Metabolic networks in a porcine model of trauma and hemorrhagic shock demonstrate different control mechanism with carbohydrate pre-feed.
Treatment with oral carbohydrate prior to trauma and hemorrhage confers a survival benefit in small animal models. The impact of fed states on survival in traumatically injured humans is unknown. This work uses regulatory networks to examine the effect of carbohydrate pre-feeding on metabolic response to polytrauma and hemorrhagic shock in a clinically-relevant large animal model. Male Yorkshire pigs were fasted overnight (n = 64). Pre-fed animals (n = 32) received an oral bolus of Karo\textregistered\syrup before sedation. All animals underwent a standardized trauma, hemorrhage, and resuscitation protocol. Serum samples were obtained at set timepoints. Proton NMR was used to identify and quantify serum metabolites. Metabolic regulatory networks were constructed from metabolite concentrations and rates of change in those concentrations to identify controlled nodes and controlling nodes of the network. Oral carbohydrate pre-treatment was not associated with survival benefit. Six metabolites were identified as controlled nodes in both groups: adenosine, cytidine, glycerol, hypoxanthine, lactate, and uridine. Distinct groups of controlling nodes were associated with controlled nodes; however, the composition of these groups depended on feeding status. A common metabolic output, typically associated with injury and hypoxia, results from trauma and hemorrhagic shock. However, this output is directed by different metabolic inputs depending upon the feeding status of the subject. Nodes of the network that are related to mortality can potentially be manipulated for therapeutic effect; however, these nodes differ depending upon feeding status.


Transcriptional implications of intragenic DNA methylation in the oestrogen receptor alpha gene in breast cancer cells and tissues.
DNA methylation variability regions (MVRs) across the oestrogen receptor alpha (ESR1) gene have been identified in peripheral blood cells from breast cancer patients and healthy individuals. In contrast to promoter methylation, gene body methylation may be important in maintaining active transcription. This study aimed to assess MVRs in ESR1 in breast cancer cell lines, tumour biopsies and exfoliated epithelial cells from expressed breast milk (EBM), to determine their significance for ESR1 transcription. DNA methylation levels in eight MVRs across ESR1 were assessed by pyrosequencing bisulphite-converted DNA from three oestrogen receptor (ER)-positive and three ER-negative breast cancer cell lines. DNA methylation and expression were assessed following treatment with DAC (1 μM), or DMSO (controls). ESR1 methylation levels were also assayed in DNA from 155 invasive ductal carcinoma biopsies provided by the Breast Cancer Campaign Tissue Bank, and validated with DNA methylation profiles from the TCGA breast tumours (n = 356 ER-pos, n = 109 ER-neg). DNA methylation was profiled in exfoliated breast epithelial cells from EBM using the Illumina 450 K (n = 36) and pyrosequencing in a further 53 donor samples. ESR1 mRNA levels were measured by qRT-PCR. We show that ER-positive cell lines had unmethylated ESR1 promoter regions and highly methylated intragenic regions (median, 80.45%) while ER-negative cells had methylated promoters and lower intragenic methylation levels (median, 38.62%). DAC treatment increased ESR1 expression in ER-negative cells, but significantly reduced methylation and expression of ESR1 in ER-positive cells. The ESR1 promoter was unmethylated in breast tumour biopsies with high levels of intragenic methylation, independent of ER status. However, ESR1 methylation in the strongly ER-positive EBM DNA samples were very similar to ER-positive tumour cell lines. DAC treatment inhibited ESR1 transcription in cells with an unmethylated ESR1 promoter and reduced intragenic DNA methylation. Intragenic methylation levels correlated with ESR1 expression in homogenous cell populations (cell lines and exfoliated primary breast epithelial cells), but not in heterogeneous tumour biopsies, highlighting the significant differences between the in vivo tumour microenvironment and individual homogenous cell types. These findings emphasise the need for care when choosing material for epigenetic research and highlights the presence of aberrant intragenic methylation levels in tumour tissue.


A short history of the initial discovery of the Wolf-Hirschhorn syndrome.
Deletion of the short arm of chromosome 4 (4p-) was first described in 1961 [Hirschhorn and Cooper, 1961], and the second case of 4p- was published in 1965 [Hirschhorn et al., 1965]. This short history describes the original case and the sequence of events leading to the publications.


Detection of human papillomavirus-16 in ovarian malignancy.
Human papillomavirus is the causal factor for cervical cancer. However, the role of HPV infection in ovarian cancer is unclear. This study aimed to determine the presence of human papillomavirus-16 (HPV-16) in ovarian cancer tissues. Archived human ovarian cancer tissues (N=54 cases, 50 are epithelial cancer, four are nonepithelial cancer) embedded in paraffin blocks were used. Controls are 30 nonmalignant ovarian tissue blocks. In situ hybridisation (ISH) and immunohistochemistry (IHC) were used to detect the presence of HPV-16 and p53 expression. In all, 52 or 36% of the epithelial ovarian tumours detected by ISH or IHC, respectively, were HPV-16 E6 positive. In contrast, only 6.7% of normal ovarian tissues were HPV-16 positive proved by ISH. Human papillomavirus-16 infection was significantly higher in cancer tissues compared to controls with an odds ratio of 16.7 (95% confidence interval [CI]=3.2-71.4, P<0.01). No significant correlation between HPV-16 infection and histological types of cancer was found (P>0.05). p53 gene expression was detected in 42% epithelial ovarian cancers. No correlation between p53 expression and HPV-16 infection was found. The results showed the presence of HPV-16 E6 in ovarian carcinoma, suggesting that HPV infection might play a role in ovarian carcinogenesis.


Predictive factors for central lymph node metastasis in patients with cN0 papillary thyroid carcinoma: A systematic review and meta-analysis.
Recently, some single-institution studies have reported risk factors of CLNM in cN0 PTC patients, but results from those studies are not consistent. The meta-analysis aimed to identify some risk factors that can be predictive of CLNM in cN0 PTC patients. We performed a systematic search for relevant literature published prior to December 2015 using the following search engines: PubMed, EMBASE, Ovid and Web of Science. We included retrospective, prospective, and observational studies that investigated the risk factors for CLNM in patients with cN0 PTC. Thirty-one studies, including 37,355 patients with cN0 PTC from seven countries were included in the meta-analysis. The pooled analysis indicated that age<45 years (OR = 1.57, 95%CI:1.48-1.66, P < 0.001), male gender (OR = 1.79, 95%CI: 1.69-1.91, P < 0.001), tumor size>10 mm (OR = 2.61, 95%CI:2.27-3.00, P < 0.001), bilaterality (OR = 1.52, 95%CI:1.31-1.77, P < 0.001), multifocality (OR = 1.46, 95%CI: 1.31-1.61, P < 0.001), extracapsular invasion (OR = 2.10, 95%CI:1.81-2.43, P < 0.001), angiolymphatic invasion (OR = 8.02, 95%CI:5.00-12.87, P < 0.001), high histologic risk (OR = 2.62, 95%CI:2.13-3.22, P < 0.001) and BRAF(V600E) mutation (OR:1.78, 95%CI:1.38-2.30, P < 0.001) were significantly associated with CLNM, and upper third location (OR = 0.54, 95%CI:0.43-0.67, P < 0.001) and lymphocytic thyroiditis (OR = 0.64, 95%CI:0.42-0.97, P = 0.034) were decreased risk factors of CLNM. We identified several predictive factors for CLNM in cN0 PTC patients. Some certain risk factors could be considered in preoperative clinical decision regarding the necessity of prophylactic central lymph node dissection in cN0 PTC patients.


The association between vitamin D levels in the second trimester of pregnancy and gestational diabetes mellitus.
25-hydroxyvitamin D (25(OH)D) affects glucose metabolism by increasing insulin secretion and insulin receptor expression. However, whether 25(OH)D deficiency will increase the risk of gestational diabetes mellitus (GDM) has not been clearly reported. The purpose of this study is to assess the relationship between vitamin D levels in the second trimester of pregnancy and the risk of GDM. According to the inclusion and exclusion criteria, 247 pregnant women came to the fourth hospital of Shijiazhuang (The affiliated obstetrics and gynecology hospital of Hebei Medical University) for obstetrics were investigated during the period of January 1, 2019 to December 31, 2020. The levels of 25(OH)D in the second trimester (16-20 weeks) and oral 75 g glucose tolerance test (OGTT) at 24-28 weeks of pregnancy were reviewed. The sociodemographic data were collected from questionnaire. Multivariate logistic regression was used to analyze the relationship between vitamin D levels and GDM. The incidence of GDM in the observation group (25(OH)D ≤ 26 ng/ml) was higher than that in the control group (25(OH)D > 26 ng/ml) (p = 0.039). Compared with control group, the observation group had significantly higher level of fasting plasma glucose (FPG) (4.7 [4.5-5.0] mmol/L vs. 4.6 [4.4-4.8] mmol/L, p = 0.012). In the whole study, the level of 25(OH)D was negatively correlated with FPG (r = - 0.164,p = 0.010). After adjusting for age, pre-pregnancy BMI, parity and adverse pregnancy history, compared with the observation group (25 (OH) D ≤ 26 ng/ml), the risk of developing GDM decreased by 50.9% in control group (25(OH)D > 26 ng/ml) (odds ratio [OR] = 0.491, 95% confidence interval [CI] = 0.243-0.989, p = 0.047). Adequate vitamin D levels during the second trimester of pregnancy may reduce the risk of GDM.


Duodenocolic fistula: an unusual complication of duodenal diverticulum.
The management of two patients with benign duodenocolic fistula is reported. In both patients, laparotomy clearly demonstrated the origin of a fistulous tract arising from a duodenal diverticulum. Microscopic examination of the diverticula failed to disclose the presence of either gastric or pancreatic tissue.


Transitioning the critical care nurse from ICU to high-tech homecare.
Critical care nurses are in a good position to take advantage of the changes in health care. They can build a career that will be on the cutting edge of the changes in technology and care delivery if they are willing to learn some new skills and practice in a more autonomous environment. Nurses who are new to the homecare setting need to know more about insurance systems, diagnosis coding, and outcomes tools and prepare themselves for the rapid growth of computer assisted technologies that are being introduced into the practice of homecare. They also need to take the whole continuum of care into consideration when planning care for patients in the home and to be aware of community and family supports that can enhance the patient's health related quality of life.


Antioxidants in health and disease: overview.
Molecular oxygen is an essential nutrient for higher forms of life. In addition to its normal physiological reactions, oxygen and its partially reduced forms can oxidize a variety of macromolecular and simpler compounds in cells and fluids of the body. Such oxidized and peroxidized compounds have been associated with, and may be causally related to, a variety of chronic diseases. As a protection against excessive oxidation, nature has developed a complex set of interactive antioxidant systems. Selected aspects of antioxidant actions are considered in this symposium.


