Attachment insecurity mediates the relationship between childhood trauma and eating disorder psychopathology in a clinical sample: a structural equation model.
Childhood maltreatment occurs often among those with an eating disorder and is considered a nonspecific risk factor. However, the mechanisms by which childhood maltreatment may lead to an eating disorder are not well understood. The current study tests a model in which attachment insecurity is hypothesized to mediate the relationship between childhood maltreatment and eating disorder psychopathology. Treatment seeking adults with eating disorders (N=308) completed questionnaires about childhood maltreatment, eating disorder psychopathology, and adult attachment. Structural equation models indicated that childhood trauma had a direct effect on eating disorder symptoms. Also, attachment anxiety and avoidance each equally mediated the childhood maltreatment to eating disorder psychopathology relationship. Attachment insecurity, characterized by affect dysregulation and interpersonal sensitivities may help to explain why eating disorder symptoms may be one consequence of childhood maltreatment in a clinical sample. Clinicians treating primarily those with trauma might assess for disordered eating as a potential manifestation of the sequelae of trauma and attachment insecurity.


Sequence and expression of Sox-18 encoding a new HMG-box transcription factor.
The newly identified Sox gene family (Sry-like HMG-box gene) is characterized by a conserved DNA sequence encoding a domain of approx. 80 amino acids (aa) which is responsible for sequence-specific DNA binding. The first member isolated, the mammalian Y-linked testis-determining gene, Sry, is necessary and sufficient for male development. We report here the identification of two new members of this family, Sox-17 and 18. We have determined the full cDNA sequence of Sox-18 which encodes a protein of 378 aa. Sox-18 mRNA transcripts were restricted to heart, lung and skeletal muscle in the adult mouse.


Rhesus and Kell Phenotyping of Voluntary Blood Donors: Foundation of a Donor Data Bank.
To assess the Rhesus (Rh) and Kell (K) phenotype of voluntary blood donors and lay foundation of a data bank of voluntary blood donors. Cross-sectional study. Blood Bank, The Aga Khan University Hospital, Karachi, in the year 2014. Voluntary blood donors were inducted after taking written informed consent. Three -5cc of EDTA anticoagulated blood sample was taken to phenotype red cells for C, c, E, e, and Kell antigens using antisera. [DiaMedSwitzerland]. Hundred blood donors were included in the study. ABO blood groups of the donors were: O [37%], B [31%], A [21%] and AB [11%]. Ninety-seven percent were Rh D positive while 3% were Rh D negative; 'e' antigen had the highest frequency [99%], while 'E' antigen was the least frequent [19%]. The most common probable Rh phenotype was R1R1 ((DCe/DCe) in 44 [44%]. In the Kell system, all the donors [100%] had phenotype of K-k+. The most common blood group was O +ve. The pattern of Rhesus antigen expression and phenotype found in this study was concordant to that reported previously from Asia. However, there was a much lower frequency of K antigen.


Localization of viral DNA-replication in sections of human warts by nucleic acid hybridization with complementary RNA of human papilloma virus Type 1.
Using complementary RNA of human papilloma virus type 1 (HPV1) and in situ hybridization techniques the localization of viral DNA replication was studied in sections of 38 human virus acanthomata from 31 different patients. In five cases significant labeling was detected by autoradiography. Labeling started always in the first or the second suprabasal cell layer and was strongly limited to the nuclei. A remarkable early beginning of the vacuolated process seemed to be correlated with the visible DNA replication. No labeling could be detected in the basal cell layer. This suggests that these cells contain at the most only a small number of viral genomes. Our findings represent only the situation in human warts definitely caused by HPV 1. We are not able to say, whether our negative hybridization results in the remaining warts indicate either the scarcity of viral genomes within these sections or their infection by a different agent.


Analysis of food packaging UV inks for chemicals with potential to migrate into food simulants.
Ultraviolet (UV) inks are an alternative formulation system to the more usual paste or liquid inks (oils or solvents based) that dry mainly by evaporation or penetration into the printed substrate. Based on acrylic acid chemistry, UV inks dry (the exact term is 'curing') by the chemical process of photopolymerization. Their composition (acrylate monomers and oligomers together with photo-initiators) exposed to UV emission lamps on the printing press units enable the transformation of the freshly printed ink layer into a tack-free film. For UV inks intendedfor primary food packaging, special care has to be paid to potential migrating species like small photo-initiator molecules and acrylate monomers not cross-linked in the formed network. The paper presents chromatographic methods to ascertain the level of ink ingredients potentially available to migrate into food simulants (migration tests). GC/MS was employed to quantify the levels of photo-initiators or acrylic esters (acrylates).


