness of the following five common DNA extraction methods, namely, Chelex® 100, QIAamp® DNA Blood Mini Kit, QIAamp® DNA Investigator Kit, QIAsymphony® DNA Investigator® Kit and DNA IQ™ system, in the recovery of such DNA. We demonstrated that the QIAamp® and QIAsymphony® DNA Investigator® Kits, and the DNA IQ™ system, exhibited a better effectiveness in DNA recovery amongst these methods and yielded extracts with higher success rate in subsequent DNA profiling. These extracts also generated profiles with better intra-colour signal balance. The findings in this work allowed us to propose an extraction approach as follows: 1) casework samples shall be extracted with the QIAamp®/QIAsymphony® DNA Investigator® Kits or the DNA IQ™ system, viz., QIAsymphony® DNA Investigator® Kit and DNA IQ™, due to their higher throughput, are for the touched DNA evidence from the volume crime, while QIAamp® DNA Investigator Kit is preferable for challenging bloodstain samples; and 2) control samples, such as buccal swab, with known identity can be extracted with the Chelex, due to their cheaper cost per sample.


Modified Children's II operation on the beating heart allows growth potential.
We describe a modification of the Children's II operation for hypoplastic left heart syndrome allowing growth potential to be performed without circulatory and cardiac arrest.


Is postoperative radiotherapy an essential treatment for nonmetastatic lymphoepithelial carcinoma of the salivary gland?
Lymphoepithelial carcinoma of salivary gland (LECSG) is a rare malignant tumor. Whether postoperative radiotherapy (PORT) can improve locoregional control and which patients can benefit from PORT are unknown. This study aimed to evaluate the role of PORT and provide individualized suggestions for postoperative therapy in patients with LECSG. We retrospectively reviewed patients with nonmetastatic LECSG who underwent surgery with or without PORT. Recursive partitioning analysis (RPA) was performed to categorize the patients and predict progression-free survival (PFS). A total of 223 patients were included, 34 (15.2%) received surgery alone, whereas the remaining 189 (84.8%) underwent PORT in the initial treatment. Although patients in the PORT group were with advanced T stage and N stage, the PORT group had an advantage over the non-PORT group on 1-year, 5-year and 10-year PFS and locoregional control (LRC). PORT was an independent prognostic factor for PFS and LRC. Furthermore, compared with T stage and N stage, the size of the primary tumor and the number of positive lymph nodes were better prognostic predictors. The RPA model was generated according to the endpoint of PFS and categorized patients into 3 prognostic groups: low-risk (maximum diameter of primary lesion (≤3 cm) and number of positive lymph nodes (≤2)), intermediate-risk (maximum diameter of primary lesion (>3 cm) and number of positive lymph nodes (≤2)), and high-risk (number of positive lymph nodes (>2)), with corresponding 5-year PFS rates of 90.0%, 75.0%, and 51.0%, respectively. Significant improvement in PFS was observed in the PORT group among intermediate-risk (P = 0.000) and high-risk patients (P = 0.000). PORT was shown to be a positive prognostic factor for PFS and LRC of LECSG. PORT was an essential treatment especially for patients with >3 cm maximum diameter of primary lesion and/or >2 positive lymph nodes.


The polycystic ovary syndrome:psychosexual correlates.
Eleven women with the polycystic ovary syndrome (POS) were interviewed prior to treatment to assess psychosexual and behavioral characteristics. Because the polycystic ovary syndrome is characterized by an elevated plasma level of free testosterone that presumably develops after birth, we examined specifically the six areas traditionally explored in prenatally androgenized women--play preference, energy expenditure level, interest in dress and appearance, career vs. family preference, sexual arousal to narrative and visual stimuli, and sexual initiative. While these women described patterns in all categories similar to those reported in the earlier studies of prenatally androgenized women, their behavior was significantly different only in sexual initiative from that of two normal groups. These findings suggest that increased sexual initiative may be a heretofore unemphasized clinical feature of the polycystic ovary syndrome and conceivably a concomitant of elevated free plasma testosterone.


Effects of ghrelin on insulin and glucagon secretion: a study of isolated pancreatic islets and intact mice.
We combined in vitro and in vivo methods to investigate the effects of ghrelin, a novel gastric hormone, on insulin and glucagon release. Studies of isolated mouse islets showed that ghrelin concentrations in the physiological range (0.5-3 nmol l(-1)) had no effect on glucose-stimulated insulin release, while low ghrelin concentrations (1-100 pmol l(-1)) inhibited and high (0.1 and 1 micromol l(-1)) stimulated. The insulin response to glucose was enhanced in the presence of a high ghrelin concentration (100 nmol l(-1)). Glucagon release was stimulated by ghrelin (0.1 pmol l(-1) to 1 micromol l(-1)); this effect was maintained in the presence of glucose (0-20 mmol l(-1)). In intact mice, basal plasma insulin was suppressed by 1 and 10 nmol kg(-1) of ghrelin, 2 and 6 min after i.v. injection. Ghrelin (0.2-10 nmol kg(-1) i.v.) suppressed also the glucose-stimulated insulin response and impaired the glucose tolerance (at a ghrelin dose of 3.3 nmol kg(-1)). Ghrelin (1 or 10 nmol kg(-1) i.v.) inhibited the insulin response to the phospholipase C stimulating agent carbachol and enhanced the insulin response to the phosphodiesterase inhibitor isobutyl-methylxanthine (IBMX) but did not affect the response to the membrane-depolarizing amino acid l-arginine. These observations suggest that the inhibitory effect of ghrelin on glucose-induced insulin release is in part exerted on phospholipase C pathways (and not on Ca(2+)entry), while the stimulatory effect of high doses of ghrelin depends on cyclic AMP. In contrast to the spectacular glucagon-releasing effect of ghrelin in vitro, ghrelin did not raise plasma glucagon. Carbachol, IBMX and l-arginine stimulated glucagon release. These responses were impaired by ghrelin, suggesting that it suppresses the various intracellular pathways (phospholipase C, cyclic AMP and Ca(2+)), that are activated by the glucagon secretagogues. Together these observations highlight (but do not explain) the different effects of ghrelin on glucagon release in vitro and in vivo. The results show that ghrelin has powerful effects on islet cells, suggesting that endogenous ghrelin may contribute to the physiological control of insulin and glucagon release. However, the narrow "window" of circulating ghrelin concentrations makes this doubtful.


Development and validation of a selective, sensitive and stability indicating UPLC-MS/MS method for rapid, simultaneous determination of six process related impurities in darunavir drug substance.
In this study a sensitive and selective gradient reverse phase UPLC-MS/MS method was developed for the simultaneous determination of six process related impurities viz., Imp-I, Imp-II, Imp-III, Imp-IV, Imp-V and Imp-VI in darunavir. The chromatographic separation was performed on Acquity UPLC BEH C18 (50 mm×2.1mm, 1.7μm) column using gradient elution of acetonitrile-methanol (80:20, v/v) and 5.0mM ammonium acetate containing 0.01% formic acid at a flow rate of 0.4mL/min. Both negative and positive electrospray ionization (ESI) modes were operated simultaneously using multiple reaction monitoring (MRM) for the quantification of all six impurities in darunavir. The developed method was fully validated following ICH guidelines with respect to specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, precision, robustness and sample solution stability. The method was able to quantitate Imp-I, Imp-IV, Imp-V at 0.3ppm and Imp-II, Imp-III, and Imp-VI at 0.2ppm with respect to 5.0mg/mL of darunavir. The calibration curves showed good linearity over the concentration range of LOQ to 250% for all six impurities. The correlation coefficient obtained was >0.9989 in all the cases. The accuracy of the method lies between 89.90% and 104.60% for all six impurities. Finally, the method has been successfully applied for three formulation batches of darunavir to determine the above mentioned impurities, however no impurity was found beyond the LOQ. This method is a good quality control tool for the trace level quantification of six process related impurities in darunavir during its synthesis.