Separation of fibronectin from a plasma gelatinase using immobilized metal affinity chromatography.
Conventional preparations of plasma fibronectin are known to contain a co-purifying gelatinase [1986, J. Biol. Chem. 261, 4363-4366], but so far useful methods to remove the protease have not been available. In this study a number of different methods were tested in order to achieve separation of the two proteins. Immobilized metal affinity chromatography was found to be efficient for this purpose, and a convenient procedure to separate the two proteins under nondenaturing conditions on chelating Sepharose charged with Co2+, Ni2+, or Zn2+ is described. An alternative method employing pH gradient elution of an Fe3+ gel also resolved fibronectin from the gelatinase. The Fe3+ gel bound both proteins at pH 6.0 but not at pH 7.4, suggesting that the two proteins were phosphorylated. The described procedures will now allow studies of the functions of fibronectin in the absence of the contaminating protease.


Concomitant Use of Quinolones and Stimulants and the Risk of Adverse Cardiovascular Symptoms: A Retrospective Cohort Study.
To examine whether concomitant use of quinolones and stimulants increases the risk of cardiac events in adults. A retrospective cohort study of privately insured adults using MarketScan® claims data from 2008 to 2015. Stimulant (methylphenidate or mixed amphetamine salts) users (18-65 yrs old) with continuous health plan enrollment for the 6 months (baseline) prior to the first dispensation (index date) of oral quinolones or comparators (amoxicillin ± clavulanate or azithromycin). OUTCOMES DEFINITION: (1) Cardiac symptoms (palpitation, tachycardia, or syncope); (2) cardiac arrhythmias (ventricular arrhythmias, paroxysmal ventricular tachycardia, or cardiac arrest). Baseline covariates adjustment was through inverse probability of treatment weighting. Adults were followed until the antimicrobial therapy ended. The hazard of cardiac events in stimulant-quinolones-exposed adults was compared to those who were treated with stimulant-comparator antibiotics using a weighted Cox regression model. Several sensitivity analyses were performed to challenge the results robustness. The study cohorts comprised 390,490 stimulants users who initiated either quinolone or amoxicillin, and 387,574 patients receiving stimulants who initiated quinolone or azithromycin. The unadjusted incidence rate for cardiac symptoms in stimulant-quinolones users was 471 cases/10,000 patient-years, and it was 244 cases/10,000 patient-years in patients exposed to stimulant-amoxicillin; whereas the unadjusted incidence rate for cardiac symptoms was 728 and 358 per 10,000 patient-years for stimulant-quinolones and stimulant-azithromycin cohorts, respectively. Compared to stimulant-amoxicillin use, the adjusted hazard ratio (HR) for cardiac symptoms with stimulant-quinolones use was 1.61 (95% confidence interval [CI], 1.30-1.98). The HR for cardiac symptoms for patient exposed to stimulant-quinolones was 1.69 (95% CI, 1.32-2.13) when compared to stimulant-azithromycin. The sensitivity analysis findings were consistent with the primary analysis. A few patients across the study comparison groups developed cardiac arrhythmias. Concomitant use of stimulants and quinolone was associated with an increased hazard of cardiac symptoms in comparison to concomitant use of stimulants and amoxicillin or azithromycin, but there was no apparent difference in cardiac arrhythmias.


Non-operative management of shoulder osteoarthritis: Current concepts.
Glenohumeral osteoarthritis (OA) is one of the most common causes of shoulder pain. Conservative treatment options include physical therapy, pharmacological therapy, and biological therapy. Patients with glenohumeral OA present shoulder pain and decreased shoulder range of motion (ROM). Abnormal scapular motion is also seen in patients as adaptation to the restricted glenohumeral motion. Physical therapy is performed to (1) decrease pain, (2) increase shoulder ROM, and (3) protect the glenohumeral joint. To decrease pain, it should be assessed whether the pain appears at rest or during shoulder motion. Physical therapy may be effective for motion pain rather than rest pain. To increase shoulder ROM, the soft tissues responsible for the ROM loss need to be identified and targeted for intervention. To protect the glenohumeral joint, rotator cuff strengthening exercises are recommended. Administration of pharmacological agents is the major part next to physical therapy in the conservative treatment. The main aim of pharmacological treatment is the reduction of pain and diminution of inflammation in the joint. To achieve this aim, non-steroidal anti-inflammatory drugs are recommended as first-line therapy. Additionally, the supplementation of oral vitamin C and vitamin D can help to slow down cartilage degeneration. Depending on the individual comorbidities and contraindications, sufficient medication with good pain reduction is thus possible for each patient. This interrupts the chronic inflammatory state in the joint and, in turn, enables pain-free physical therapy. Biologics such as platelet-rich plasma, bone marrow aspirate concentrate, and mesenchymal stem cells have gathered increased attention. Good clinical outcomes have been reported, but we need to be aware that these options are helpful in decreasing shoulder pain but neither stopping the progression nor improving OA. Further evidence of biologics needs to be obtained to determine their effectiveness. In athletes, a combined approach of activity modification and physical therapy can be effective. Oral medications can provide patients with transient pain relief. Intra-articular corticosteroid injection, which provides longer-term effects, must be used cautiously in athletes. There is mixed evidence for the efficacy of hyaluronic acid injections. There is still limited evidence regarding the use of biologics.


Lipoprotein abnormalities associated with lipopolysaccharide-induced lecithin: cholesterol acyltransferase and lipase deficiency.
Density gradient ultracentrifugation was used to isolate and characterize the plasma lipoproteins from African green monkeys before and 24 and 48 h after subcutaneous injection of 300 micrograms/kg lipopolysaccharide (LPS) to induce an acute phase response. Compared with 0 h values, reductions occurred in plasma cholesterol (39%), high density lipoprotein (HDL) cholesterol (54%), lecithin:cholesterol acyltransferase (LCAT) activity (55%), and post-heparin plasma lipase activity (68%) 48 h after LPS injection while plasma triglyceride concentrations increased 700%. Cholesterol distribution among lipoproteins shifted from 7 to 41% in very low density lipoproteins (VLDL), 65 to 38% in low density lipoproteins (LDL), and 28 to 21% in HDL after LPS injection. At 48 h after LPS injection, all lipoprotein classes were relatively enriched in phospholipid and triglyceride and depleted of cholesteryl ester. The plasma concentration of all chemical constituents in VLDL was increased 3-9-fold within 48 h after LPS injection. By negative stain electron microscopy, HDL were discoidal in shape while VLDL and LDL appeared to have excess surface material present. Even though total HDL protein concentration in plasma was unaffected, the plasma mass of the smallest HDL subfractions (HDL3b,c) doubled while the mass of intermediate-sized subfractions (HDL3a) was dramatically decreased within 24 h after treatment. HDL became enriched in apoE, acquired apoSAA, and became depleted of apoA-I, A-II, and Cs by 48 h after LPS injection while apoB-100 remained the major apoprotein of VLDL and LDL. We conclude that administration of LPS to monkeys prevents normal intravascular metabolism of lipoproteins and results in the accumulation of relatively nascent forms of lipoproteins in plasma. These immature lipoproteins resemble those isolated from the recirculating perfusion of African green monkey livers, which are relatively deficient of LCAT activity and those isolated from the plasma of patients with familial LCAT deficiency.


Smartphone-based thermography to determine shunt patency in patients with hydrocephalus.
When shunt dysfunction is suspected, radiation exposure due to X-rays or a CT-scan is inevitable. Less-invasive and more reliable methods are warranted. In this study, we aimed to assess the usability of smartphone-based thermography to detect shunt patency in patients with hydrocephalus. This prospective observational pilot study evaluated the use of smartphone-based video thermography to detect flow of cerebrospinal fluid in the shunt of 51 patients from the Department of Neurosurgery at a tertiary referral institute. Patients with a shunt for hydrocephalus without the suspect of dysfunction were included in the study from December 2021 to May 2022. We included 51 patients with a mean age of 53.3 years. Of these patients 14 were male (27.5%) and 37 were female (72.5%). The most frequent cause of the hydrocephalus was the normal pressure hydrocephalus followed by the congenital hydrocephalus. Most patients (96%) had a ventriculoperitoneal shunt, whereas two had a ventriculo-atrial shunt. In total, 43 patient (84%) had a shunt on the right side and 8 patient (16%) had the shunt located on the left side. In 45 patients (88.2%), we observed a clear flow of cerebrospinal fluid in the cooled shunt trajectory. The findings of this study indicate that in patients with a shunt to treat hydrocephalus, the smartphone-based video thermography may be a safe and simple alternative to show shunt patency without the exposure to radiation.


Involvement of agmatine in antidepressant-like effect of HMG-CoA reductase inhibitors in mice.
3-Hydroxy-3-methyl-glutaryl-co-enzyme-A (HMG-CoA) reductase inhibitors (statins) are popularly used for the treatment of obesity and hypercholesterolemia with established safety profile. Statins exhibits a wide range of neurobehavioral effects in addition to their peripheral actions, and may be beneficial in treatment of psychiatric conditions. Present study investigated the role of agmatine and imidazoline receptors in antidepressant-like effect of statins in mouse forced swimming test (FST). The antidepressant-like effect of atorvastatin (5 mg/kg, p.o.) and simvastatin (10 mg/kg, p.o.) was potentiated by pretreatment with agmatine (5 mg/kg, i.p.) as well as the drugs known to increase endogenous agmatine levels in brain viz., L-arginine (40 μg/mouse, i.c.v.), an agmatine biosynthetic precursor; arcaine (50 μg/mouse, i.c.v), agmatinase inhibitor; and aminoguanidine (6.5 μg/mouse, i.c.v.), a diamine oxidase inhibitor. Further, both the statins increased agmatine levels within hippocampus and prefrontal cortex. Conversely, prior administration of I1 receptor antagonist, efaroxan (1 mg/kg, i.p.) and I2 receptor antagonist, idazoxan (0.25 mg/kg, i.p.) blocked the antidepressant-like effect of statins and their synergistic combination with agmatine. These results demonstrate the involvement of agmatine and imidazoline receptors in antidepressant-like effect of statins and suggest as a potential therapeutic target for the treatment of depressive disorders.


Membrane fusion of enveloped viruses: especially a matter of proteins.
To infect mammalian cells, enveloped viruses have to deposit their nucleocapsids into the cytoplasm of a host cell. Membrane fusion represents a key element in this entry mechanism. The fusion activity resides in specific, virally encoded membrane glycoproteins. Some molecular properties of these fusion proteins will be briefly described. These properties will then be correlated to the ability of a virus to fuse with target membranes, and to induce cell-cell fusion. Some molecular and physical parameters affecting virus fusion--at the level of either viral or target membrane or both--and the significance of modelling virus fusion by using synthetic peptides resembling viral fusion peptides, will also be discussed.