Adenovirus-directed ocular innate immunity: the role of conjunctival defensin-like chemokines (IP-10, I-TAC) and phagocytic human defensin-alpha.
Adenovirus (Ad), a major cause of viral conjunctivitis worldwide, is controlled initially by the innate immune response on the ocular surface. In the present study, cultured human conjunctival epithelial cells were used to identify host genes responsive to infection by a clinical isolate of Ad5, and the antiviral activity of some of their peptide products was investigated. Primary human conjunctival epithelial cells in culture were infected with Ad5 at high (1.0), low (0.1), or zero (0.0) multiplicities of infection (MOI) and harvested at different times after infection (1.5, 6, and 16 hours). Host gene expression was profiled with oligonucleotide microarrays (GeneChips; Affymetrix, Santa Clara, CA). Peptide products of interest were tested for antiviral activity in direct inhibition assays against respiratory serotypes (Ad3, Ad5), ocular serotypes (Ad8, Ad19), and HSV-1. At high MOI, viral infection suppressed the interferon (IFN)-mediated host antiviral response seen at low MOI. At 16 hours after infection, 63 unique characterized transcripts were identified that were robustly and significantly suppressed by high MOI, (i.e., MOI [0.1]/MOI [0.1] > or = 2, P < 0.006). Of these, 29 (46%) transcripts are involved in IFN signaling or are IFN or virus induced. This subset included CXCL10 and CXCL11, encoding IP-10 and I-TAC, respectively. These defensin-like chemokines and human alpha-defensin-1 directly inhibited Ad3 and Ad5 but not Ad8 or Ad19. In response to low-level Ad5 infection, conjunctival epithelial cells showed upregulation of IFN-associated genes. The peptide products of two of these, IP-10 and I-TAC, are directly active against Ad3, and IP-10 is active against Ad5. The ocular tropism of Ad8 and Ad19 may be due in part to their resistance to these defensin-like chemokines.


Conserved factor Dhp1/Rat1/Xrn2 triggers premature transcription termination and nucleates heterochromatin to promote gene silencing.
Cotranscriptional RNA processing and surveillance factors mediate heterochromatin formation in diverse eukaryotes. In fission yeast, RNAi machinery and RNA elimination factors including the Mtl1-Red1 core and the exosome are involved in facultative heterochromatin assembly; however, the exact mechanisms remain unclear. Here we show that RNA elimination factors cooperate with the conserved exoribonuclease Dhp1/Rat1/Xrn2, which couples pre-mRNA 3'-end processing to transcription termination, to promote premature termination and facultative heterochromatin formation at meiotic genes. We also find that Dhp1 is critical for RNAi-mediated heterochromatin assembly at retroelements and regulated gene loci and facilitates the formation of constitutive heterochromatin at centromeric and mating-type loci. Remarkably, our results reveal that Dhp1 interacts with the Clr4/Suv39h methyltransferase complex and acts directly to nucleate heterochromatin. Our work uncovers a previously unidentified role for 3'-end processing and transcription termination machinery in gene silencing through premature termination and suggests that noncanonical transcription termination by Dhp1 and RNA elimination factors is linked to heterochromatin assembly. These findings have important implications for understanding silencing mechanisms targeting genes and repeat elements in higher eukaryotes.


Grp78 is required for intestinal Kras-dependent glycolysis proliferation and adenomagenesis.
In development of colorectal cancer, mutations in APC are often followed by mutations in oncogene KRAS The latter changes cellular metabolism and is associated with the Warburg phenomenon. Glucose-regulated protein 78 (Grp78) is an important regulator of the protein-folding machinery, involved in processing and localization of transmembrane proteins. We hypothesize that targeting Grp78 in Apc and Kras (AK)-mutant intestines interferes with the metabolic phenotype imposed by Kras mutations. In mice with intestinal epithelial mutations in Apc, Kras G12D and heterozygosity for Grp78 (AK-Grp78 HET ) adenoma number and size is decreased compared with AK-Grp78 WT mice. Organoids from AK-Grp78 WT mice exhibited a glycolysis metabolism which was completely rescued by Grp78 heterozygosity. Expression and correct localization of glucose transporter GLUT1 was diminished in AK-Grp78 HET cells. GLUT1 inhibition restrained the increased growth observed in AK-mutant organoids, whereas AK-Grp78 HET organoids were unaffected. We identify Grp78 as a critical factor in Kras-mutated adenomagenesis. This can be attributed to a critical role for Grp78 in GLUT1 expression and localization, targeting glycolysis and the Warburg effect.