Association of industrial work schedules with development of metabolic syndrome, insulin resistance, and serum adipokine concentrations.
Association of work schedule in industrial workers with the progression of metabolic syndrome, insulin resistance, and serum adipokine concentrations is incompletely explored. To determine the association of work schedule with the progression of metabolic syndrome, insulin resistance, and adipokine concentrations in industrial workers. In a cross-sectional study design of industrial workers we compared metabolic syndrome, insulin resistance, and adipokines concentration between workers in the day shift (n = 52), rotational shift (n = 21), and night shift (n = 15). The international Diabetes Federation criteria were used to diagnose metabolic syndrome. We used a homeostatic model assessment of insulin resistance (HOMA-IR). Serum insulin, leptin, and adiponectin concentrations were measured using enzyme-linked immunosorbent assays. Serum glucose, triglyceride, and high-density lipoprotein cholesterol (HDL-C) concentrations were monitored using Prietest clinical chemistry reagents. The proportional difference in metabolic syndrome (0.31, 95% confidence interval [CI] 0.036-0.587, P = 0.026), median difference of leptin (0.61, 95% CI 0.186-1.034, P = 0.005), and leptin-to-adiponectin ratio (LAR; 0.45, 95% CI 0.235-0.665, P < 0.001) was significantly higher, and serum adiponectin was lower (-2.00, 95% CI -4.197 to 0.197, P = 0.07) in the night-shift workers compared with that of day-shift workers. Among rotational-shift workers, the proportional difference between metabolic syndrome (0.14, 95% CI -0.098 to 0.378, P = 0.25), median difference of leptin (0.25, 95% CI -0.124 to 0.624, P = 0.19), and LAR (0.09, 95% CI -0.099 to 0.279, P = 0.35) was higher, and serum adiponectin concentration was lower (-0.73, 95% CI -2.660 to 1.208, P = 0.46) compared with that of day-shift workers; however, the altered differences were not significant. We observed a higher proportion of difference in HOMA-IR in shift workers (night and rotation) than in day-shift workers. Night-shift workers are vulnerable to a higher risk of metabolic syndrome, HOMA-IR, and adipokine changes.


Enhanced culturing techniques for the mycobiont isolated from the lichen Xanthoria parietina.
Lichens and their isolated symbionts are potentially valuable resources for biotechnological approaches. Especially mycobiont cultures that produce secondary lichen products are receiving increasing attention, but lichen mycobionts are notoriously slow-growing organisms. Sufficient biomass production often represents a limiting factor for scientific and biotechnological investigations, requiring improvement of existing culturing techniques as well as methods for non-invasive assessment of growth. Here, the effects of pH and the supplement of growth media with either D-glucose or three different sugar alcohols that commonly occur in lichens, D-arabitol, D-mannitol and ribitol, on the growth of the axenically cultured mycobiont isolated from the lichen Xanthoria parietina were tested. Either D-glucose or different sugar alcohols were offered to the fungus at different concentrations, and cumulative growth and growth rates were assessed using two-dimensional image analysis over a period of 8 weeks. The mycobiont grew at a pH range from 4.0 to 7.0, whereas no growth was observed at higher pH values. Varying the carbon source in Lilly-Barnett medium (LBM) by replacing 1% D-glucose used in the originally described LBM by either 1%, 2% or 3% of D-mannitol, or 3% of D-glucose increased fungal biomass production by up to 26%, with an exponential growth phase between 2 and 6 weeks after inoculation. In summary, we present protocols for enhanced culture conditions and non-invasive assessment of growth of axenically cultured lichen mycobionts using image analysis, which may be useful for scientific and biotechnological approaches requiring cultured lichen mycobionts. The online version contains supplementary material available at 10.1007/s11557-021-01707-7.


What the new obesity guidelines will tell us.
The purpose of the review is to update the reader on the long anticipated Obesity 2, the second iteration of 'Clinical Guidelines on the Identification, Evaluation, and Treatment of Overweight and Obesity in Adults: the Evidence Report'. Obesity 2 maintains a focus on primary care practitioners and their patients in an effort to reduce cardiovascular risk. Just as Obesity 1 limited the literature review to the years 1980-1997, Obesity 2 limits the literature under consideration from 1999 to 2011. However, because of the painstaking methodology employed, the scope of the Obesity 2 is of necessity more limited with five critical questions. These deal with risks of overweight and obesity, benefits of weight loss and evaluate three treatment areas - diet, behavioral therapies and surgical therapies. The critical questions inform recommendations in these same areas. Although not comprehensive in scope, the guidance offered in Obesity 2 is backed by stringent approaches to limit bias and assuring rigor, thus producing recommendations one can trust in the areas covered.


Avelumab vs Standard Second-Line Chemotherapy in Patients With Metastatic Colorectal Cancer and Microsatellite Instability: A Randomized Clinical Trial.
Only 1 randomized clinical trial has shown the superiority of immune checkpoint inhibitors in patients with deficient mismatch repair and/or microsatellite instability (dMMR/MSI) metastatic colorectal cancer (mCRC) in the first-line setting. To determine whether avelumab (an anti-programmed cell death ligand 1 antibody) improves progression-free survival (PFS) compared with standard second-line chemotherapy in patients with dMMR/MSI mCRC. The SAMCO-PRODIGE 54 trial is a national open-label phase 2 randomized clinical trial that was conducted from April 24, 2018, to April 29, 2021, at 49 French sites. Patients with dMMR/MSI mCRC who experienced progression while receiving standard first-line therapy were included in the analysis. Patients were randomized to receive standard second-line therapy or avelumab every 2 weeks until progression, unacceptable toxic effects, or patient refusal. The primary end point was PFS according to RECIST (Response Evaluation Criteria in Solid Tumours), version 1.1, evaluated by investigators in patients with mCRC and confirmed dMMR and MSI status who received at least 1 dose of treatment (modified intention-to-treat [mITT] population). A total of 122 patients were enrolled in the mITT population. Median age was 66 (IQR, 56-76) years, 65 patients (53.3%) were women, 100 (82.0%) had a right-sided tumor, and 52 (42.6%) had BRAF V600E-mutated tumors. There was no difference in patients and tumor characteristics between treatment groups. No new safety concerns in either group were detected, with fewer treatment-related adverse events of at least grade 3 in the avelumab group than in the chemotherapy group (20 [31.7%] vs 34 [53.1%]; P = .02). After a median follow-up of 33.3 (95% CI, 28.3-34.8) months, avelumab was superior to chemotherapy with or without targeted agents with respect to PFS (15 [24.6%] vs 5 [8.2%] among patients without progression; P = .03). Rates of PFS rates at 12 months were 31.2% (95% CI, 20.1%-42.9%) and 19.4% (95% CI, 10.6%-30.2%) in the avelumab and control groups, respectively, and 27.4% (95% CI, 16.8%-39.0%) and 9.1% (95% CI, 3.2%-18.8%) at 18 months. Objective response rates were similar in both groups (18 [29.5%] vs 16 [26.2%]; P = .45). Among patients with disease control, 18 (75.7%) in the avelumab group compared with 9 (19.1%) in the control group had ongoing disease control at 18 months. The SAMCO-PRODIGE 54 phase 2 randomized clinical trial showed, in patients with dMMR/MSI mCRC, better PFS and disease control duration with avelumab over standard second-line treatment, with a favorable safety profile. ClinicalTrials.gov Identifier: NCT03186326.