The structure of the large spacer region of the rDNA in Vicia faba and Pisum sativum.
The complete nucleotide sequence of the rDNA large spacer region was determined for two related legume plants, Vicia faba and Pisum sativum. In the large spacers of both plants, there were one or several tandem arrays of repeated sequences which account well for the length heterogeneity of the rDNA repeating units. When the entire lengths of Vicia and Pisum rDNA large spacers were compared, significant sequence similarity ranging from approximately 50 to 80% was observed, thus a close phylogenetical relationship was revealed. However, organization patterns of these rDNA spacers were clearly different from each other. The Pisum rDNA large spacer contained only one type of repeated sequence of about 175 bp, whereas the Vicia contained five kinds of repeated sequence. In addition, the initiation site for transcription was tentatively determined by S1 protection experiment.


Association of medial collateral ligament complex injuries with anterior cruciate ligament ruptures based on posterolateral tibial plateau injuries.
The combined injury of the medial collateral ligament complex and the anterior cruciate ligament (ACL) is the most common two ligament injury of the knee. Additional injuries to the medial capsuloligamentous structures are associated with rotational instability and a high failure rate of ACL reconstruction. The study aimed to analyze the specific pattern of medial injuries and their associated risk factors, with the goal of enabling early diagnosis and initiating appropriate therapeutic interventions, if necessary. Between January 2017 and December 2018, 151 patients with acute ACL ruptures with a mean age of 32 ± 12 years were included in this study. The MRIs performed during the acute phase were analyzed by four independent investigators-two radiologists and two orthopedic surgeons. The trauma impact on the posterolateral tibial plateau and associated injuries to the medial complex (POL, dMCL, and sMCL) were examined and revealed an injury to the medial collateral ligament complex in 34.4% of the patients. The dMCL was the most frequently injured structure (92.2%). A dMCL injury was significantly associated with an increase in trauma severity at the posterolateral tibial plateau (p < 0.02) and additional injuries to the sMCL (OR 4.702, 95% CL 1.3-133.3, p = 0.03) and POL (OR 20.818, 95% CL 5.9-84.4, p < 0.0001). Isolated injuries to the sMCL were not observed. Significant risk factors for acquiring an sMCL injury were age (p < 0.01) and injury to the lateral meniscus (p < 0.01). In about one-third of acute ACL ruptures the medial collateral ligament complex is also injured. This might be associated with an increased knee laxity as well as anteromedial rotational instability. Also, this might be associated with an increased risk for failure of revision ACL reconstruction. In addition, we show risk factors and predictors that point to an injury of medial structures and facilitate their diagnosis. This should help physicians and surgeons to precisely diagnose and to assess its scope in order to initiate proper therapies. With this in mind, we would like to draw attention to a frequently occurring combination injury, the so-called "unlucky triad" (ACL, MCL, and lateral meniscus). Level of evidence Level III Retrospective cohort study.


Fuzzy monitoring of in-bed postural changes for the prevention of pressure ulcers using inertial sensors attached to clothing.
Postural changes while maintaining a correct body position are the most efficient method of preventing pressure ulcers. However, executing a protocol of postural changes over a long period of time is an arduous task for caregivers. To address this problem, we propose a fuzzy monitoring system for postural changes which recognizes in-bed postures by means of micro inertial sensors attached to patients' clothes. First, we integrate a data-driven model to classify in-bed postures from the micro inertial sensors which are located in the socks and t-shirt of the patient. Second, a knowledge-based fuzzy model computes the priority of postural changes for body zones based on expert-defined protocols. Results show encouraging performance in the classification of in-bed postures and high adaptability of the knowledge-based fuzzy approach.


Is renin substrate an erythropoietin precursor?
The biogenesis of erythropoietin is incompletely understood. One hypothesis maintains that erythropoietin is synthesized primarily in the kidney while according to another theory an erythropoietin precursor present in plasma is activated by a renal factor, erythrogenin. An attractive candidate for the erythropoietin precursor is renin substrate (angiotensinogen) which has chemical similarities with erythropoietin. We show here that purified renin substrate from human plasma is immunologically related to human erythropoietin. Moreover, purified renin substrate, like erythropoietin, causes the dose-dependent increase of haemoglobin F in cultured human erythroid leukaemia K562 cells. We conclude that renin substrate is a likely precursor of erythropoietin.


Clearance of influenza virus infections by T cells: risk of collateral damage?
Influenza A viruses are a major cause of respiratory infections in humans. To protect against influenza, vaccines mainly aim at the induction of antibodies against the two surface proteins and do not protect against influenza A viruses from other subtypes. There is an increasing interest in heterosubtypic immunity that does protect against different subtypes. CD8 and CD4 T cells have a beneficial effect on the course of influenza A virus infection and can recognize conserved IAV epitopes. The T cell responses are tightly regulated to avoid collateral damage due to overreaction. Different studies have shown that an aberrant T cell response to an influenza virus infection could be harmful and could contribute to immunopathology. Here we discuss the recent findings on the balance between the beneficial and detrimental effects of T cell responses in influenza virus infections.


[Prevalence of symptoms associated with blood pressure in normal and hypertensive population].
High blood pressure should be symptomless. However, popular knowledge attributes some symptoms such as headache, epistaxis or dizziness to high blood pressure. To compare the prevalence of headache, epistaxis, subconjunctival hemorrhages and dizziness in people with normal or high blood pressure. A representative sample of an urban population in Chile was studied. Each subject with high blood pressure was paired with a non-related individual of the same sex and age. These subjects were interrogated about the presence of symptoms. One thousand forty eight subjects with high blood pressure (481 male) and 1052 individuals with normal pressure (483 male), were studied. In subjects with high or normal blood pressure, the prevalence of headache was 22 and 21.8% respectively, epistaxis was 11.6 and 11% respectively, dizziness was 7.4 and 7.6% respectively, tinnitus was 9.1 and 9.4% respectively and subconjunctival hemorrhage was 2.9 and 3.1% respectively (p = NS). No differences in the prevalence of symptoms, popularly attributed to hypertension, were observed between subjects with normal or high blood pressure.


Alterations in aortic endothelial cell morphology and cytoskeletal protein synthesis during cyclic tensional deformation.
Previous studies have shown that bovine aortic endothelial cells (ECs) in culture respond to repetitive tensional deformation with an increase in deoxyribonucleic acid synthesis and cell proliferation. This study was designed to determine whether cyclic tensional deformation of ECs in vitro induces different morphologic or protein synthetic responses. ECs from passages 6 through 9 were seeded in 35 mm2 well silicone rubber plates at 2 x 10(5) cells/well and allowed to attach for 24 hours. The experimental group was placed in a vacuum-operated stress-providing device that exerted an elongation of 24% at maximum downward deflection of the culture plate bottom and was subjected to repetitive cycles of 10 seconds of 24% maximum elongation and 10 seconds of relaxation for 5 days. The control group was subjected to similar incubation conditions but without stretch. 35S-methionine (500 muCi/well) was added to the plates 24 hours before harvesting, and two-dimensional gels of the harvested lysates (isoelectric focusing with pH 3 to 10 ampholytes followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 12.5% gels) were performed and the labeled proteins visualized by autoradiography. The data indicate that there is a differential synthesis of proteins, with synthesis of some proteins decreased or ablated whereas other proteins were increased in response to cyclic mechanical tension. The actin filament organization was evaluated after staining with rhodamine phalloidin, a fluorescent F-actin probe. The ECs subjected to tension had a more polygonal shape and demonstrated pseudopods and actin stress fibers, whereas ECs cultured under static conditions were more rounded and did not express actin stress cables.(ABSTRACT TRUNCATED AT 250 WORDS)


Bone and skeletal muscle metastases from gastric adenocarcinoma: unusual radiographic, CT and scintigraphic features.
Skeletal metastatic lesions arising from gastric cancer are uncommon and usually of the osteolytic type. In order to gain a better understanding of its radiological spectrum, we report two atypical cases of skeletal metastases from gastric adenocarcinoma presenting with unusual radiographic, CT and scintigraphic features. In one patient multiple ossifying skeletal muscle metastases and bone metastases with spiculated periosteal reaction occurred as a presenting manifestation of the malignant disease. The other patient developed widespread osteosclerotic metastases with a superscan pattern on bone scintigraphy.


Associations between cardiac function and retinal microvascular geometry among Chinese adults.
Abnormal retinal microvascular geometry has been associated with cardiac remodeling and heart failure. However, its relation to cardiac function, prior to clinical disease has not been explored. In this cross-sectional study, 50 participants (mean age 62.5 ± 11.7 years) without cardiovascular disease (CVD) were recruited from the Cardiac Ageing Study. Transthoracic echocardiography imaging was performed to measure cardiac function indices, and retinal imaging was used to measure retinal vascular caliber and retinal vascular geometric indices. Multiple linear regressions were applied to examine associations between indices of cardiac function and retinal microvasculature, adjusting for age, sex, body mass index, mean blood pressure and comorbidity (i.e. hypertension, diabetes and dyslipidemia). After adjusting for confounders, each unit decrease in peak systolic septal mitral annular velocity (Septal S') indicating poorer left function was associated with smaller retinal venular branching angle (β: - 2.69°; 95% CI - 4.92, - 0.46). Furthermore, each unit increase in peak velocity flow in late diastole by atrial contraction (MV A Peak) indicating poorer left atrial function was associated with lower retinal venular fractal dimension (- 0.13Df; - 0.25, - 0.004). Our findings suggested a relationship between poorer cardiac function and suboptimal retinal microvascular geometry, among Chinese without CVD.


PsbS genotype in relation to coordinated function of PS II and PS I in Arabidopsis leaves.
Application of multiple probes to systems that carry specific mutations provides a powerful means for studying how known regulators of light utilization interact in vivo. Two lines of Arabidopsis thaliana were studied, each carrying a unique lesion in the nuclear psbS gene encoding a 22-kDa pigment-binding protein (PS II-S) essential for full expression of photoprotective, rapid-phase, nonphotochemical quenching of chlorophyll fluorescence (NPQ). The PS II-S protein is absent in line npq4-1 due to deletion of psbS. Line npq4-9 expresses normal levels of PS II-S but carries a single amino acid substitution that lowers NPQ capacity by about 50%. A prior report [Peterson RB and Havir EA (2001) Planta 214: 142-152] described an altered pattern of redox states of the acceptor side of Photosystem II (PS II) and donor side of Photosystem I (PS I) for npq4-9 suggesting that interphotosystem electron transport may be restricted by a higher transthylakoid DeltapH in this line. In vivo steady state fluorescence and absorbance measurements (820 nm) confirmed these earlier observations for line npq4-9 but not for npq4-1. Thus, the prior results cannot be correlated simply to a loss of NPQ capacity. Likewise, the kinetics of the 820-nm absorbance change did not indicate a substantial effect of psbS genotype on electron flow from plastoquinol to PS I. A simple model is proposed to relate linear electron transport rate (measured gasometrically) to a parameter (based on fluorescence) that provides a relative measure of the density of excitation available for photochemistry in PS II. Surprisingly, analyses using this model suggested that the in vivo midpoint potential of the primary quinone acceptor in PS II (Q(A)) is lowered in both psbS mutant lines. This heretofore-unsuspected role for PS II-S is discussed with regard to: (1) numerous prior reports indicating plasticity of the redox potential of Q(A) and (2) the basis for the contrasting regulation of quantum yields of PS I and II in npq4-1 and npq4-9.