Nutritional values of different blood diets expressed as reproductive potentials in adult Stomoxys calcitrans L. (Diptera: Muscidae).
Experiments with Stomoxys calcitrans adults showed that different blood diets markedly affect the lifespan and reproductive potential of this species. When fed on blood from herbivores (cattle, sheep, goat, horse and donkey) the adults lay more eggs than they do when fed on blood from omnivores (pig). Blood from carnivores (dog) is even less suitable than that from omnivores, and no eggs are laid when the flies are fed on chicken blood.


Increased somatostatin mRNA expression in periventricular nucleus of rat hypothalamus during hypoxia.
We reported that hypoxia inhibited the growth hormone (GH) and induced somatostatin (SS) release from the hypothalamic median eminence (ME) of rats. This study is designed to examine the SS mRNA alterations in the periventricular nucleus (PeN) of the hypothalamus in rats and the possible involvement of glucocorticoid (GC) during hypoxia. Rats were exposed to hypoxia in a simulated hypobaric chamber. SS mRNA levels in the PeN were tested by in situ hybridization. Hypoxia of 5-km altitude (10.8% O(2)) for 2, 5 and 24 h increased the SS mRNA expression by 34.72%, 50.31% and 95.05% (p<0.05), respectively. Severe hypoxia of 7-km altitude (8.2% O(2)) enhanced the SS expression by 79.08% (p<0.01), 74.90% (p<0.01) and 71.40% (p<0.05), respectively. Prolonged hypoxia (5 km for 5 days) exposure augmented a 2.5-fold SS mRNA (p<0.001). One week post adrenalectomy (ADX), SS mRNA level was significantly increased. During hypoxia, 5 km for 5 h, SS mRNA in ADX rats was not further increased. An increased SS mRNA was showed by pretreatment with low dose of dexamethasone (DEX) (125 microg/kg, i.p.) to ADX animals but this increase was depressed by a high dose of DEX (500 microg/kg, i.p.). The data suggested that (1) hypoxia stimulated the expression of SS mRNA in the PeN of rat hypothalamus. (2) Increased circulating GC levels might play a role in upregulating the SS mRNA in the rat PeN during hypoxia.


Azorella selago Hook. used to estimate glacier fluctuations and climatic history in the Kerguelen Islands over the last two centuries.
Climatic variations over the two last centuries are well known for the northern hemisphere, but very little information is available for subantarctic islands. In this paper, we combined geomorphological observations and a new biological dating technique to propose a reconstruction of the cool and warm events in the Kerguelen Islands during the last two centuries. The usual dating methods, such as dendrochonology or C14 dating, are not applicable on Kerguelen. Therefore, the radial growth ofAzorella selago Hook., a cushion-forming Umbelliferae species, was used to estimate the absolute age of deglaciated areas. Glacial margins in the vicinity of the Glacier Ampère constitute the most complete chronosequence studied in this part of the world and illustrate seven warming-cooling cycles. This new dating technique is validated by the close relationship between the calculated ages of these climatic events and the results of several studies in other circumantarctic regions. The Glacier Ampère reached its maximum extent at the end of the eighteenth century. Since 1799, two discrete phases may be distinguished: the first period (1799-1965) is characterized by small glacier fluctuations (1 km retreat overall) whereas in the second period (1966 to the present), the retreat is much more rapid (about 3 km). It seems that the current dramatic glacial retreat on Kerguelen is related to a major change in the climate and could illustrate a more general southern hemispheric pattern of glacial fluctuations.