Sucrose intake lowers μ-opioid and dopamine D2/3 receptor availability in porcine brain.
Excessive sucrose consumption elicits addiction-like craving that may underpin the obesity epidemic. Opioids and dopamine mediate the rewarding effects of drugs of abuse, and of natural rewards from stimuli such as palatable food. We investigated the effects of sucrose using PET imaging with [11C]carfentanil (μ-opioid receptor agonist) and [11C]raclopride (dopamine D2/3 receptor antagonist) in seven female anesthetized Göttingen minipigs. We then gave minipigs access to sucrose solution for one hour on 12 consecutive days and performed imaging again 24 hours after the final sucrose access. In a smaller sample of five minipigs, we performed an additional [11C]carfentanil PET session after the first sucrose exposure. We calculated voxel-wise binding potentials (BPND) using the cerebellum as a region of non-displaceable binding, analyzed differences with statistical non-parametric mapping, and performed a regional analysis. After 12 days of sucrose access, BPND of both tracers had declined significantly in striatum, nucleus accumbens, thalamus, amygdala, cingulate cortex and prefrontal cortex, consistent with down-regulation of receptor densities. After a single exposure to sucrose, we found decreased binding of [11C]carfentanil in nucleus accumbens and cingulate cortex, consistent with opioid release. The lower availability of opioid and dopamine receptors may explain the addictive potential associated with intake of sucrose.


Factors affecting image magnification in indirect ophthalmoscopy with Volk or similar lenses and a biomicroscope.
To explore the factors affecting the linear magnification of the intermediate fundus image during indirect ophthalmoscopy with a slit-lamp biomicroscope. A simple paraxial model, based on a 'reduced' eye and a 'thin' ophthalmoscopy lens, is used to develop equations showing the effects of the power and ametropia of the eye, and the equivalent power and position of the ophthalmoscopy lens on fundus magnification. Predicted magnifications are compared with practical results found in earlier published experimental studies, which used Volk ophthalmoscopy lenses in conjunction with physical model eyes with adjustable levels of axial ametropia. The model's magnification predictions, as a function of the eye's ametropia, are in good agreement with previous experimental measurements, provided that the equivalent powers of the Volk lenses are used rather than their labelled nominal powers. Magnification values typically change by approximately ±10% over the practical range of each parameter if other parameters are held constant. In particular, normal variations in the equivalent power of the adult emmetropic eye result in magnifications which range from about 90-120% of the nominal value given for an eye power of +60.00 D. It is demonstrated that the recommended working distances for different powers of Volk ophthalmoscopy lenses approximate optimal matching between the various pupils of the eye-Volk lens-slit-lamp biomicroscope system. All the parameters considered have marked effects on magnification. The magnification values quoted by manufacturers can be regarded as only approximations of those which may be found in practice. Better estimates of magnification can be obtained by inserting the appropriate parameter values into the equations derived in this paper, using, where appropriate, the equivalent power of the indirect ophthalmoscopy lens, rather than the lens' labelled, 'nominal' power.


Costs and cost-effectiveness of training traditional birth attendants to reduce neonatal mortality in the Lufwanyama Neonatal Survival study (LUNESP).
The Lufwanyama Neonatal Survival Project ("LUNESP") was a cluster randomized, controlled trial that showed that training traditional birth attendants (TBAs) to perform interventions targeting birth asphyxia, hypothermia, and neonatal sepsis reduced all-cause neonatal mortality by 45%. This companion analysis was undertaken to analyze intervention costs and cost-effectiveness, and factors that might improve cost-effectiveness. We calculated LUNESP's financial and economic costs and the economic cost of implementation for a forecasted ten-year program (2011-2020). In each case, we calculated the incremental cost per death avoided and disability-adjusted life years (DALYs) averted in real 2011 US dollars. The forecasted 10-year program analysis included a base case as well as 'conservative' and 'optimistic' scenarios. Uncertainty was characterized using one-way sensitivity analyses and a multivariate probabilistic sensitivity analysis. The estimated financial and economic costs of LUNESP were $118,574 and $127,756, respectively, or $49,469 and $53,550 per year. Fixed costs accounted for nearly 90% of total costs. For the 10-year program, discounted total and annual program costs were $256,455 and $26,834 respectively; for the base case, optimistic, and conservative scenarios, the estimated cost per death avoided was $1,866, $591, and $3,024, and cost per DALY averted was $74, $24, and $120, respectively. Outcomes were robust to variations in local costs, but sensitive to variations in intervention effect size, number of births attended by TBAs, and the extent of foreign consultants' participation. Based on established guidelines, the strategy of using trained TBAs to reduce neonatal mortality was 'highly cost effective'. We strongly recommend consideration of this approach for other remote rural populations with limited access to health care.


Controversies in Dentoalveolar and Preprosthetic Surgery.
Dentoalveolar surgeries are among the more common procedures performed by oral maxillofacial surgeons. It is only natural that there are several controversies associated with many aspects of this type of surgery. Although good scientific evidence is the basis of most oral maxillofacial procedures, some of what is accepted as common wisdom may not meet strict guidelines of evidence-based practice. This article explores some controversies that are relevant to the current practice of dentoalveolar surgery.


Review article: extraoesophageal manifestations of gastro-oesophageal reflux disease.
Although recent studies suggest that gastro-oesophageal reflux disease may frequently contribute to ear, nose and throat and respiratory diseases, the cause-and-effect relationship is far from proven. The review will address this controversial topic emphasizing recent literature raising concerns about the credibility of this association and our tests to make this diagnosis. The author believes these extraoesophageal symptoms suspected to be secondary to gastro-oesophageal reflux disease are an unresolved issue, but selective use of aggressive proton-pump inhibitor therapeutic trials may help to resolve this problem in our individual patients.


γ-H2AX induced by linear alkylbenzene sulfonates is due to deoxyribonuclease-1 translocation to the nucleus via actin disruption.
Phosphorylation of histone H2AX (γ-H2AX) occurs following formation of DNA double strand breaks (DSBs). Other types of DNA damage also generate DSBs through DNA replication and repair, leading to the production of γ-H2AX. In the present study, we demonstrated that linear alkylbenzene sulfonates (LAS), the most widely used and non-genotoxic anionic surfactants, could generate γ-H2AX via a novel pathway. Breast adenocarcinoma MCF-7 cells were treated with five kinds of LAS with alkyl chains ranging from 10 to 14 carbon units (C10-C14LAS). The generation of DSBs and subsequent production of γ-H2AX increased in a manner that depended on the number of carbon units in LAS. γ-H2AX could also be generated with non-cytotoxic doses of LAS and was independent of the cell cycle, indicating the non-apoptotic and DNA replication-independent formation of DSBs. The generation of γ-H2AX could be attenuated by EGTA and ZnCl2, deoxyribonuclease-1 (DNase I) inhibitors, as well as by the knockdown of DNase I. LAS weakened the interaction between DNase I and actin, and the enhanced release of DNase I was dependent on the number of carbon units in LAS. DNase I released by the LAS treatment translocated to the nucleus, in which DNase I attacked DNA and generated γ-H2AX. These results suggested that the LAS-induced generation of γ-H2AX could be attributed to the translocation of DNase I to the nucleus through the disruption of actin, and not to LAS-induced DNA damage.


Impact of metformin use among tuberculosis close contacts with diabetes mellitus in a nationwide cohort study.
The protective effect of metformin against active tuberculosis (TB) among TB close contacts is unknown. TB close contacts with diabetes mellitus (DM) and normal renal function were selected from the National Health Insurance Research Database of Taiwan. Metformin users were patients who received ≥90 cumulative defined daily doses within 1 year before the index date. For each metformin user, a propensity-score matched metformin nonuser and an age- and sex-matched healthy TB close contact were selected. The outcome was incident TB, identified using previously validated diagnostic criteria. Independent predictors were investigated using stratified Cox regression analysis. Interaction analysis was also performed. A total of 5846 TB close contacts who were metformin users, metformin non-users, and healthy contacts were analysed. The incidence of active TB was 755 (600-938), 1117 (927-1335), and 526 (393-689) cases per 100,000 person-years in each group, respectively. Multivariate analysis revealed that healthy contacts had the lowest risk of developing active TB (adjusted hazard ratio [aHR]: 0.42 [0.30-0.60]) and metformin use partially reversed the risk associated with DM (aHR: 0.73 [0.54-0.98]). Subpopulation analysis revealed a significant interaction between insulin use and metformin use. Metformin use is associated with a lower risk of developing active TB among TB close contacts with DM, especially for insulin users. It may be an alternative choice for primary prevention against active TB if no contraindications exist. However, prospective studies are needed to confirm the findings.