Injectable bone applied for ridge augmentation and dental implant placement: human progress study.
The aim of this study was to clinically evaluate the success of implants placed in conjunction with a new material, tissue-engineered bone, and the stability of the regenerated bone after functional loading on a long-term basis. The tissue-engineered bone was applied to 14 cases, in which 6 patients were with partially or totally edentulous arches scheduled for sinus floor grafting and 8 patients underwent concurrent onlay plasty. This study showed that the injectable bone formation induced bone in this anatomical site in 100% of the patients. The results also indicate that it might be possible to achieve the osseointegration of simultaneous implant placements with the grafts. It may be possible that injectable bone can shorten the period of implant treatment and reduce the patient's burden and expect good long-term prognosis.


Biophysical insights into the binding capability of Cu(II) schiff base complex with BSA protein and cytotoxicity studies against SiHa.
Herein, we have explored the effects of chlorinated mononuclear Cu(II) complex upon binding with BSA protein (bovine serum albumin) and its in vitro anti-proliferative potentiality against SiHa cell. The complex was synthesized involving a Schiff base ligand having N,N,O donor centers and characterized by several spectroscopic studies. Structure, DFT studies and Hirshfeld surface (HS) analyses were identified using crystallographic computational studies. The binding interaction with BSA depicts the efficacy of the complex towards promising binding of it with BSA. Further, the complex shows a moderate cytotoxicity against SiHa cancer cell signifying its potentiality as an anti-proliferative agent for human cervix uteri carcinoma.


Ultrasonography of tubular ectasia of the rete testis with associated scrotal findings.
Tubular ectasia of the rete testis (TERT) is a rare disorder that can be seen on ultrasonography (US) during a routine examination. In this study, we aim to discuss 15 cases of TERT and concomitant scrotal pathologies. A total of 405 patients who were admitted to our department between March 2009 and December 2011 for scrotal US with different complaints such as scrotal pain, palpable mass, infertility, varicocele, or hydrocele were evaluated. Presence or absence of TERT and associated scrotal anomalies if TERT was present were considered on gray-scale US and color Doppler US. Tubular ectasia of the rete testis was detected in 15 patients (3.7 %). It was unilateral in eleven patients (73.3 %) and bilateral in four patients (26.7 %). The mean age of the patients with TERT in this study was 43.7 years (range 24-85 years). Five patients (33.3 %) with TERT presented with infertility, four of them (26.7 %) presented with palpable mass, and six of them (40%) were admitted with scrotal pain. In 14 patients (93.3 %), at least one of the following disorders associated with TERT was seen: epididymal cyst, spermatocele, hydrocele, and varicocele. Tubular ectasia of the rete testis shows characteristic appearances on gray-scale and Doppler US evaluation, and no other modality is necessary for diagnosis. TERT is usually associated with epididymal pathologies such as epididymal cyst or spermatocele located on the same side. Testicular neoplasms, intratesticular varicocele, and cystic dysplasia of the testis should be considered in the differential diagnosis of TERT.


Fatal meningitis due to multi-resistant Haemophilus influenzae type b.
We report the case of a 3-month-old boy suffering from an acute bacterial meningitis due to a multi-resistant strain of Haemophilus influenzae type b. Also presented is our current strategy of treatment and chemoprophylaxis of Haemophilus influenzae meningitis in children.


Stabilization procedures in the patient with acute spinal cord injury.
In summary, when confronted with the spinal-cord-injured patient, the surgeon must do an initial evaluation with a complete history and physical examination, order the appropriate radiographic studies, determine the stability of the spinal column, and initiate treatment with stabilization techniques. An overview of surgical stabilization procedures used for the treatment of acute spinal cord injuries has been presented. The concept of spinal stability and the determination of the location of spinal cord compression are used to determine which surgical technique should be used. With appropriate decompression and/or stabilization procedures or both, spinal-cord-injured patients may regain additional function and begin an early rehabilitation program.


Mosquito biolarvicide production by sequential fermentation with dual strains of Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus using sewage sludge.
This study demonstrated the bioconversion of sewage sludge into a composite biolarvicide for mosquito control based on sequential fermentation with dual strains of Bacillus thuringiensis subsp. israelensis (Bti) and Bacillus sphaericus (Bs). Results showed that sewage sludge was a suitable fermentation substrate for supporting growth, sporulation and mosquitocidal proteins synthesis by Bti and Bs. Through sequential fermentation with dual strains, a 10-L bench-scale fermentor was capable of producing Bti and Bs at a cell concentration of 2.1×10(9) and 6.8×10(8) CFU/mL, respectively. Such sequential fermentation can save half of raw materials and energy consumption comparing with the sludge fermentation with single strain. The toxic activity and persistence of the composite biolarvicide against mosquito larvae in the polluted waters were enhanced by the increased toxin complexity and synergistic interactions. This study, for the first time, validates the technical feasibility of using sewage sludge to produce a cost-effective composite biolarvicide based on Bti and Bs.


Influence of physical activity on addiction parameters of rats exposed to amphetamine which were previously supplemented with hydrogenated vegetable fat.
Amphetamine (AMPH) and its derivatives are addictive drugs used to promote and enhance alertness, motivation, willingness, courage and wellbeing. However, their chronic use is related to memory loss, emotional instability, insomnia, psychosis and paranoia. In the last decades, modern society has included processed foods, rich in trans fatty acids (TFA), in their diet, what has been related to several health problems including increased AMPH preference and self-administration. In this scenario, physical activity appears to be useful to attenuate rewarding symptoms related to addictive drugs mainly by affecting brain neuroplasticity and neurotransmission. The current study has been developed to assess the influence of physical activity on addiction parameters of rats exposed to AMPH which were previously supplemented with hydrogenated vegetable fat (HVF), rich in TFA. After six weeks of HVF or soybean oil (SO, control group) supplementation, adult rats were conditioned with d,l-AMPH or vehicle for 14 days. Then, half of each experimental group was submitted to physical activity in treadmill running sessions (60min/day, 5 days/week) for 5 weeks. Animals were re-conditioned with AMPH or vehicle for 3 more days, to observe drug relapse. Locomotor activity and anxiety-like symptoms were observed 24h after the last AMPH reconditioning, and fatty acids composition was quantified in the ventral tegmental area, striatum and prefrontal cortex. All animals showed AMPH preference, but only SO sedentary showed drug relapse. No differences were observed in locomotor activity among groups, while HVF-supplemented group showed decreased exploration per se, and physical activity prevented this. Moreover, AMPH-HVF group showed increased anxiety-like symptoms, which were prevented by physical activity. These results indicate that HVF supplementation modifies AMPH addiction, whereas regular physical activity could be protective against both AMPH and TFA damages.


Comparison of the inhibitory effects of retinoids on 12-O-tetradecanoylphorbol-13-acetate-induced epidermal ornithine decarboxylase activities in CD-1 and Sencar mice.
Linear regression analysis of the dose levels of a series of retinoids required ornithine decarboxylase (ODC) induction by 50% indicated that there was no significant difference in inhibitory activity between CD-1 and Sencar mice (r = 0.9699; P less than 0.001). The ID50 values (nmol) found were as follows: retinoic acid (0.20, CD-1; 0.29, Sencar); 13-cis-retinoic acid (1.4, CD-1; 1.9, Sencar); 4-[2-(5,6,7,8-tetrahydro-5,5,8, 8-tetramethyl-2-naphthalenyl)-1E-propenyl]benzoic acid (0.04, CD-1; 0.03; Sencar); 2-[1-(4-carboxyphenyl)-1E-propeny-2-yl]-4,5,6,7-tetrahydro-4, 4-dimethylbenzothiophene (0.08, CD-1; 0.14, Sencar); 2-[1-(4-carboxyphenyl)-1E-propen-2-yl]3, 4-dihydro-4,4-dimethyl-2H-1-benzothiopyran (0.56, CD-1; 0.70, Sencar); 6-(5,6,7,8-tetrahydro-5,5,8, 8-tetramethyl-2-naphthalenyl)-2-naphthalenecarboxylic acid (1.0, CD-1; 0.90, Sencar).


Calcific valvular aortic stenosis with and without mitral anular calcium.
Seventy-six consecutive patients, mean age 66 +/- 9 years (+/- standard deviation), with isolated aortic valve replacement for calcific valvular aortic stenosis (AS) were studied. Mitral anular calcium (MAC) was detected by echocardiography in 45 patients (59%). Patients with MAC were older (p less than 0.01), had greater peak systolic aortic valve gradients (p less than 0.025), lower cardiac indexes (p less than 0.025) and smaller valve areas (p less than 0.05) than patients without MAC. Thirty-three percent of patients (15 of 45) with MAC required permanent pacemaker implantation after aortic valve replacement, compared with only 10% of patients (3 of 31) without MAC (p less than 0.025). During the follow-up of 7 years (mean 3.8 +/- 1.6), 27% of patients (12 of 45) with MAC died from cardiac causes, compared to only 7% (2 of 31) without MAC (p less than 0.025). Thus, patients with AS and MAC are older, have more severe AS, more frequently undergo permanent pacemaker implantation after aortic valve replacement and more frequently die of cardiac causes than patients with AS and no MAC during follow-up.


Dissociable hippocampal and amygdalar D1-like receptor contribution to discriminated Pavlovian conditioned approach learning.
Pavlovian conditioning is an elementary form of reward-related behavioral adaptation. The mesolimbic dopamine system is widely considered to mediate critical aspects of reward-related learning. For example, initial acquisition of positively-reinforced operant behavior requires dopamine (DA) D1 receptor (D1R) activation in the basolateral amygdala (BLA), central nucleus of the amygdala (CeA), and the ventral subiculum (vSUB). However, the role of D1R activation in these areas on appetitive, non-drug-related, Pavlovian learning is not currently known. In separate experiments, microinfusions of the D1-like receptor antagonist SCH-23390 (3.0 nmol/0.5 μL per side) into the amygdala and subiculum preceded discriminated Pavlovian conditioned approach (dPCA) training sessions. D1-like antagonism in all three structures impaired the acquisition of discriminated approach, but had no effect on performance after conditioning was asymptotic. Moreover, dissociable effects of D1-like antagonism in the three structures on components of discriminated responding were obtained. Lastly, the lack of latent inhibition in drug-treated groups may elucidate the role of D1-like in reward-related Pavlovian conditioning. The present data suggest a role for the D1 receptors in the amygdala and hippocampus in learning the significance of conditional stimuli, but not in the expression of conditional responses.