Porphyromonas gingivalis is a risk factor for the development of nonalcoholic fatty liver disease via ferroptosis.
Nonalcoholic fatty liver disease (NAFLD) is a metabolic liver disease that can eventually lead to liver cirrhosis and hepatocellular carcinoma. Porphyromonas gingivalis (P.g) is the main pathogen that causes periodontal disease, which participates in the development of NAFLD. The purpose of our study was to further study the direct role of P.g in NAFLD and the underlying molecular mechanism. An animal model of oral P.g administration was established, and liver function and pathology in this model were evaluated. The gut microbiome and metabolic products were analysed. Furthermore, the Th17/Treg balance in the spleen and liver was assessed. In our study, NAFLD was observed in all the mice that were orally administered P.g. The gut microbiome and metabolic products were altered after oral P.g administration. P.g and ferroptosis were observed in the livers of the mice after oral P.g administration. Additionally, ferroptosis was observed in hepatocytes in vitro, but it was reversed by ferroptosis inhibitors. In addition, P.g triggered an imbalance in the Th17/Treg ratio in the liver and spleen in vivo. These findings suggest that oral P.g administration directly induced NAFLD in mice, which may be dependent on the ferroptosis of liver cells that occurs through the Th17/Treg imbalance induced by disordered microbial metabolism. Therefore, improving the periodontal environment is a novel treatment strategy for preventing NAFLD.


Refractory pediatric nonrhabdomyosarcoma soft tissue sarcoma associated with ectopic production of beta hCG and hypercalcemia induced by PTHrP.
A 3-month-old male with a mass on the right side of his back was admitted to our hospital. The tumor was a pathologically high-grade nonrhabdomyosarcoma soft tissue sarcoma (NRSTS). Treatment included subtotal tumor resection followed by chemotherapy. Elevation of serum beta hCG and hypercalcemia with detection of PTHrP was associated with tumor progression. The tumor was refractory to multiagent chemotherapy, and the patient died of the disease at 22 months of age. This case is novel in demonstrating a beta hCG secreting refractory NRSTS.


ATP-gated P2X7 receptor as a potential target for prostate cancer.
Prostate cancer is the most common malignancy of the male genitourinary system and is one of the leading causes of male cancer death. The P2X7 receptor is an important member of purine receptor family. It is a gated ion channel with adenosine triphosphate (ATP) as the ligand, which exists in a variety of immune tissues and cells and can be involved in tumorigenesis and tumor progression. Studies have shown that the P2X7 receptor is abnormally expressed in prostate cancer, and is related to the level of prostate-specific antigen, P2X7 receptor may be an early biomarker of prostate cancer. The P2X7 receptor is essential in the occurrence and development of prostate cancer. The P2X7 receptor mainly affects the invasion and metastasis of prostate cancer cells through epithelial mesenchymal transition/invasion-related genes and the PI3K/AKT and ERK1/2 signaling pathways. The P2X7 receptor could be a promising therapeutic target for prostate cancer.


Tuning of carbon dots emission color for sensing of Fe3+ ion and bioimaging applications.
Herein, we report a facile one-step synthetic strategy for fabrication of three (blue, green and yellow) fluorescent color carbon dots (CDs) from tomato (Solanum lycopersicum). The as-synthesized CDs showed emission peaks at 450, 520 and 560 nm for blue, green and yellow color CDs when excited at 370, 420 and 460 nm, respectively. Using tomato as a carbon source, the fabricated three fluorescent color CDs showed good water dispersity and high quantum yield. The analytical performances of three fluorescent color CDs are evaluated by detecting Fe3+ ion in biofluids and iron tablets. Upon the addition of Fe3+ ion under optimal conditions, the fluorescence intensity of three fluorescent color CDs was quenched linearly over the range of 0.1 to 2.0 μM. This method opens a new analytical strategy to quantify Fe3+ ion in iron tablets and biofluids with high sensitivity. Further, the uptake of three fluorescent color CDs into HeLa cells was confirmed by confocal laser scanning microscopy. Intracellular experiments demonstrated that the three fluorescent color CDs were effectively internalized the cells and show excellent biocompatibility and low toxicity, suggesting that the CDs can be used as good candidates for biomedical applications.