A correction score to compare aortic calcification in contrast enhanced and non-contrast measurements from computed tomography scans.
Aortic wall calcification shows strong promise as a cardiovascular risk factor. While useful for visual enhancement of vascular tissue, enhancement creates heterogeneity between scans with and without contrast. We evaluated the relationship between aortic calcification in routine abdominal computed tomography scans (CT) with and without contrast. Inclusion was limited to those with abdominal CT-scans with and without contrast enhancement within 120 days. Analytic Morphomics, a semi-automated computational image processing system, was used to provide standardized, granular, anatomically indexed measurements of aortic wall calcification from abdominal CT-scans. Aortic calcification area (ACA) and aortic wall calcification percent (ACP) and were the outcomes of interest. Multiple linear regression was used to evaluate the relationship of aortic measurements. Models were further controlled for age and sex. Stratification of measurements by vertebral level was also performed. A positive association was observed for non-contrast calcification in ACP β 0.74 (95% CI 0.72, 0.76) and ACA β 0.44 (95% 0.43, 0.45). Stratified results demonstrated the highest coefficient of determination at L2 for percent and L3 for area models [R2 0.91 (ACP) 0.74 (ACA)]. Adjusted lumber-level associations between non-contrast and contrast measurements ranged from (β 0.69-0.82) in ACP and (β 0.37-0.54) in ACA. A straightforward correction score for comparison of abdominal aortic calcification measurements in contrast-enhanced and non-contrast scans is discussed. Correction of aortic calcification from CT scans can reduce scan heterogeneity and will be instrumental in creating larger cardiovascular cohorts as well as cardiovascular risk surveillance programs.


[The securing of perfusion nozzles for hemosorption in flasks with various capacities].
A device for the fastening of a standard perfusion nozzle on flasks with thread-free necks for hemosorption is described. The device enables to perform hemosorption procedures when it is not appropriate to use standard 400-ml flasks.


Preterm birth alters the feeding-induced activation of Akt signaling in the muscle of neonatal piglets.
Postnatal lean mass accretion is commonly reduced in preterm infants. This study investigated mechanisms involved in the blunted feeding-induced activation of Akt in the skeletal muscle of preterm pigs that contributes to lower protein synthesis rates. On day 3 following cesarean section, preterm and term piglets were fasted or fed an enteral meal. Activation of Akt signaling pathways in skeletal muscle was determined. Akt1 and Akt2, but not Akt3, phosphorylation were lower in the skeletal muscle of preterm than in term pigs (P < 0.05). Activation of Akt-positive regulators, PDK1 and mTORC2, but not FAK, were lower in preterm than in term (P < 0.05). The formation of Akt complexes with GAPDH and Hsp90 and the abundance of Ubl4A were lower in preterm than in term (P < 0.05). The abundance of Akt inhibitors, PHLPP and SHIP2, but not PTEN and IP6K1, were higher in preterm than in term pigs (P < 0.05). PP2A activation was inhibited by feeding in term but not in preterm pigs (P < 0.05). Our results suggest that preterm birth impairs regulatory components involved in Akt activation, thereby limiting the anabolic response to feeding. This anabolic resistance likely contributes to the reduced lean accretion following preterm birth. The Akt-mTORC1 pathway plays an important role in the regulation of skeletal muscle protein synthesis in neonates. This is the first evidence to demonstrate that, following preterm birth, the postprandial activation of positive regulators of Akt in the skeletal muscle is reduced, whereas the activation of negative regulators of Akt is enhanced. This anabolic resistance of Akt signaling in response to feeding likely contributes to the reduced accretion of lean mass in premature infants. These results may provide potential novel molecular targets for intervention to enhance lean growth in preterm neonates.


Organization of the rat UDP-glucuronosyltransferase, UDPGTr-2, gene and characterization of its promoter.
A lambda clone containing the entire gene and flanking sequences for a form of UDP-glucuronosyltransferase (UDPGTr-2) that glucuronidates testosterone and the foreign compounds, 4-hydroxybiphenyl and chloramphenicol, has been isolated from a rat liver genomic library. Sequence analysis of this clone revealed that the UDPGTr-2 gene is approximately 12 kilobase pairs in length and consists of six exons. All introns were found to interrupt protein coding regions of the gene. Four transcriptional start sites have been identified and are located 34, 35, 38, and 39 base pairs (bp) upstream of the translation initiation site. The 5'-flanking region of the gene contains a TATA-like sequence, CATAAA, 22 bp from the first transcription start site, potential AP-1 and v-MYB binding sites, and four sequence motifs that have been found in genes that are expressed predominantly in the liver. A 323-bp fragment encompassing these elements was fused upstream in both orientations to the coding sequence of placental alkaline phosphatase to assay promoter activity. Transient transfection of various cultured cell lines with the chimeric DNA demonstrated that this fragment, in the correct orientation, was able to function as an efficient promoter in the rat hepatoma cell lines Reuber H4-II-E and McA-RH7777. It was, however, inactive in hepatoma cell lines from two other species and in cell lines derived from other tissues. These results are consistent with the physiological expression of the rat UDPGTr-2 gene and suggest that the proximal 5'-flanking region of the gene may contain information which limits its expression to the liver.


A recombinant rabies virus carrying GFP between N and P affects viral transcription in vitro.
Several studies have demonstrated the rabies virus to be a perfect potential vaccine vector to insert foreign genes into the target genome. For this study, a green fluorescent protein (GFP) gene was cloned into the rabies virus (RABV) genome between the N and P gene. CT dinucleotide was inserted as intergenic region. The recombinant high egg passage Flury strain (HEP-Flury) of RABV, carrying GFP (rHEP-NP-GFP), was generated in BHK-21 cells using reverse genetics. According to the viral growth kinetics assay, the addition of GFP between N and P gene has little effect on the viral growth compared to the parental strain HEP-Flury. Quantitative real-time PCR (qPCR) indicated that rHEP-NP-GFP showed different viral gene transcription, especially for G gene, compared to HEP-Flury. The same is true for one other recombinant RABV carrying GFP between G and L gene in NA cells. In addition, parent HEP-Flury showed more expression of innate immune-related molecules in NA cells. Compared to HEP-Flury, Western blotting (WB) indicated that insertion of a foreign gene following N gene enhanced the expression of M and G proteins. According to the qPCR and WB, GFP expression levels of rHEP-NP-GFP were significantly higher than rHEP-GFP. This study indicates HEP-Flury as valid vector to express exogenous genes between N and P.


Endothelium-specific insulin resistance leads to accelerated atherosclerosis in areas with disturbed flow patterns: a role for reactive oxygen species.
Systemic insulin resistance is associated with a portfolio of risk factors for atherosclerosis development. We sought to determine whether insulin resistance specifically at the level of the endothelium promotes atherosclerosis and to examine the potential involvement of reactive oxygen species. We cross-bred mice expressing a dominant negative mutant human insulin receptor specifically in the endothelium (ESMIRO) with ApoE(-/-) mice to examine the effect of endothelium-specific insulin resistance on atherosclerosis. ApoE(-/-)/ESMIRO mice had similar blood pressure, plasma lipids and whole-body glucose tolerance, but blunted endothelial insulin signalling, in comparison to ApoE(-/-) mice. Atherosclerosis was significantly increased in ApoE(-/-)/ESMIRO mice at the aortic sinus (226 ± 16 versus 149 ± 24 × 10(3) μm(2), P = 0.01) and lesser curvature of the aortic arch (12.4 ± 1.2% versus 9.4 ± 0.9%, P = 0.035). Relaxation to acetylcholine was blunted in aorta from ApoE(-/-)/ESMIRO mice (Emax 65 ± 41% versus 103 ± 6%, P = 0.02) and was restored by the superoxide dismutase mimetic MnTMPyP (Emax 112 ± 15% versus 65 ± 41%, P = 0.048). Basal generation of superoxide was increased 1.55 fold (P = 0.01) in endothelial cells from ApoE(-/-)/ESMIRO mice and was inhibited by the NADPH oxidase inhibitor gp91ds-tat (-12 ± 0.04%, P = 0.04), the NO synthase inhibitor L-NMMA (-8 ± 0.02%, P = 0.001) and the mitochondrial specific inhibitor rotenone (-23 ± 0.04%, P = 0.006). Insulin resistance specifically at the level of the endothelium leads to acceleration of atherosclerosis in areas with disturbed flow patterns such as the aortic sinus and the lesser curvature of the aorta. We have identified a potential role for increased generation of reactive oxygen species from multiple enzymatic sources in promoting atherosclerosis in this setting.