Replacement of Oct4 by Tet1 during iPSC induction reveals an important role of DNA methylation and hydroxymethylation in reprogramming.
DNA methylation and demethylation have been proposed to play an important role in somatic cell reprogramming. Here, we demonstrate that the DNA hydroxylase Tet1 facilitates pluripotent stem cell induction by promoting Oct4 demethylation and reactivation. Moreover, Tet1 (T) can replace Oct4 and initiate somatic cell reprogramming in conjunction with Sox2 (S), Klf4 (K), and c-Myc (M). We established an efficient TSKM secondary reprogramming system and used it to characterize the dynamic profiles of 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), and gene expression during reprogramming. Our analysis revealed that both 5mC and 5hmC modifications increased at an intermediate stage of the process, correlating with a transition in the transcriptional profile. We also found that 5hmC enrichment is involved in the demethylation and reactivation of genes and regulatory regions that are important for pluripotency. Our data indicate that changes in DNA methylation and hydroxymethylation play important roles in genome-wide epigenetic remodeling during reprogramming.


Relation between the tritium in continuous atmospheric release and the tritium contents of fruits and tubers.
Concentrations of organically bound tritium (OBT) and tissue-free water tritium (TFWT, also referred to as HTO) in fruits and tubers were measured at a garden plot in the vicinity of the source of chronic airborne tritium emissions during the 2008, 2010, and 2011 growing seasons. A continuous record of HTO concentration in the air moisture was reconstructed from the continuous record of Ar-41 ambient gamma radiation, as well as from frequent measurements of air HTO by active samplers at the garden plot and Ar-41 and air HTO monitoring data from the same sector. Performed measurements were used for testing the modified Specific Activity (SA) model based on the assumption that the average air HTO during the pod-filling period provides an appropriate basis for estimating the levels of OBT present in pods, fruits and tubers. It is established that the relationship between the OBT of fruits and tubers and the average air HTO from a 15-20 day wide window centred at the peak of the pod-filling period is consistent throughout the three analysed years, and could be expressed by the fruit or tuber's OBT to air-HTO ratio of 0.93 ± 0.21. For all three years, the concentration of HTO in fruits and tubers was found to be related to levels of HTO in the air, as averaged within a 3-day pre-harvest window. The variability in the ratio of plant HTO to air HTO appears to be three times greater than that for the OBT of the fruits and tubers. It is concluded that the OBT of fruits and tubers adequately follows an empirical relationship based on the average level of air HTO from the pod-filling window, and therefore is clearly in line with the modified SA approach.


[In vitro modeling of cell-scaffold interaction].
The simple approach for modeling of surface ligand - cell receptor interaction is proposed to control the effectiveness of peptide acceptor selected to be immobilized on a scaffold surface in order to promote specific cell adhesion and their subsequent proliferation and bone tissue formation. For experimental realization of such approach the affinity chromatography with use of macroporous monolithic sorbent is suggested. The biospecific GRGDSP-peptide performed the role of scaffold surface ligand which is responsible for cell adhesion, while the "cells" were simulated by polymer (polystyrene) micro particles with EDYPVDIYYLM-DLSYSMKDD-peptide immobilized on their surface. The latter peptide is the integrin molecule active site which is responsible for RGD-sequence binding. Thus the ultra-short monolithic chromatography columns (CIM-disks) represent the simplified model of a scaffold possessing biospecific properties. The qualitative evaluation of complement interaction parameters was performed via frontal analysis method followed by adsorption isotherm plotting and subsequent linearization and mathematical treatment. The data obtained reliably indicate the highly specific character of biological pair binding. This was in a good accordance with results obtained in the cell culture experiments.


Liver transplantation in a subject with familial hypercholesterolemia carrying the homozygous p.W577R LDL-receptor gene mutation.
Mutations within the low density lipoprotein (LDL)-receptor gene result in familial hypercholesterolemia, an autosomal dominant inherited disease. Clinical homozygous affected subjects die of premature coronary artery disease as early as in early childhood. We identified a girl at the age of five yr with clinical homozygous familial hypercholesterolemia presenting with achilles tendon xanthomas and arcus lipoides. Her total cholesterol reached up to 1050 mg/dL. Molecular characterization of the LDL-receptor gene revealed a homozygous p.W577R mutation. Despite intensive treatment interventions with the combination of diet, statins, colestipol, and LDL-apheresis, the patient developed symptomatic coronary artery disease at the age of 16 yr. Subsequently, orthotopic liver transplantation was performed to cure the defective LDL-receptor gene. Clinical follow-up for almost nine yr post-transplantation revealed excellent liver function, normal liver enzymes, normal LDL-cholesterol, and regression of both tendon xanthomas and symptomatic coronary artery disease. In conclusion, liver transplantation can effectively reduce LDL-cholesterol in a familial hypercholesterolemia recipient with subsequent regression of xanthomas and atherosclerosis. Timing is extremely important in these exceptional cases to exclude the demand for heart transplantation due to severe coronary artery disease. In addition, the identification of the LDL-receptor as etiology of clinical homozygous hypercholesterolemia is a prerequisite once liver transplantation is considered as therapeutic option.


Immunogenicity and toxicity of transferrin receptor-targeted hybrid peptide as a potent anticancer agent.
Transferrin receptor (TfR) is a cell membrane-associated glycoprotein involved in the cellular uptake of iron and the regulation of cell growth. Recent studies have shown elevated expression levels of TfR on cancer cells compared with normal cells. We previously designed a TfR-lytic hybrid peptide, which combines the TfR-binding peptide and a lytic peptide, and reported that it bound specifically to TfR and selectively killed cancer cells. Furthermore, the intravenous administration of TfR-lytic peptide in an athymic mouse model significantly inhibited tumor progression. To evaluate the immunogenicity of this peptide as a novel and potent anticancer agent, we investigated whether TfR-lytic hybrid peptide elicits cellular and humoral immune responses to produce antibodies. We also examined the toxicity of this peptide in syngeneic mice. We performed hematologic and blood chemistry test and histological analysis and assessed hemolytic activity to check toxicity. To evaluate the immunogenicity, measurement of murine interferon-gamma and detection of TfR-lytic-specific antibody by ELISA were demonstrated. No T cell immune response or antibodies were detected in the group treated with TfR-lytic hybrid peptide. No hematologic toxicity, except for a decrease in leukocytes, was observed, and no remarkable influence on metabolic parameters and organs (liver, kidney, and spleen) was noted. Therefore, TfR-lytic hybrid peptide might provide an alternative therapeutic option for patients with cancer.


Prediction of Potential Drug Targets and Vaccine Candidates Against Antibiotic-Resistant Pseudomonas aeruginosa.
Pseudomonas aeruginosa is one of the leading causes of nosocomial infections, characterized by increasing antibiotic resistance, severity and mortality. Therefore, numerous efforts have been made nowadays to identify new therapeutic targets. This study aimed to find potential drug targets and vaccine candidates in drug-resistant strains of P. aeruginosa. Extensive antibiotic-resistant and carbapenem-resistant strains of P. aeruginosa with complete genome were selected and ten common hypothetical proteins (HPs) containing more than 200 amino acids were obtained. The structural, functional and immunological predictions of these HPs were performed with the utility of bioinformatics approaches. Two common HPs (Gene ID: 2877781645 and 2877781936) among other investigated proteins were revealed as potential candidates for pharmaceutical and vaccine purposes based on structural and physicochemical properties, functional domains, subcellular localizations, signal peptides, toxicity, virulence factor, antigenicity, allergenicity and immunoinformatic predictions. The consequence of this predictive study will assist in novel drug and vaccine design through experimental investigations.


The prevalence of rat allergen in inner-city homes and its relationship to sensitization and asthma morbidity.
Rat allergen has proved to be an important cause of IgE-mediated hypersensitivity in the occupational setting. The prevalence and significance of rat allergen in homes has not been studied. The purpose of this study was to determine the prevalence of rat allergen in the homes of inner-city children with asthma and to examine the relationship between rat allergen exposure, sensitization, and asthma morbidity. We developed a new monoclonal-based ELISA to determine the prevalence of rat allergen in dust samples from inner-city homes of the National Cooperative Inner-City Asthma Study population. Home characteristics were evaluated to detect variables that were associated with the presence of rat allergen. Data were also analyzed to assess the relationship between the presence of rat allergen, sensitization, and asthma morbidity. Thirty-three percent of inner-city homes had detectable rat allergen (Rat n 1). The presence of rat allergen was associated with reported rat and mouse infestation, as well as evidence of mouse infestation on home inspection. Twenty-one percent of the participants were sensitized to rat allergen; however, sensitization was not more common when rat allergen was found in the home. The number of hospitalizations, unscheduled medical visits, and days with slowed activity because of asthma were significantly increased in those individuals who were both sensitized and exposed to rat allergen. Rat allergen sensitization and exposure are associated with increased asthma morbidity in inner-city children.


Desiccation tolerance: a simple process?
Water is essential for life, and thus the removal of water from a cell is a severe, often lethal stress. This is not a remarkable observation but it is one that is often taken for granted. Desiccation-tolerant cells implement structural, physiological and molecular mechanisms to survive severe water deficit. These mechanisms, and the components and pathways which facilitate them, are poorly understood. Here, recent developments are considered to illustrate the importance of desiccation, longevity and cell stasis in basic microbiology, and the relevance of the topic to the metabolic engineering of sensitive cells, including those of humans.