The effect of dextran and streptokinase on cerebral function and blood flow after cardiac arrest. An experimental study on the dog.
EEG activity and regional cerebral blood flow were studied during 5 h of maintenance following 12-16 min of cardiac arrest in 33 anesthetized dogs. Group I received no treatment. Group II received dextran 40. Group III were given both dextran 40 and streptokinase. In Group III the duration of flat EEG was significantly shorter than in Group I or II (P less than 0.002 and less than 0.008). The EEG score at 3 h had already reached a significant level as compared to Group I (P less than 0.005 at 3 h and P less than 0.002 at 5 h). The blood flow in the grey matter and hippocampus was greater in Group III than in Groups I and II. After 3 h the flow in Group III had already reached a significant level (P less than 0.0002, both grey matter and hippocampus) as compared to that in Group I. The findings suggest that impaired cerebral perfusion and function after cardiac arrest are most likely secondary to a combination of hemoconcentration, hyperviscosity, microthrombi formation, and edema at the microcirculatory level. Thus postischemic encephalopathy is in part due to perfusion abnormalities present after arrest, and is therefore amenable to therapy.


Molecular tools for the characterization of seizure susceptibility in genetic rodent models of epilepsy.
Epilepsy is a chronic neurological disorder characterized by abnormal neuronal activity that arises from imbalances between excitatory and inhibitory synapses, which are highly correlated to functional and structural changes in specific brain regions. The difference between the normal and the epileptic brain may harbor genetic alterations, gene expression changes, and/or protein alterations in the epileptogenic nucleus. It is becoming increasingly clear that such differences contribute to the development of distinct epilepsy phenotypes. The current major challenges in epilepsy research include understanding the disease progression and clarifying epilepsy classifications by searching for novel molecular biomarkers. Thus, the application of molecular techniques to carry out comprehensive studies at deoxyribonucleic acid, messenger ribonucleic acid, and protein levels is of utmost importance to elucidate molecular dysregulations in the epileptic brain. The present review focused on the great diversity of technical approaches available and new research methodology, which are already being used to study molecular alterations underlying epilepsy. We have grouped the different techniques according to each step in the flow of information from DNA to RNA to proteins, and illustrated with specific examples in animal models of epilepsy, some of which are our own. Separately and collectively, the genomic and proteomic techniques, each with its own strengths and limitations, provide valuable information on molecular mechanisms underlying seizure susceptibility and regulation of neuronal excitability. Determining the molecular differences between genetic rodent models of epilepsy and their wild-type counterparts might be a key in determining mechanisms of seizure susceptibility and epileptogenesis as well as the discovery and development of novel antiepileptic agents. This article is part of the Special Issue "NEWroscience 2018".


The role of focal-adhesion kinase in cancer - a new therapeutic opportunity.
Focal-adhesion kinase (FAK) is an important mediator of growth-factor signalling, cell proliferation, cell survival and cell migration. Given that the development of malignancy is often associated with perturbations in these processes, it is not surprising that FAK activity is altered in cancer cells. Mouse models have shown that FAK is involved in tumour formation and progression, and other studies showing that FAK expression is increased in human tumours make FAK a potentially important new therapeutic target.


Local Control Model of a Human Ventricular Myocyte: An Exploration of Frequency-Dependent Changes and Calcium Sparks.
Calcium (Ca2+) sparks are the elementary events of excitation-contraction coupling, yet they are not explicitly represented in human ventricular myocyte models. A stochastic ventricular cardiomyocyte human model that adapts to intracellular Ca2+ ([Ca2+]i) dynamics, spark regulation, and frequency-dependent changes in the form of locally controlled Ca2+ release was developed. The 20,000 CRUs in this model are composed of 9 individual LCCs and 49 RyRs that function as couplons. The simulated action potential duration at 1 Hz steady-state pacing is ~0.280 s similar to human ventricular cell recordings. Rate-dependence experiments reveal that APD shortening mechanisms are largely contributed by the L-type calcium channel inactivation, RyR open fraction, and [Ca2+]myo concentrations. The dynamic slow-rapid-slow pacing protocol shows that RyR open probability during high pacing frequency (2.5 Hz) switches to an adapted "nonconducting" form of Ca2+-dependent transition state. The predicted force was also observed to be increased in high pacing, but the SR Ca2+ fractional release was lower due to the smaller difference between diastolic and systolic [Ca2+]SR. Restitution analysis through the S1S2 protocol and increased LCC Ca2+-dependent activation rate show that the duration of LCC opening helps modulate its effects on the APD restitution at different diastolic intervals. Ultimately, a longer duration of calcium sparks was observed in relation to the SR Ca2+ loading at high pacing rates. Overall, this study demonstrates the spontaneous Ca2+ release events and ion channel responses throughout various stimuli.