Intracellular localization of Prostatic Binding Protein (PBP) in rat prostate by light and electron microscopic immunocytochemistry.
Extra- and intracellular distribution of Prostatic Binding Protein (PBP) was studied in the different genital organs of the male rat by immunocytochemistry at the light and electron microscopic levels. PBP was extracted from cytosols of rat ventral prostate and used for immunization of rabbits. The specificity of the antiserum raised was tested by "western blotting" and immunoelectrophoresis. From the different fixatives tested for optimal structural and antigenic preservation of the ventral prostate a mixture containing 2.5% paraformaldehyde, 0.5% glutaraldehyde and 0.5% CaCl2 in cacodylate buffer, 0.05 M, pH 7.3 was selected. Using the immunofluorescence technique and the unlabeled antibody enzyme method PBP-immunoreactivity was detected at the light microscopic level in the luminal secretions of the ventral prostate. No reaction was observed with the seminal vesicle, the coagulating gland, the dorsal and lateral prostates, the epididymis and the testis. Intracellular secretory granules reacting with PBP antiserum were exclusively found in the secretory cells of the ventral prostate. Insufficiently fixed cells showed a diffuse generalized reaction of the cytoplasm indicating a leakage of the antigen from the secretory granules. Such artifacts were common in tissue sections processed with the preembedding-staining procedure. At the ultrastructural level therefore mostly the postembedding staining method was performed using both the unlabeled antibody enzyme method and the ferritin-labeled immunoglobulin technique in osmicated, Epon-embedded tissue. Labeling with either method was intense in the secretory granules and the condensing vacuoles, while the labeling density of the rough endoplasmic reticulum and the Golgi cisternae was in the background range. Castration experiments showed that secretory material displaying PBP immunoreactivity was retained within the acinar lumen of the gland for several days after castration, but was absent from most secretory cells already by four days after castration. Immunocytochemistry of PBP therefore is a very sensitive method for analysing the secretory activity and its androgen dependence of the prostate of the rat.


Insulin uptake by rat liver endothelium studied in fractionated liver cell suspensions.
Cellular distribution of insulin receptors was studied in fractionated rat liver cell suspensions using 125I-insulin and a visual probe consisting of latex beads covalently linked to insulin (minibeads). Fractionation was done on metrizamide gradients which yielded two cellular fractions. The large cell fraction consisted mostly of hepatocytes and the small cell fraction consisted of 37% endothelial cells as well as Kupffer cells. The magnitude of insulin uptake by the endothelium-rich small cell fraction was at least double that of the uptake by the hepatocyte-rich fraction. The minibead technique demonstrated that in the small cell fraction only endothelial cells, and not Kupffer cells, were responsible for the insulin uptake. Our findings suggest that liver endothelium may be responsible for the uptake of circulating insulin and its transport to hepatocyte. This emphasizes the presence of a tissue-blood barrier in the liver.


Investigating the impact of in situ soil organic matter degradation through porewater spectroscopic analyses on marsh edge erosion.
Marsh edge erosion results in soil organic matter (SOM) loss from coastal wetlands and is differentially affected by wind waves, soil properties, and vegetation cover. The degradation of SOM may make the marsh edge susceptible to erosion. The objective of this study was to investigate the effect of in situ biogeochemical degradations of SOM on marsh edge erosion using porewater spectroscopic analyses. Edge erosion was monitored at 12 transects in one of the highly eroding coastal basins of Louisiana. A total of 36 cores were collected at different distances from the edge of the marsh. Porewater was extracted and analyzed for dissolved organic carbon (DOC) and spectroscopic indicators. The north and west side had greater erosion rates (102.38 ± 5.2 cm yr-1) than east and south side (78.47 ± 3.3 cm yr-1). However, the north and east side had greater DOC and refractory carbon but less microbial activity indicating SOM degradation alone did not correlate to edge erosion. The intersecting trend between erosion rate and SOM degradation among four sides of the island indicates the complex nature of edge erosion drivers. The estuarine bottom indicators suggest the eroded SOM is not reburied but rather degraded and emitted back into the atmosphere as CO2, potentially contributing to global change. The coastlines projected to experience high sea-level rise in the coming century are vulnerable to losing a large amount of stored carbon in the absence of efficient mitigation measures.


Facial solitary morphea profunda presenting with painful trigeminal neuropathy: A case report.
Localized facial scleroderma usually presents as frontal linear morphea or progressive hemifacial atrophy. Only isolated cases of trigeminal painful neuropathy have been described. A 43-year-old woman developed an oval lesion on the right cheek. After 1 year, she noticed constant "pulling" pain and episodes of lancinating pain, both spontaneous and triggered by chewing and cold drinks. She was diagnosed with solitary morphea profunda and CT scan, ultrasonography, cranial MRI and biopsy were completed. Methylprednisolone (1 gr/day for 3 days) was prescribed. For pain, gabapentin, oxcarbazepine, amitryptiline, pregabalin and eslicarbacepine were all ineffective. A capsaicin patch was placed with prolonged benefit. Later on, the pain slightly worsened; occipital blockade was effective and methotrexate was recommended. This is the first case of solitary morphea profunda associated with painful trigeminal neuropathy. Treatment should include immunosuppressants and treatment of neuropathic pain, in which local therapies seem particularly beneficial.


Plunging ranula: a report of three cases and review of the literature.
Three cases of plunging ranula are reported and the literature reviewed. Extravasation of saliva from the sublingual gland due to trauma or obstruction of its ducts appears to be the most likely cause of plunging ranula. Available data suggest that the submandibular gland is usually not involved, although at the time of surgery it may be extremely difficult to exclude a submandibular origin of the cyst in the neck. Communication between the oral and cervical components of the plunging ranula probably occur via a hiatus in the mylohyoid muscle. Such communication passing directly into the submandibular compartment may simulate submandibular gland involvement. Since 1910, 139 procedures in 89 patients with plunging ranula have been reported in the English literature. The recurrence rate was 70 per cent after incision and drainage of the cyst, 53 per cent after marsupialization, 85 per cent after excision of the cyst in the neck and 2 per cent after excision of the sublingual gland via the cervical or intra-oral route. This review suggests that excision of the sublingual gland with intra-oral drainage of the cervical swelling appears to be the treatment of choice for the plunging ranula.


Improving drying kinetics, physicochemical properties and bioactive compounds of red dragon fruit (Hylocereus species) by novel infrared drying.
Effects of tray rotation speeds (TRS: 0, 20, 40 rpm), temperatures (50, 60, 70 °C) and wavelength spectra (mid and near-infrared) were comparatively evaluated on improving drying kinetics, physicochemical properties and bioactive content of red dragon fruits. Results indicated that successive increases in TRS and temperature led to significant reductions in drying time and increases in drying rates and moisture diffusivity. High TRS (40 rpm) and lower temperatures (50, 60 °C) also improved colour, total soluble solids, rehydration ratio, total phenolics and flavonoid contents, betalain content and antioxidant activity. Meanwhile, NIR drying presented a more energy-efficient approach, but with substantial reductions in quality properties compared with MIR drying. Overall, the results suggested the importance of wavelength absorption properties of plant tissues and potential avoidance of localized overheating for enhanced efficiency during infrared drying and prompted the development of suitable approaches and optimization studies for improving efficiency.