Control of cell division in hepatoma cells by exogenous heparan sulfate proteoglycan.
The effects of cell surface heparan sulfate proteoglycan (HSPG) prepared from log and confluent monolayers of a rat hepatoma cell line on hepatoma cell growth were studied. When HSPG isolated from confluent cells was added exogenously to log phase cells, it was internalized and free heparan sulfate (HS) chains appeared transiently in the nucleus. Concurrently, the growth of the treated cells was inhibited, but the cells resumed logarithmic growth as the level of nuclear HS fell, and the cells grew to confluence and became contact inhibited. When HSPG prepared from log-phase hepatoma cells was added exogenously to log phase cells, it was internalized but very little of the internalized HS appeared in the nucleus, and there was no change in the rate of cell growth. However, when the rate of cell growth was reduced by culture of the cells in serum- and insulin-deficient medium, HSPG prepared from log-phase cells stimulated the growth rate of these slow-growing cells. The cell cycle dependency of HSPG uptake and growth inhibition was studied in cultures synchronized by a thymidine/aphidicolin double block. When [35SO4]HSPG from confluent cells was added to synchronized cells just as they were released from the second block, a portion of the [35SO4]HSPG was internalized and [35SO4]HS appeared in the nucleus. However, at mitosis the [35SO4]HS disappeared almost completely from all of the cellular pools, and after mitosis, more of the [35SO4]HSPG was taken up and [35SO4]HS reappeared in the nucleus and remained in the nucleus until the cells divided again. When cultures were released from the aphidicolin block, both control and HSPG-treated cells progressed through the S, the G2, and the M phases of the cell cycle. However, the length of the G1 phase of the cycle was increased in the HSPG-treated cells. The treated cultures then progressed through the second S, G2, and M phases. Thus, the inhibition of cell division occurred in the G1 phase of the cell cycle, prior to the G1/S boundary. Addition of the HSPG to the synchronized cultures just after the first mitosis resulted in an immediate arrest of the cell cycle in G1.(ABSTRACT TRUNCATED AT 400 WORDS)


Psychological changes in children after anaesthesia: a comparison between halothane and ketamine.
The psychological reactions to hospitalization, anaesthesia, and operation in a group of 107 children from 1 to 12 years old, anaesthetized with ketamine or halothane after randomization were investigated through questionnaires, which the parents answered 1 month postoperatively. The percentage of replies was 96.3%. Fifty-three children were anaesthetized with ketamine and 50 with halothane. Thirteen children in the ketamine group and nine in the halothane group reacted with negative personality changes; the reactions were of less than 1 month's duration and were most frequent in the youngest children. The parents' preparation of the children had no influence on the results. The number of personality changes caused by the two anaesthetic agents did not differ significantly. Furthermore, the investigation showed the nine children reacted for the better. Thirty-six per cent of the parents felt insufficiently informed of what the hospitalization implied for their child.


Alteration of classical and hematopoiesis specific p53 pathway in the bone marrow hematopoietic stem/progenitor compartment facilitates leukemia progression in experimental mice.
Downregulation of p53 is associated with most of the neoplasms, however it claims additional significance for hematopoietic malignancy due to its supplementary role during hematopoiesis. Apart from the classical role as tumor suppressor, p53 during steady state hematopoiesis is associated with the maintenance of quiescent cell population in bone marrow by upregulating necdin (Ndn) and Gfi-1. We felt, it is necessary to delineate its attribution towards malignant conversion of hematopoietic system during leukemogenesis from all the possible angles. The present study deals with the characterization of N-N' Ethylnitrosourea (ENU) induced mouse model of leukemia by peripheral blood hemogram, bone marrow cytology, histology, cytochemical staining (MPO) and scanning electron microscopic study. We further investigated the alteration of conventional and hematopoiesis specific p53 pathways by flowcytometric expressional analysis of ATM, Chk-2, p53, p21, Ndn, Gfi-1 and Tie-2. The disruption of classical p53 pathway was observed in leukemic hematopoietic stem/progenitor population which involved downregulation of ATM, Chk-2, p53 and p21. Moreover, the expressional decline of Ndn and Gfi-1 hinted towards the mechanism of hindrance of hematopoietic quiescency in leukemic bone marrow. Increased expression of Tie-2 due to reverse correlation with p53 was found to be responsible for pathological angiogenesis in bone marrow together with increased blast burden in bone marrow during leukemia. The study presents the mechanistic scenario of the alteration of both classical as well as hematopoiesis specific p53 pathways in HSPC compartment triggering leukemic pathophysiology.


Prognostic impact of nomogram based on whole tumour size, tumour disappearance ratio on CT and SUVmax on PET in lung adenocarcinoma.
Lung adenocarcinoma frequently manifests as subsolid nodules, and the solid portion and ground-glass-opacity (GGO) portion on CT have different prognostic significance. Therefore, current T descriptor, defined as the whole tumour diameter without discrimination between solid and GGO, is insufficient. We aimed to determine the prognostic significance of solid tumour size and attempt to include prognostic factors such as tumour disappearance rate (TDR) on CT and SUVmax on PET/CT. Five hundred and ninety-five patients with completely resected lung adenocarcinoma were analyzed. We developed a nomogram using whole tumour size, TDR, and SUVmax. External validation was performed in another 102 patients. In patients with tumours measuring ≤2 cm and >2 to 3 cm, disease free survival (DFS) was significantly associated with solid tumour size (P < 0.001), but not with whole tumour size (P = 0.052). Developed nomogram was significantly superior to the conventional T stage (area under the curve of survival ROC; P = 0.013 by net reclassification improvement) in stratification of patient survival. In the external validation group, significant difference was noted in DFS according to proposed T stage (P = 0.009). Nomogram-based T descriptors provide better prediction of survival and assessment of individual risks than conventional T descriptors. • Current measurement of whole tumour diameter including ground-glass opacity is insufficient • TDR enables differentiation between invasive solid portion and non-invasive GGO portion • SUVmax demonstrates the biological aggressiveness of the tumour • We developed a nomogram using whole tumour size, TDR, and SUVmax • Nomogram-based clinical T descriptors provide better prediction of survival.


Studies on the biological activity of the dermal regeneration template.
In mammals, the skin normally responds to injury by the processes of wound contraction and scar formation. However, if a unique acellular dermal regeneration template is placed into the wound, wound contraction is strongly inhibited, and the skin undergoes a regenerative-type healing. It is proposed that these phenomena result from the disruption of a "mechanically coherent" organization of myofibroblasts within the wound. Evidence suggests that the dermal regeneration template prevents the formation of cell-extracellular matrix interactions (fibronexi), which foster the assemblage of a mechanically coherent wound bed. Additional studies support a similar mechanism for the enhanced regenerative-like response to peripheral nerves when extracellular matrix tubes are used to facilitate healing.


ALA-induced photodynamic effect on vitality, apoptosis, and secretion of vascular endothelial growth factor (VEGF) by colon cancer cells in normoxic environment in vitro.
Cancer therapy is often based on combination of conventional methods of cancer treatment with immunotherapy. Photodynamic therapy (PDT) is one of the immunomodulating methods used in oncology. We examined how PDT influences the secretory activity of colon cancer cells in vitro, especially the secretion of vascular endothelial growth factor (VEGF) in aerobic conditions. We used two cancer cell lines with different malignancy potentials: a metastatic SW620 line and a non-metastatic SW480 line. In the first stage of the experiment, we exposed each cell line to three different concentrations of photosensitizer's precursor: 5-aminolevulinic acid (ALA) and varying levels of light radiation, after which we assessed cell viability and apoptosis induction in these lines, using the MTT and LDH assays. Then, we determined the secretion of VEGF by these cells in aerobic conditions and under the ALA-PDT parameters at which cells presented the highest viability. Photodynamic treatment with ALA did not influence on VEGF secretion by the non-metastatic SW480 cells, but caused a decrease in VEGF secretion by the metastatic SW 620 cell line by 29% (p<0.05). SW 620 cell line secreted more actively VEGF than the SW480 cells, both before and after photo dynamic therapy (p<0.05). The outcome of this in vitro study presented a beneficial effect of ALA-PDT, resulting in a decrease of VEGF secretion in the more malignant SW620 cell lines. Further studies should be considered to confirm the clinical relevance of this finding.


Terraced and 3D Pyramid-Shaped Polymer Single Crystal via Low Temperature-Assisted Microfluidic Technology.
3D pyramidal polymer single crystals provide spatial gradient variations within the crystal molecules, and these variations facilitate the study of the relationship between structure and properties within the molecules of various complexes with anisotropic structures. As described herein, a low-temperature-assisted microfluidic pore channeling approach is proposed to prepare structurally ordered polymer single crystals. A mixture of dichloromethane and dimethyl sulfoxide is used as a prepolymer, and a liquid microfluidic technique is employed to grow the end-functionalized polymers into 3D polymer single crystals. Through the ordered growth of single crystals, a personalized pyramidal pattern with a homogeneous structure is formed. To evaluate the mesh node density, low-temperature growth time and substrate type are also investigated. Rectangular, pyramidal, and dendritic patterns are synthesized via low-temperature single crystal growth. This work shows that low temperature-assisted microfluidics provides a novel means to tune the 3D structure of polymer single crystals.


Chronic myocardial and coronary arterial effects of intracoronary supersaturated oxygen therapy in swine with normal and ischemic-reperfused myocardium.
The study assessed chronic myocardial, coronary and systemic effects of intracoronary supersaturated oxygen (SSO2) therapy. Left anterior descending coronary arteries of 40 swine were stented and randomized to 90-min selective intracoronary infusion of SSO2 (pO2 760-1000 mmHg) or normoxemic saline. In 20 out of 40 animals, SSO2 delivery followed a 60-min balloon occlusion to induce myocardial infarction (MI). In both normal and MI models, intracoronary treatment with hyperoxemic SSO2 therapy showed no evidence of coronary thrombosis. There were no biologically relevant differences between treatments at either time point in regard to coronary intervention site healing and neointimal growth. No signs of any myocardial or systemic toxicity were observed after 7 or 30 days. A trend was observed toward reduced incidence of microscopic MI scars and reduced infarct size in histopathology, as well as toward better recovery of echocardiographically evaluated global and regional contractility at 30 days. No treatment related infarcts or thromboemboli were observed in the downstream organs.


A case of facial hemiatrophy associated with linear scleroderma and homolateral masseter spasm.
The rare combination of progressive facial hemiatrophy, linear scleroderma and spasm of the homolateral masseter muscle was observed in a 39 year old woman. The possible pathogenetic meaning of the association is discussed on the electrophysiological evidence and in the light of published cases.


Regulation of Tissue-Specific Alternative Splicing: C. elegans as a Model System.
Alternative pre-mRNA splicing serves as an elegant mechanism for generating transcriptomic and proteomic diversity between cell and tissue types. In this chapter, we highlight key concepts and outstanding goals in studies of tissue and cell-specific alternative splicing. We place particular emphasis on the use of C. elegans as a tractable model organism for in vivo studies of alternative splicing between tissues and also at single cell resolution. We describe our current understanding of tissue and cell-specific regulation in the animal, and emerging techniques that will allow for future mechanistic studies as well as systems level investigations of spatio-temporal splicing under laboratory conditions and in response to environmental stimuli.