The haemodynamic effects of intraaortic versus intravenous administration of protamine for reversal of heparin in man.
A recent report suggests that there may be advantages of intraaortic (IA) versus intravenous (IV) administration of protamine for reversal of heparin at the end of cardiopulmonary bypass. Because complete haemodynamic measurements were not performed in that study, we prospectively and randomly assessed the effects of protamine sulphate administered via either route. Patients were studied in the period immediately following cardiopulmonary bypass. Heparin, 300 units . Kg-1, plus 150 units . Kg-1 when required, was administered to keep the activated clotting time greater than 400 sec. Protamine 3 mg . Kg-1 was injected over 30 sec. intravenously in the IV group (n = 5) and into the ascending aorta in the IA group (n = 5). There was a significant decrease in arterial blood pressure (28.6 per cent) from 109.2 +/- 5.6 mmHg (S.E.) systolic to 78.0 +/- 9.0 mmHg observed in the IA group one minute post protamine, which returned to baseline by 2.5 min (analysis of variance p less than 0.05). No significant hypotension was observed in the IV group. There was no significant change in heart rate, left atrial pressure, central venous pressure, systemic vascular resistance or cardiac index with either IA or IV protamine. In contrast to our recent study in pigs there was no significant change in pulmonary artery pressure, pulmonary vascular resistance or cardiac output with protamine. Results indicate that there are no haemodynamic benefits of IA vs IV injection of protamine.


Splenic macrophages: antigen presenting cells for T1-2 antigens.
Thymus-independent antigens type 2 (T1-2) such as DNP-Ficoll or DNP-hydroxyethyl starch are selectively retained in mice and rats by a distinct population of macrophages in the marginal zone of the white pulp of the spleen and in the marginal sinus and septa and medulla of lymph nodes. There is suggestive evidence that the antibody response to the haptenic determinants, which is rapid in onset and persists for many weeks, is largely made by a sessile subpopulation of B cells in the spleen marginal zone characterized by the presence of surface IgM but lacking IgD. Splenectomy in adult mice, rats and humans greatly diminishes and delays the antibody response to DNP-Ficoll, but has no such effect on the response to thymus-dependent, or (in mice) to T1-1 immunogens. In humans, if DNP-Ficoll had been administered prior to splenectomy, a normal antibody response was made when the antigen was readministered after splenectomy, indicating that once the responding B cells had been stimulated they were no longer confined to the spleen. Experiments in mice showed that after elimination of B cells by irradiation, responsiveness to T1-2 antigens could be restored by spleen cells isolated as single-cell suspensions provided that the architecture of the spleen remained, but could not be restored if the mice had been splenectomized. This implies that both the spleen B cells and the marginal zone macrophages are required.(ABSTRACT TRUNCATED AT 250 WORDS)


Failure of a multi-subunit recombinant leishmanial vaccine (MML) to protect dogs from Leishmania infantum infection and to prevent disease progression in infected animals.
We report results of a Phase III trial of the multi-subunit recombinant Leishmania polyprotein MML for the protection of dogs against infection by Leishmania infantum. The antigen, also known as Leish-111f, is the first antileishmanial human vaccine entered Phase I clinical testing. The study was performed in a leishmaniasis endemic area of southern Italy. Three groups of 15 Leishmania-free beagle dogs each, received 3 monthly injections with vaccines A (MML+MPL-SE adjuvant), B (sterile saline = control) and C (MML+Adjuprime adjuvant), respectively, before transmission season 2002. The surviving dogs received a second three-dose vaccine course 1 year later. The dogs were naturally exposed to sandfly bites for 2.5 months in 2002, and for 5 months in 2003. Every 2 months post vaccination, dogs were examined by clinical and immunological evaluation, and by specific serology, microscopy, culture and PCR. A weak lymphoproliferative response to MML was seen in A and C groups throughout the study period. One year after the first vaccine course, the cumulative incidence of leishmanial infections was 40% in group A, 43% in group B and 36% in group C. Two-year post-vaccination (1 year after the second vaccine course) the cumulative incidence was 87% in group A (with three symptomatic cases), 100% in group B (with no symptomatic cases) and 100% in group C (with two symptomatic cases). The efficacy of the MML vaccine as an immunotherapeutic agent for the prevention of disease progression (subpatent infection-->asymptomatic patent infection-->symptomatic patent infection) was evaluated through follow-up of dogs found infected prior to the second vaccination. Among 15 infected animals, progression to a subsequent stage of infection was found in 5/6 dogs of group A, 3/6 of group B and 2/3 of group C. We conclude that vaccination with MML is not effective to prevent leishmaniasis infection and disease progression in dogs under field conditions.