Simultaneous quantitation of twenty-two amino acids in nanolitre samples of biological fluid.
The technique of thin-layer chromatography and 14C-dansylation has been used for simultaneous measurements of 22 individual amino acids, taurine, serotonin, and gamma-aminobutyric acid in nanolitre samples of glomerular and renal tubule fluid. Standard curves of each amino acid mixture containing all the others showed excellent linearity at amounts ranging from 5.7.10(-13) to 1.45.10(-11) mole (regression coefficients all greater than 0.95). With careful standardization of dansylation conditions, the technique permits highly reproducible measurement of less than 1 pmole of an amino acid.


Sandwich-ELISE NMR: reducing the sample volume of NMR samples.
We present Sandwich-ELISE, a concatenated version of our previously proposed Experimental LIquid SEaling (ELISE) protocol, in which an aqueous sample is effectively sealed by the addition of a small layer of mineral oil, or, alternatively, a chloroform sample was sealed by a water layer. With Sandwich-ELISE, a triple layered geometry composed of deuterated chloroform/aqueous buffer/mineral oil can be used to limit the sample to the active coil volume, effectively replacing the popular Shigemi tubes. Importantly, this procedure is readily applicable to smaller diameter tubes, for which no Shigemi tubes are available. We further present spectra of a 1 microl protein sample sandwiched between the chloroform and Nujol phases in a 1mm tube, demonstrating thereby that the volume of the aqueous phase of interest can be reduced even further.


Salvage chemotherapy according to the ASHAP protocol: a single-center study of 24 patients with relapsed or refractory aggressive non-Hodgkin's lymphomas.
Numerous salvage protocols for relapsed or refractory aggressive non-Hodgkin's lymphomas have been described. The purpose of this retrospective study was to evaluate the efficacy and the toxicity of the ASHAP protocol, which combines a continuous infusion of doxorubicin and cisplatin with high-dose cytarabine and methylprednisolone. Twenty-four patients with relapsed or refractory aggressive non-Hodgkin's lymphomas were treated with a median of 3 cycles (range: 1-5) of ASHAP. Twelve patients achieved a complete and four a partial remission for an overall response rate of 67%. The 3-year overall and progression-free survival rates were 60% and 40%, respectively. Ten of the responding patients were consolidated by high-dose chemotherapy. After a median follow-up of 15.5 months, four patients are in continuous complete remission, while six patients suffered relapses (two fatal). For reasons of low risk profile [international prognostic index (IPI) score of 0, n=2] or age >60 years ( n=4), consolidation was limited to involved-field radiotherapy in six patients. All of these patients are alive after a median follow-up of 37 months, with two relapses. Factors predicting a poor response to salvage therapy were primary refractory disease, elevated lactate dehydrogenase activity, and an IPI score of >/=2. The principal toxicity was myelosuppression with grade III or IV neutropenia or thrombocytopenia occurring in 88% or 75%, respectively, of the patients. Nonhematological toxicity was generally mild. There were no treatment-related deaths. The ASHAP regimen is a highly active and well-tolerated salvage protocol for patients with relapsed aggressive non-Hodgkin's lymphomas which compares favorably with other established protocols.


Choosing the location for non-image guided abdominal paracentesis.
The optimal location for paracentesis has not been studied scientifically. The evolving obesity epidemic has changed the physique of many patients with cirrhosis and ascites such that needles inserted into the abdominal wall may not reach fluid. We aimed to determine the location for paracentesis that would have the thinnest abdominal wall and the deepest amount of fluid. Ultrasound measurements of abdominal wall thickness and depth of ascites were recorded in two locations, the infraumbilical midline (ML) and the left lower quadrant (LLQ), in 52 patients with cirrhosis and ascites admitted to a single inpatient liver unit. The abdominal wall was significantly thinner (1.8 vs. 2.4 cm; P<0.001) and the depth of ascites greater (2.86 vs. 2.29 cm; P=0.017) in the LLQ as compared with the infraumbilical ML position. In the left lateral oblique position, the difference in the depth of ascites was more pronounced when comparing the LLQ with the infraumbilical ML (4.57 vs. 2.78 cm; P<0.0001). The LLQ is preferable to the ML infraumbilical location for performing paracentesis.


Fetal calf serum and retinoic acid affect proliferation and terminal differentiation of a rat rhabdomyosarcoma cell line (BA-HAN-1C).
We report on the establishment of a model for differentiation induction in sarcomas, using the clonal rhabdomyosarcoma cell line BA-HAN-1C. This rhabdomyosarcoma cell line is composed of morphologically undifferentiated mononuclear stem cells, some of which spontaneously fuse to form terminally differentiated multinuclear myotube-like giant cells. The deprivation of fetal calf serum (FCS) or the exposure to retinoic acid, respectively, resulted in a significant inhibition of proliferation (P less than 0.001) and a marked increase in cellular differentiation as shown by a significant increase in the number of myotube-like giant cells (P less than 0.001) and in the creatine kinase activity (P less than 0.05) used as a biochemical marker of differentiation. Furthermore, after exposure to retinoic acid about 30% of the mononuclear tumour cells exhibited morphological features of rhabdomyogenic differentiation, such as bundles of thick and thin myofilaments, which had never been observed in the mononuclear cells of untreated cultures. These results confirm that the inverse linkage between proliferation and differentiation known from embryonic myogenesis is preserved in our rhabdomyosarcoma cell line. The failure to induce terminal differentiation by exposure to retinoic acid in all the cells of our clonal cell line indicates that some tumour cells might epigenetically be blocked from responding to retinoic acid. The temporary growth retardation observed after FCS-deprivation suggests that autocrine stimulation of proliferation may be operating in our cell line, too.


Associative-memory representations emerge as shared spatial patterns of theta activity spanning the primate temporal cortex.
Highly localized neuronal spikes in primate temporal cortex can encode associative memory; however, whether memory formation involves area-wide reorganization of ensemble activity, which often accompanies rhythmicity, or just local microcircuit-level plasticity, remains elusive. Using high-density electrocorticography, we capture local-field potentials spanning the monkey temporal lobes, and show that the visual pair-association (PA) memory is encoded in spatial patterns of theta activity in areas TE, 36, and, partially, in the parahippocampal cortex, but not in the entorhinal cortex. The theta patterns elicited by learned paired associates are distinct between pairs, but similar within pairs. This pattern similarity, emerging through novel PA learning, allows a machine-learning decoder trained on theta patterns elicited by a particular visual item to correctly predict the identity of those elicited by its paired associate. Our results suggest that the formation and sharing of widespread cortical theta patterns via learning-induced reorganization are involved in the mechanisms of associative memory representation.


Minimal change nephrotic syndrome four days after the administration of Pfizer-BioNTech COVID-19 vaccine-a new side effect or coincidence?
The association between the COVID vaccine and MCD is temporal and by exclusion, and it is not fully established, but it should be considered in postvvaccine MCD.