Effects of latitude, host body size, and host trophic guild on patterns of diversity of helminths associated with humans, wild and domestic mammals of Mexico.
Parasites are strictly associated with their hosts and present a great diversity of life histories, often resulting in different diversity patterns than those observed in free-living species. However, ecological approaches have detected that, in some cases, mammal-associated helminths respond similarly to non-parasitic species in terms of diversity patterns. Using 2200 recorded interactions, we analysed the diversity patterns of helminths (Acanthocephala, Nematoda and Platyhelminthes) harbored by humans, wild and domestic mammals of Mexico, depending on latitude, host body mass and trophic guild (carnivore, herbivore, insectivore, omnivore), considering helminth richness and average taxonomic distinctness, and host phylogenetic diversity and phylogenetic clustering. Latitude was positively correlated with the average taxonomic distinctness encompassing the three parasite phyla and nematodes. Northern latitudes had less taxonomically related parasite assemblages. Host body mass had a significant negative relationship with the average taxonomic distinctness of acanthocephalans and the richness of helminths associated to wild hosts. The omnivore hosts had greater parasite richness, while insectivores had a less taxonomically related parasite assemblage and herbivores had a more heterogeneous parasite assemblage. Our results highlight the importance of incorporating different dimensions of diversity, such as average taxonomic distinctness and to consider the composition of parasite assemblages to better understand their diversity patterns.


Prognostication in Stargardt Disease Using Fundus Autofluorescence: Improving Patient Care.
To explore fundus autofluorescence (FAF) imaging as an alternative to electroretinography as a noninvasive, quick, and readily interpretable method to predict disease progression in Stargardt disease (STGD). Retrospective case series of patients who attended Moorfields Eye Hospital (London, United Kingdom). Patients with STGD who met the following criteria were included: (1) biallelic disease-causing variants in ABCA4, (2) electroretinography testing performed in house with an unequivocal electroretinography group classification, and (3) ultrawidefield (UWF) FAF imaging performed up to 2 years before or after the electroretinography. Patients were divided into 3 electroretinography groups based on retinal function and 3 FAF groups according to the extent of hypoautofluorescence and retinal background appearance. Fundus autofluorescence images of 30° and 55° were reviewed subsequently. Electroretinography and FAF concordance and its association with baseline visual acuity (VA) and genetics. Two hundred thirty-four patients were included in the cohort. One hundred seventy patients (73%) were in electroretinography and FAF groups of the same severity, 33 (14%) were in a milder FAF than electroretinography group, and 31 (13%) were in a more severe FAF than electroretinography group. Children < 10 years of age (n = 23) showed the lowest electroretinography and FAF concordance at 57% (9 of the 10 with discordant electroretinography and FAF showed milder FAF than electroretinography), and adults with adult onset showed the highest (80%). In 97% and 98% of patients, 30° and 55° FAF imaging, respectively, matched with the group defined by UWF FAF. We demonstrated that FAF imaging is an effective method to determine the extent of retinal involvement and thereby inform prognostication by comparing FAF with the current gold standard of electroretinography. In 80% of patients in our large molecularly proven cohort, we were able to predict if the disease was confined to the macula or also affected the peripheral retina. Children assessed at a young age, with at least 1 null variant, early disease onset, poor initial VA, or a combination thereof may have wider retinal involvement than predicted by FAF alone, may progress to a more severe FAF phenotype over time, or both. Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.


Mechanism of Assembly of a Substrate Transfer Complex during Tail-anchored Protein Targeting.
Tail-anchored (TA) proteins, defined as having a single transmembrane helix at their C terminus, are post-translationally targeted to the endoplasmic reticulum membrane by the guided entry of TA proteins (GET) pathway. In yeast, the handover of TA substrates is mediated by the heterotetrameric Get4/Get5 complex (Get4/5), which tethers the co-chaperone Sgt2 to the targeting factor, the Get3 ATPase. Binding of Get4/5 to Get3 is critical for efficient TA targeting; however, questions remain about the formation of the Get3·Get4/5 complex. Here we report crystal structures of a Get3·Get4/5 complex from Saccharomyces cerevisiae at 2.8 and 6.0 Å that reveal a novel interface between Get3 and Get4 dominated by electrostatic interactions. Kinetic and mutational analyses strongly suggest that these structures represent an on-pathway intermediate that rapidly assembles and then rearranges to the final Get3·Get4/5 complex. Furthermore, we provide evidence that the Get3·Get4/5 complex is dominated by a single Get4/5 heterotetramer bound to one monomer of a Get3 dimer, uncovering an intriguing asymmetry in the Get4/5 heterotetramer upon Get3 binding. Ultrafast diffusion-limited electrostatically driven Get3·Get4/5 association enables Get4/5 to rapidly sample and capture Get3 at different stages of the GET pathway.


Biological aspects of Lyme disease spirochetes: unique bacteria of the Borrelia burgdorferi species group.
Borrelia burgdorferi sensu lato is a group of at least twelve closely related species some of which are responsible for Lyme disease, the most frequent zoonosis in Europe and the USA. Many of the biological features of Borrelia are unique in prokaryotes and very interesting not only from the medical viewpoint but also from the view of molecular biology. Relevant recent articles were searched using PubMed and Google search tools. This is a review of the biological, genetic and physiological features of the spirochete species group, Borrelia burgdorferi sensu lato. In spite of a lot of recent articles focused on B. burgdorferi sensu lato, many features of Borrelia biology remain obscure. It is one of the main reasons for persisting problems with prevention, diagnosis and therapy of Lyme disease. The aim of the review is to summarize ongoing current knowledge into a lucid and comprehensible form.


Electrophysiological study of synaptic connections between a transplanted lateral geniculate nucleus and the visual cortex of the host rat.
The lateral geniculate nucleus (LGN) of a fetal rat was transplanted to the visual cortex (VC) of a neonatal rat. A current source-density analysis of field potentials and an intracellular study of neuronal responses were conducted in slice preparations by electrical stimulation of transplanted LGN and host VC. The results indicated that synaptic connections were established reciprocally between the transplanted LGN and the host VC.


Pregnancy and Kidney Diseases: Multidisciplinary Follow-Up and the Vicious Circles Involving Pregnancy and CKD, Preeclampsia, Preterm Delivery and the Kidneys.
Thomas Addis, the father of nephrology, once wrote that a clinician is complex, "he is part craftsman, part practical scientist, and part historian" [...].


A new enzyme immunoassay of microsomal rat liver epoxide hydrolase.
Antiserum against purified rat liver microsomal epoxide hydrolase was produced in the rabbit. We developed an enzyme-linked immunosorbent assay which is reliable with regard to its analytical criteria. The concentration of epoxide hydrolase was measured in liver microsomes of control rats and animals treated with F 1379 (250 mg/kg/day) for 5, 7, 14, and 21 days. This hypolipidemic drug was able to induce strong epoxide hydrolase activity and enhance protein concentration. The gradual increase in epoxide hydrolase concentration paralleled the increase of epoxide hydrolase activity, with stabilization occurring after the 14th until the 21st day of treatment.


Dietary control of the chain elongation of palmityl-CoA in rat liver microsomes.
The rate of chain elongation of palmityl-CoA to stearyl-CoA in rat liver microsomes was studied in connection with the nutritional status of the rats. The microsomal chain elongation activity, which had been decreased by starvation for 48 hr, was rapidly increased to a high level on refeeding. The apparent Km value for malonyl-CoA in both normal and refed rats was the same, 1.2 X 10(4)M. Both cycloheximide and actinomycin D prevented the induction of microsomal chain elongation activity which was associated with refeeding. In addition, the activity of acyl-CoA hydrolase and the rates of esterification of acyl-CoA into phospholipids and neutral lipids in microsomes were not changed by the dietary alteration. These results support the conclusion that changes of the activity of microsomal chain elongation of palmityl-CoA in various nutritional status result from a rapid synthesis of new enzyme(s).


Hydrothermal synthesis of a 3D double-sided comb-like ZnO nanostructure and its growth mechanism analysis.
Self-assembly of two-dimensional (2D) nanosheets and one-dimensional (1D) nanorods into three-dimensional (3D) double-sided comb-like ZnO nanostructures has been successfully performed via a simple one-step hydrothermal method on Si and ITO substrates. It is found that the thickness of the ZnO seeds plays a vital role in the formation of 3D double-sided comb-like ZnO.


Influence of the activity of glutathione reductase on the response of cultured lens epithelial cells from young and old rabbits to hydrogen peroxide.
Our previous studies on cultured rabbit lens epithelial cells from 4-day-old rabbits showed that the glutathione redox cycle plays an important role in detoxifying H2O2, a potentially damaging oxidant present in the aqueous humor. Here we report the effect of donor age and cell density on the ability of cultured rabbit lens epithelial cells to detoxify H2O2. Lens epithelial cells (8 x 10(5] from a 4-day-old and an 8-year-old rabbit were cultured for 3 hr in minimal essential medium (MEM) or in MEM containing 0.01-0.1 mM H2O2 maintained with glucose oxidase. We determined the effect of H2O2 on the level of reduced glutathione (GSH), hexose monophosphate shunt activity, cell growth, and morphology. For growth studies, cells were exposed to the desired concentration of H2O2 for 3 hr and then cultured in MEM plus 10% rabbit serum for 7 days and counted. Young and old untreated cells contained high levels (30-40 nmol/8 x 10(5) cells) of GSH. Cells from 4-day-old rabbits tolerated 0.03 mM H2O2 with no effect on GSH and a minimal decrease in subsequent cell growth. However, in the older cells, GSH and growth were substantially diminished following treatment with 0.03 mM H2O2. Cells plated out at high density (8 x 10(5] were more tolerant of 0.03 mM H2O2 than cells plated out at low density (5 x 10(4]. Maximum shunt activity in the younger cells exposed to H2O2 was twice that of the older cells and occurred at a higher level of H2O2 (0.04 compared with 0.03 mM). Enzyme activities in untreated young and old cells were comparable for hexokinase, glucose-6-phosphate dehydrogenase, and glutathione peroxidase. However, glutathione reductase activity was 50% lower in the cells from the 8-year-old rabbit. The toxicity of H2O2 to cultured lens epithelial cells was directly related to donor age and inversely related to cell density. The damage in the older lens epithelial cells at 0.03 mM H2O2 was apparently due, in part, to a diminished response of the glutathione redox cycle to oxidative challenge.


The effects of different intraocular irrigating solutions on the apoptosis of cultured RPE cells.
To investigate the effect on porcine retinal pigment epithelial (RPE) cells of modified intraocular irrigating solutions compared with BSS and BSS plus. Confluent cultures of RPE cells were incubated in experimental intraocular irrigating solutions. The cells were then examined for breakdown of DNA by the TUNEL procedure. Fragmentation of the DNA from cells was also confirmed by flow cytometry. Most of the experimentally, treated cells exhibited a shrunken appearance for up to 72 h. There was a steady increase in the number of cells labeled by the TUNEL method in three solutions with time. The data demonstrated that the influence of solutions on the percentage of RPE cell nuclei that gave a clear positive TUNEL stain was, in ascending order: modified solution>BSS>BSS plus. Of the three solutions tested, BSS plus showed least apoptosis. It is suggested that BSS plus is less harmful to RPE cells than the other solutions tested. Introduction of other adjunctive solutions of antibiotics, mydriatics, miotics, and steroid, resulting in an altered electrolyte balance, pH, or osmolality in the solution, may compromise the safety and efficacy of a properly formulated and packaged solution.