Dysplastic enamel in odontomas.
A series of odontomas was studied by light microscopy, with special emphasis on abnormal enamel in these lesions. The findings were compared to those in amelogenesis imperfecta and in other conditions in which abnormal enamel is found. It was concluded that dysplastic enamel exhibits a number of appearances which are described in this article. A strongly basophilic laminated material was also found, often covering prismatic enamel, but it did not itself appear to be a form of enamel; its identity is not clear. Odontomas, being readily available, offer a good opportunity to study dysplastic enamel.


Precise analysis of the concentrations and isotopic compositions of molybdenum and tungsten in geochemical reference materials.
Molybdenum (Mo) is a redox-sensitive element and its concentrations and stable isotope compositions are widely used as a redox proxy in paleoceanography. Tungsten (W) is an emerging new isotope proxy, which has potential as a tracer for hydrothermal and early diagenetic processes. We present a new method for the precise and accurate analysis of Mo and W concentrations and isotope compositions from one single sample aliquot, thus saving mass of a sample and making the results directly comparable without concerns related to analytical or natural sample heterogeneity. After acid digestion, Mo and W are separated from the sample matrix using chelating resin NOBIAS Chelate-PA1 and anion exchange resin AG1 X8. Matrix removal is highly efficient: the remaining percentage is 10-2 to 10-5% with respect to the initial weight. Subsequently, samples are measured for Mo and W concentrations and isotope compositions using multi-collector inductivity coupled plasma mass spectrometry (MC-ICP-MS). For mass bias correction and determination of concentrations, we use standard-sample bracketing and in addition an external correction method employing ruthenium (Ru) for Mo and rhenium (Re) for W. This double correction approach results in an external reproducibility of or below 0.10‰ (2SD) for δ98Mo and 0.05‰ for δ186W based on ICP standard solutions (NIST SRM 3134 lot No. 130418 for Mo and NIST SRM 3163 lot No. 080331 for W). We present data for Mo and W in 12 geological reference materials including igneous rocks, sedimentary rocks, marine sediments, and manganese nodules. For Mo our method reproduces published values for the geological standard materials within analytical error of published values. For W, although published data do not always agree for a given geological standard material, our data agree within error with more recent data. We interpret a cause of the deviations is due to unknown effects of a desolvating nebulizer for MC-ICP-MS.


Preliminary survey of extended-spectrum β-lactamases (ESBLs) in nosocomial uropathogen Klebsiella pneumoniae in north-central Iran.
Infections caused by extended-spectrum β-lactamases (ESBLs) producing bacteria, including Klebsiella pneumoniae have increasingly subjected to therapeutic limitations and patients with these infections are at high risk for treatment failure, long hospital stays, high health care costs, and high mortality. The aim of this study was to screen the prevalence of the bla TEM,bla CTX-M and bla SHV ESBL genes in K. pneumoniae strains isolated from nosocomial urinary tract infections (UTIs). During the March 2016 to December 2017, one hundred isolates of K. pneumoniae were collected from urine specimens of patients suffering from nosocomial UTI referred to Khatam Al-Anbia hospital in Shahrud, north-central Iran. All isolates were identified by standard bacteriological tests. The pattern of antibiotic susceptibility was determined according to the CLSI guidelines. The presence of the ESBLs was investigated using the double-disc synergy test (DDST). Polymerase chain reaction technique was used to detect the bla TEM, bla CTX-M and bla SHV genes in DDST positive isolates. Most isolates showed remarkable resistance to tested antibiotics with highest rate against nitrofurantoin (75%) and trimethoprim/sulfamethoxazole (65%). The imipenem was the most effective antibiotic against K. pneumoniae isolates. ESBL phenotype was detected in 50 (50%) of isolates. The prevalence of bla TEM, bla CTX-M and bla SHV genes among 50 ESBLs-positive isolates was 25 (50%), 15 (30%) and 35 (70%) respectively. The bla TEM and bla SHV genes were seen in 25 isolates (50%) simultaneously. The findings of this study indicated the 50% frequency rate of ESBL-producing K. pneumoniae in our geographic region. Since the treatment of infections caused by this bacterium is associated with many limitations, this high prevalence is a warning sign to adopt new control policies to prevent further spread of this microorganism.