[Research on the Combined Detection of Urine UmAlb and Urinary Nephrin, Podocalyxin in Podocyte of MKR Mice with Diabetic Nephropathy].
To explore the combined detection of urine UmAlb and urinary nephrin (Unephrin), podocalyxin (UPCX) in podocyte of MKR mice with diabetic nephropathy. Thirty 8 weeks old MKR mice were randomly divided into two groups as follows: negative control group, DN model group, and another 15 wild C57 mice were used as normal control. Mice in DN model group were received unilateral nephrectomy and high-fat diet feed for 2 months. The morphological structure changes of the podocytes were observed by transmission electron microscopes. The levels of FBG were detected by electrochemical detection method, The nephrin and PCX protein expression were measured by western blotting. The levels of UmAlb, Unephrin and UPCX were detected by ELISA. The podocyte damage in the mice of DN model group increased significantly when compared with normal control. As compared with normal control, FBG in the model group increased significantly (P<0. 01), the expression level of nephrin and PCX in Renal Tissue and Unephrin, UPCX, and urine UmAlb were also increased significantly (P<0. 01). The level of Unephrin and UPCX were positive correlated with the level of urine UmAlb, the loss of podocyte strcture protein might be one of the mechanism in leading proteinuria in diabetic nephropathy.


Scuba diving accidents in Sweden 1960--1976.
Thirty "scuba" diving accidents (29 of them fatal) which occurred in Sweden during 1960--76 are listed. Nine of these are described in some detail. The probable causes of these accidents are evaluated, and suggestions for prophylactic measures are made.


Variables in the measurement of serum cholesterol by gas-liquid chromatography.
Gas-liquid chromatography (GLC) is an effective method for the measurement of total serum cholesterol. Its use in the clinical laboratory, however, requires careful attention to many variables (flow rate, sample size, sensitivity of solvent and choice of the quantitation method, i.e. peak-height ratio vs. peak height). This report evaluates the effect of these variables on the quantitation of serum cholesterol by GLC.


Behavioral and ERP measures of holistic face processing in a composite task.
Holistic processing of faces is characterized by encoding of the face as a single stimulus. This study employed a composite face task to examine whether holistic processing varies when attention is restricted to the top as compared to the bottom half of the face, and whether evidence of holistic processing would be observed in event-related potentials. Analyses of behavioral data showed that spatial misalignment of the face halves disrupted holistic processing and enhanced detection of repeated attended halves. Effects of misalignment on the N170, VPP and N250 ERP components resembled effects of face inversion. Attention to the top half of the face was associated with faster P1, N170, VPP, and P2 latencies than attending to the bottom, suggesting automatic processing of the eye region. Further, N170 latency effects suggested that structural encoding of the face is facilitated during holistic processing. N250 latency effects reflected task difficulty. Finally, an overall right hemispheric asymmetry was most pronounced when holistic face processing was greatest. Results are discussed in light of recent proposals that holistic face processing is a subtype of configural face processing.


Evaluation of an Oral Hygiene Education Program for Staff Providing Long-Term Care Services: A Mixed Methods Study.
Oral hygiene is often neglected in clients receiving long-term care, suggesting that long-term care workers require formal oral hygiene education. Thus, the aim of this study was to investigate the effects of oral hygiene education on long-term care workers. This study utilized a mixed methods design. Eighty long-term care workers were recruited for participation in the oral hygiene education program, which employed three teaching methods: narration with multimedia presentation, demonstration, and teach-back. The effect of the education program on the participants' level of oral hygiene knowledge, attitudes, and skills was measured using a structured questionnaire that was administered both pre- and post-delivery of the education program. Three months later, all participants submitted a self-report of their oral hygiene skills, and six participants completed a telephone interview. Quantitative data were analyzed using paired t-tests, and qualitative data were manually analyzed and coded. Scores of oral hygiene knowledge (p < 0.001), attitudes (p = 0.001), and oral cleaning daily frequency for clients (p < 0.001), were significantly higher three months after undertaking the educational program. This preliminary study suggests that oral hygiene education may be effective in improving oral hygiene knowledge, attitudes, and skills among long-term care staff.


In vivo forces generated by finger flexor muscles do not depend on the rate of fingertip loading during an isometric task.
Risk factors for activity-related tendon disorders of the hand include applied force, duration, and rate of loading. Understanding the relationship between external loading conditions and internal tendon forces can elucidate their role in injury and rehabilitation. The goal of this investigation is to determine whether the rate of force applied at the fingertip affects in vivo forces in the flexor digitorum profundus (FDP) tendon and the flexor digitorum superficialis (FDS) tendon during an isometric task. Tendon forces, recorded with buckle force transducers, and fingertip forces were simultaneously measured during open carpal tunnel surgery as subjects (N=15) increased their fingertip force from 0 to 15N in 1, 3, and 10s. The rates of 1.5, 5, and 15N/s did not significantly affect FDP or FDS tendon to fingertip force ratios. For the same applied fingertip force, the FDP tendon generated more force than the FDS. The mean FDP to fingertip ratio was 2.4+/-0.7 while the FDS to tip ratio averaged 1.5+/-1.0 (p<0.01). The fine motor control needed to generate isometric force ramps at these specific loading rates probably required similar high activation levels of multiple finger muscles in order to stabilize the finger and control joint torques at the force rates studied. Therefore, for this task, no additional increase in muscle force was observed at higher rates. These findings suggest that for high precision, isometric pinch maneuvers under static finger conditions, tendon forces are independent of loading rate.


A fatal infection caused by sequence type 398 methicillin-resistant Staphylococcus aureus carrying the Panton-Valentine leukocidin gene: A case report in Japan.
Methicillin-resistant Staphylococcus aureus (MRSA) has now been recognized as a common pathogen in the community. Sequence type (ST) 398 MRSA is generally considered as an emerging zoonotic agent spreading among livestock and personnel who have direct contact with animals, mainly in Europe. A 37-year-old Chinese woman receiving steroid therapy for systemic lupus erythematosus with general fatigue and myalgia was brought to the emergency department in critical condition. Her condition deteriorated despite aggressive management and she died on day 7. Her blood culture revealed ST398 MRSA-SCCmec V with Panton-Valentine Leukocidin (PVL) gene. This is the first case report of a fatal infection caused by this lineage. According to the results of molecular analyses, the isolate from this particular patient's blood was genetically close to a lineage detected in China, and is less likely to be related to an animal-associated lineage.


Five-year trends for ventilator-associated pneumonia: Correlation between microbiological findings and antimicrobial drug consumption.
The epidemiology of multidrug-resistant bacteria (MDRB) has changed significantly in European healthcare settings, with a decrease in frequency of meticillin-resistant Staphylococcus aureus and an increase in extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae. Little is known about the effects of these changes on ventilator-associated pneumonia (VAP). A retrospective 5-year trend analysis of ICU antibiotic consumption and resistance in bacteria causing VAP was undertaken. Poisson regression analysis between complete microbiological data and antibiotic consumption was performed. In total, 252 episodes of VAP in 184 patients were identified between 2007 and 2011, from which 364 causal bacteria were isolated. Enterobacteriaceae isolation rates increased significantly over this period [from 6.64 to 10.52 isolates/1000 patient-days; P=0.006], mostly due to an increase in AmpC-producing Enterobacteriaceae (APE) (2.85-4.51 isolates/1000 patient-days; P=0.013), whereas the number of episodes due to S. aureus and Pseudomonas aeruginosa remained stable. A positive association was found between the increase in APE infections and an increase in past-year antibiotic consumption: amoxicillin/clavulanic acid (P=0.003), ceftazidime and cefepime (P=0.007), carbapenems (P=0.002), fluoroquinolones (P=0.012), macrolides (P=0.002) and imidazoles (P=0.004). No such association was found for the emergence of resistance in P. aeruginosa. These results indicate a change in the epidemiology of VAP, with Enterobacteriaceae exceeding P. aeruginosa and S. aureus. Moreover, a positive correlation was observed between antibiotic consumption and the incidence of potentially MDRB such as APE. No such correlation was found for ESBL-producing Escherichia coli and antibiotic-resistant P. aeruginosa.