Clinicopathologic differences between carcinoma in the gastric remnant stump after distal partial gastrectomy for benign gastroduodenal lesions and primary carcinoma in the upper third of the stomach.
This study evaluated the prognosis of patients with carcinoma in the gastric remnant stump after distal gastrectomy for benign disease (stump cancer). Twenty patients with resected stump cancer were studied and compared with 266 patients with resected primary cancer in the upper third of the stomach (primary cancer). The percentages of lymph nodes with metastases located at the lesser curvature and the left gastric artery, respectively, were 50.0% and 25.2% in patients with primary cancer and 15.0% and 5.0% in patients with stump cancer. In patients with remnant gastric cancer, lymph node metastases at jejunal mesentery near anastomotic site were found in 2 of 20 (10.0%). The 5-year survival rates were 62.1% and 52.5% for patients with curatively resected primary cancer (n = 203) and stump cancer (n = 17), respectively, and there was no statistical difference between the two groups. There was a distinct difference in terms of patterns of recurrence between patients with curatively resected primary cancer and those with stump cancer. The percentages of peritoneal and hematogenic metastases, respectively, were 40.6% and 31.3% in primary cancer and 0% and 83.3% in stump cancers. The lymph flow from the remnant stomach seems to be changed by the initial operation, and as a result, in stump cancer, metastatic lymph nodes are located at different places from those associated with primary cancer. Even when curative surgery is performed in patients with stump cancer, it is important to follow up patients and check for possible metastasis to the liver.


Storing, retrieving, and processing optical information by Raman backscattering in plasmas.
By employing stimulated Raman backscattering in a plasma, information carried by a laser pulse can be captured in the form of a very slowly propagating plasma wave that persists for a time long compared with the pulse duration. If the plasma is then probed with a short laser pulse, the information stored in the plasma wave can be retrieved in a second scattered electromagnetic wave. The recording and retrieving processes can conserve robustly the pulse shape, thus enabling the recording and retrieving with fidelity of information stored in optical signals.


Overexpression of integrin alpha6 and beta4 enhances adhesion and proliferation of human retinal pigment epithelial cells on layers of porcine Bruch's membrane.
Transplantation of retinal pigment epithelium (RPE) following removal of choroidal neovascular membranes has been attempted in patients with age-related macular degeneration (AMD). However, inability of transplanted RPE to initially attach and subsequently proliferate on Bruch's membrane may lead to failure of RPE transplants and poor visual outcomes. Integrin alpha(6)beta(4) functions as a receptor for laminin, the major component of Bruch's membrane, and mediates the stable attachment of most epithelial cells to the underlying basement membrane. To improve adhesion and proliferation of transplanted RPE on Bruch's membrane, we elucidated the roles of integrin alpha(6)beta(4) in RPE adhesion to extracellular matrix and investigated whether ex vivo gene transfer of integrin alpha(6) and beta(4) in RPE could promote adhesion and proliferation of transplanted RPE on Bruch's membrane. The expression of integrin alpha(6) and beta(4) mRNA and surface protein in ARPE-19 cells was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometric analysis. We generated point mutation in the ligand binding domain of integrin alpha(6) and beta(4) by using site-directed mutagenesis and transfected these mutated constructs into ARPE-19 cells. Adhesion assay was used to determine the roles of integrin alpha(6) and beta(4) in RPE adhesion to extracellular matrix. In addition, we transfected full-length alpha(6) cDNA or beta(4) cDNA into ARPE-19 cells. The reattachment and proliferation ratios of alpha(6)-cDNA- or beta(4)-cDNA-transfected ARPE-19 cells on different layers of Bruch's membrane were determined by cell adhesion and proliferation assays. Cell morphology and surface coverage were evaluated by scanning electron microscopy 7 days after plating on various layers of Bruch's membrane. We found that integrin alpha(6) and beta(4) mRNA and proteins were constitutively expressed in ARPE-19 cells. Decreased endogenous integrin alpha(6) and beta(4) expression by selective mutation of amino acid residues caused a significant reduction in adhesion of ARPE-19 cells to laminin 5. Modification of integrin expression by transfection of alpha(6) cDNA into ARPE-19 cells induced a significant increase in cell adhesion to laminin 5, fibronectin, whereas transfection with beta(4) cDNA caused increased adhesion only to laminin 5. alpha(6)-cDNA-transfectants increased cell attachment and proliferation on all layers of Bruch's membrane, whereas beta(4)-cDNA-transfectants enhanced adhesion and proliferation on basal lamina and inner collagenous layers. These data indicate that integrin alpha(6) and beta(4) play a role in adhesion of ARPE-19 cells to extracellular matrix. Modification of integrin expression by ex vivo genetic manipulation in RPE might be an alternative strategy to increase the success of RPE transplantation.


[Lymphocytic and non-lymphocytic dysplasia, carcinoma in situ and infiltrating squamous cell carcinoma of the cervix (author's transl)].
The degree of lymphocytic infiltration in 490 cases of infiltrating squamous cell carcinoma, carcinoma in situ, and dysplasias of the cervix was studied. 204 cases did not show any lymphocytic infiltration at the site of dysplasias, carcinoma in situ, and infiltrating squamous cell carcinoma. The degree of lymphocytic infiltration was furthermore not related to the grade of dysplasia. Taking into account all cases it was found that 75% of carcinoma in situ and infiltrating squamous cell carcinoma remained without or with only sparse lymphocytic infiltrations.


The rising incidence of heterotopic pregnancy: two case reports.
Two cases of associated intrauterine and extrauterine pregnancy are presented. In the past combined pregnancy was thought to be a rare obstetric event, with an estimated theoretical incidence of one in 30,000 gestations. A review of the medical records of the Sint-Augustinus Hospital revealed 3 cases of combined pregnancy out of a total of 12,338 pregnancies during the 5-year period January 1981 to December 1985. These data confirm recent communications which state that literature concerning heterotopic gestations has greatly underestimated its rate of occurrence in Western countries. For patients who have been treated with ovulation-inducing medication, the theoretical risk is particularly substantial.


Lyophilization protects [FeFe]-hydrogenases against O2-induced H-cluster degradation.
Nature has developed an impressive repertoire of metal-based enzymes that perform complex chemical reactions under moderate conditions. Catalysts that produce molecular hydrogen (H2) are particularly promising for renewable energy applications. Unfortunately, natural and chemical H2-catalysts are often irreversibly degraded by molecular oxygen (O2). Here we present a straightforward procedure based on freeze-drying (lyophilization), that turns [FeFe]-hydrogenases, which are excellent H2-producers, but typically extremely O2-sensitive in solution, into enzymes that are fully resistant against O2. Complete dryness protects and conserves both, the [FeFe]-hydrogenase proteins and their inorganic active-site cofactor (H-cluster), when exposed to 100% O2 for days. The full H2-formation capacity is restored after solvation of the lyophilized enzymes. However, even minimal moisturizing re-establishes O2-sensitivity. The dry [FeFe]-hydrogenase material is superior also for advanced spectroscopic investigations on the H-cluster reaction mechanism. Our method provides a convenient way for long-term storage and impacts on potential biotechnological hydrogen production applications of hydrogenase enzymes.


Effect of metals on the adsorption and extractability of 14C-phenanthrene in soils.
The fate of polycyclic aromatic hydrocarbons (PAHs) in contaminated soils may be affected by several environmental factors including the presence of co-contaminants. This study was conducted in order to assess the effect of metals on (i) the adsorption of 14C-phenanthrene in soils and (ii) its extractability and ability to form non-extractable residues. The first objective was accomplished using batch adsorption experiments with an uncontaminated agricultural soil spiked with the metals Cd, Cu, Pb, and Zn. Adsorption of phenanthrene was significantly higher after the addition of the metals (Kf = 21.48 vs. 8.55) and the desorption less readily reversible when compared to the unspiked soil. The extractability of phenanthrene was assessed with incubation (4 months, laboratory conditions) and microlysimeter experiments (6 months, natural climatic conditions) on three soils spiked with metals. All the soils were labelled with 14C-phenanthrene. The amount of extractable phenanthrene residues was significantly higher when the metals had been added to the soils. Nevertheless, the quantity of non-extractable residues was non-significantly different between the spiked and unspiked soils. The mechanism leading to increased adsorption and extractability of phenanthrene in the presence of metals is still unknown. In perspective, it would be interesting to assess the bioavailability of PAHs in the presence of metals in further experiments.


A Gate Programmable van der Waals Metal-Ferroelectric-Semiconductor Vertical Heterojunction Memory.
Ferroelectricity, one of the keys to realize non-volatile memories owing to the remanent electric polarization, is an emerging phenomenon in the 2D limit. Yet the demonstrations of van der Waals (vdW) memories using 2D ferroelectric materials as an ingredient are very limited. Especially, gate-tunable ferroelectric vdW memristive device, which holds promises in future multi-bit data storage applications, remains challenging. Here, a gate-programmable multi-state memory is shown by vertically assembling graphite, CuInP2 S6 , and MoS2 layers into a metal(M)-ferroelectric(FE)-semiconductor(S) architecture. The resulted devices seamlessly integrate the functionality of both FE-memristor (with ON-OFF ratios exceeding 105 and long-term retention) and metal-oxide-semiconductor field effect transistor (MOS-FET). Thus, it yields a prototype of gate tunable giant electroresistance with multi-levelled ON-states in the FE-memristor in the vertical vdW assembly. First-principles calculations further reveal that such behaviors originate from the specific band alignment between the FE-S interface. Our findings pave the way for the engineering of ferroelectricity-mediated memories in future implementations of 2D nanoelectronics.


The effect of pre-maturation culture using phosphodiesterase type 3 inhibitor and insulin, transferrin and selenium on nuclear and cytoplasmic maturation of bovine oocytes.
This study aims to evaluate if a pre-maturation culture (PMC) using cilostamide as a meiotic inhibitor in combination with insulin, transferrin and selenium (ITS) for 8 or 24 h increases in vitro embryo production. To evaluate the effects of PMC on embryo development, cleavage rate, blastocyst rate, embryo size and total cell number were determined. When cilostamide (20 μM) was used in PMC for 8 or 24 h, 98% of oocytes were maintained in germinal vesicles. Although the majority of oocytes resumed meiosis after meiotic arrest, the cleavage and blastocyst rates were lower than the control (P 0.05) to the control. The deleterious effect of 20 μM cilostamide treatment for 24 h on a PMC was confirmed by lower cumulus cell viability, determined by trypan blue staining, in that group compared with the other groups. A lower concentration (10 μM) and shorter exposure time (8 h) minimized that effect but did not improve embryo production. More studies should be performed to determine the best concentration and the arresting period to increase oocyte competence and embryo development.