Concentrating Autographa californica nuclear polyhedrosis virus and recombinant alkaline phosphatase from insect cells using a temperature-sensitive hydrogel.
Recombinant alkaline phosphatase expressed in insect cells was concentrated by a factor of one and half times at a separation efficiency of 54.2% using hydrogel ultrafiltration. Enzyme concentration was confirmed by SDS-PAGE as well as by spectrophotometric measurement. Wild and recombinant Autographa californica nuclear polyhedrosis viruses (AcNPV) were concentrated 1.4 and 1.6 times of the feed solution at 48.5 and 60.0% separation efficiency, respectively. Hydrogel ultrafiltration appears to be an attractive alternative for the concentration of AcNPV and recombinant proteins from insect cells.


A-412997, a selective dopamine D4 agonist, improves cognitive performance in rats.
The recent development of a highly selective dopamine D4 receptor agonist, A-412997 (2-(3',4',5',6'-tetrahydro-2'H-[2,4'] bipyridinyl-1'-yl)-N-m-tolyl-acetamide), has provided a pharmacological tool with which to conduct systematic investigations into the putative role for dopamine D4 receptors in the central nervous system. These present studies evaluated the potential cognitive enhancing properties of A-412997 in rat models of ADHD (5-trial repeated acquisition inhibitory avoidance in Spontaneous Hypertensive Rat pups) and short-term memory (Social Recognition), in comparison with the less selective dopamine D4 receptor agonists PD168077 and CP226269. A-412997 showed significant dose-dependent efficacy in both models. PD168077 repeatedly improved acquisition in the 5-trial inhibitory avoidance model but failed to reach significance at any dose tested, although significantly improved social recognition was observed (albeit less potent than A-412997). CP226269 showed a significant enhancement in the 5-trial inhibitory avoidance model. These results support a role for the dopamine D4 receptor subtype in cognition.


mRNA COVID-19 vaccines are well tolerated and myopericarditis is a rare adverse event following immunisation.
mRNA vaccines are considered to be important tools for the management of the COVID-19 pandemic. Although mRNA COVID-19 vaccines are well tolerated in most recipients, post-marketing data have highlighted an association between myocarditis and mRNA vaccines. Post-vaccine myocarditis (PVM) is most commonly reported in male adolescents receiving the second dose of an mRNA vaccine. However, the incidence of PVM is low and the risk of myocarditis should be kept in perspective. Cases of PVM are mostly mild in severity, and may be managed using existing myocarditis guidelines. The pathology of PVM is under investigation, and current data suggest that cross reactivity or hypersensitivity reactions may be involved. Globally, mRNA vaccines are generally recommended for use in children/adolescents, and delaying the administration of the second dose may reduce the risk of PVM.


Is surgical indication in multiple myeloma a poor prognosis sign? SEER database analysis.
This study aims to investigate demographic data, survival rates, and the relationship of these rates with surgery in a large case series including multiple myeloma (MM) patients. MM cases were analyzed retrospectively using the latest version of the SEER database published in April 2020. This version covers January 1975 to December 2017. Patients were classified according to gender, age, and race/ethnicity. Tumors were classified according to their localization, grade, year of diagnosis, and follow-up results. There were 60,239 patients diagnosed with Plasma Cell Myeloma. While 670 patients (1.2%) were operated on, 43,976 patients (76.7%) did not indicate operation, and 12,670 patients (22.1%) could not be operated on despite the recommendation. The mean survival was 62 months in those without an indication for surgery, and 42 months in patients with an indication but could not be operated on, and the difference was significant (p = 0.001). The mean survival was 58 months in the operated patients, and 42 months in the patients who could not be operated on despite the indication, and the difference was significant (p = 0.001). There was no difference between those who did not indicate surgery and those who were operated on with an indication (p = 0.243). In multiple myeloma, the best prognosis is in the group of patients who received medical treatments without any indication for operation, while an indication for operation indicates a worse prognosis. A worse prognosis should be expected in patients who do not accept the operation or who cannot be operated on compared to the operated patients.


Tumor-infiltrating and circulating granulocytic myeloid-derived suppressor cells correlate with disease activity and adverse clinical outcomes in mycosis fungoides.
Cutaneous T cell lymphomas (CTCL) are rare and histologically diverse lymphoproliferative neoplasms, with mycosis fungoides (MF) representing the most common disease subset. Given the emerging role of myeloid-derived suppressor cells (MDSC) as a clinically applicable biomarker in solid tumors, we sought to investigate the presence of tumor-infiltrating and circulating MDSC in early- and advanced-stage MF patients and evaluate their prognostic significance in patient overall survival. Tumor-infiltrating MDSC were assessed immunohistochemically with Arginase-1 in 31 MF and 14 non-MF skin punch biopsies. Circulating MDSC were assessed with flow cytometry in freshly isolated PBMC from 29 MF patients. Granulocytic MDSC (G-MDSC) were defined as CD11b+CD14-CD15+ and monocytic MDSC (M-MDSC) were defined as CD11b+CD14+HLA-DRlow/-. MDSC infiltration occurred in approximately one-third (35.5%) of CTCL lesions, with a predilection for non-MF lesions (p < 0.05). The predominant morphology of MDSC was granulocytic. Although in MF lesions the presence of MDSC infiltrates did not correlate with clinical stage, it conferred significantly worse overall survival outcomes (p < 0.05). Circulating G-MDSC were significantly higher in MF patients compared to healthy donor controls (p < 0.0001), while M-MDSC did not show any statistically significant difference. G-MDSC were significantly higher in patients with active disease compared to patients who were in partial remission (p < 0.01). As with tumor-infiltrating MDSC, clinical stage did not correlate with circulating G-MDSC levels, while prospective overall survival analysis showed that patients with high levels of circulating G-MDSC have significantly inferior outcomes (p < 0.01). This study shows that G-MDSC could represent a novel and easily assessable biomarker in MF, which mirrors disease activity and can predict patient subgroups with aggressive clinical features.


Surveying the elimination of hazardous heavy metal from the multi-component systems using various sorbents: a review.
In this review, several adsorbents were studied for the elimination of heavy metal ions from multi-component wastewaters. These utilized sorbents are mineral materials, microbes, waste materials, and polymers. It was attempted to probe the structure and chemistry characteristics such as surface morphology, main functional groups, participated elements, surface area, and the adsorbent charges by SEM, FTIR, EDX, and BET tests. The uptake efficiency for metal ions, reusability studies, isotherm models, and kinetic relations for recognizing the adsorbent potentials. Besides, the influential factors such as acidity, initial concentration, time, and heat degree were investigated for selecting the optimum operating conditions in each of the adsorbents. According to the results, polymers especially chitosan, have displayed a higher adsorption capacity relative to the other common adsorbents owing to the excellent surface area and more functional groups such as amine, hydroxyl, and carboxyl species. The high surface area generates the possible active sites for trapping the particles, and the more effective functional groups can complex more metal ions from the polluted water. Also, it was observed that the uptake capacity of each metal ion in the multi-component solutions was different because the ionic radii of each metal ion were different, which influence the competition of metal ions for filling the active sites. Finally, the reusability of the polymers was suitable, because they can use several cycles which proves the economic aspect of the polymers as the adsorbent.


Diminished foveal sensitivity may predict the development of advanced age-related macular degeneration.
Visual function testing was performed on one eye with drusen from each of 18 elderly patients in 1984. Eleven patients had advanced age-related macular degeneration (AMD) in the fellow eye, and seven had only bilateral drusen. These patients were all followed prospectively (median, 45 months), at which time one eye had a new vessel membrane, three eyes had pigment epithelial detachments, and one eye had geographic atrophy. Only two of these five eyes had AMD-related visual loss in the fellow eye in 1984. The degree of loss of foveal dark-adapted sensitivity in 1984 predicted which patients developed advanced AMD with 100% sensitivity and 92% specificity. The presence of high-risk drusen characteristics in 1984 predicted the development of advanced AMD with 100% sensitivity but only 55% specificity. For this small group of patients, foveal dark-adapted sensitivity loss was an excellent predictor of the subsequent development of advanced AMD in eyes with drusen. A prospective study of a large group of patients with drusen is being undertaken to validate this finding.