Near-present and future distribution of Anopheles albimanus in Mesoamerica and the Caribbean Basin modeled with climate and topographic data.
Anopheles albimanus is among the most important vectors of human malaria in Mesoamerica and the Caribbean Basin (M-C). Here, we use topographic data and 1950-2000 climate (near present), and future climate (2080) layers obtained from general circulation models (GCMs) to project the probability of the species' presence, p(s), using the species distribution model MaxEnt. The projected near-present distribution parameterized with 314 presence points related well to the known geographic distribution in the study region. Different model experiments suggest that the range of An. albimanus based on near-present climate surfaces covered at least 1.27 million km² in the M-C, although 2080 range was projected to decrease to 1.19 million km². Modeled p(s) was generally highest in Mesoamerica where many of the original specimens were collected. MaxEnt projected near-present maximum elevation at 1,937 m whereas 2080 maximum elevation was projected at 2,118 m. 2080 climate scenarios generally showed increased p(s) in Mesoamerica, although results varied for northern South America and no major range expansion into the mid-latitudes was projected by 2080. MaxEnt experiments with near present and future climate data suggest that An. albimanus is likely to invade high-altitude (>2,000 m) areas by 2080 and therefore place many more people at risk of malaria in the M-C region even though latitudinal range expansion may be limited.


Voltage dependence of the Ca2+-activated K+ conductance of human red cell membranes is strongly dependent on the extracellular K+ concentration.
The conductance of the Ca2+-activated K+ channel (gK(Ca)) of the human red cell membrane was studied as a function of membrane potential (Vm) and extracellular K+ concentration ([K+]ex). ATP-depleted cells, with fixed values of cellular K+ (145 mM) and pH (approximately 7.1), and preloaded with approximately 27 microM ionized Ca were transferred, with open K+ channels, to buffer-free salt solutions with given K+ concentrations. Outward-current conductances were calculated from initial net effluxes of K+, corresponding Vm, monitored by CCCP-mediated electrochemical equilibration of protons between a buffer-free extracellular and the heavily buffered cellular phases, and Nernst equilibrium potentials of K ions (EK) determined at the peak of hyperpolarization. Zero-current conductances were calculated from unidirectional effluxes of 42K at (Vm-EK) approximately equal to 0, using a single-file flux ratio exponent of 2.7. Within a [K+]ex range of 5.5 to 60 mM and at (Vm-EK) greater than or equal to 20 mV a basic conductance, which was independent of [K+]ex, was found. It had a small voltage dependence, varying linearly from 45 to 70 microS/cm2 between 0 and -100 mV. As (Vm-EK) decreased from 20 towards zero mV gK(Ca) increased hyperbolically from the basic value towards a zero-current value of 165 microS/cm2. The zero-current conductance was not significantly dependent on [K+]ex (30 to 156 mM) corresponding to Vm (-50 mV to 0). A further increase in gK(Ca) symmetrically around EK is suggested as (Vm-EK) becomes positive. Increasing the extracellular K+ concentration from zero and up to approximately 3 mM resulted in an increase in gK(Ca) from approximately 50 to approximately 70 microS/cm2. Since the driving force (Vm-EK) was larger than 20 mV within this range of [K+]ex this was probably a specific K+ activation of gK(Ca). The Ca2+-activated K+ channel of the human red cell membrane is an inward rectifier showing the characteristic voltage dependence of this type of channel.


Adult neurogenesis in eight Megachiropteran species.
The present study evaluated, using immunohistochemical methods, the presence and characteristics of proliferating and newly generated neurons in the brain of eight wild-caught adult Megachiropteran species. For the neurogenic patterns observed, direct homologies are evident in other mammalian species; however, there were several distinctions in the presence or absence of proliferating and immature neurons, and migratory streams that provide important clues regarding the use of the brain in the analysis of Chiropteran phylogenetic affinities. In all eight species studied, numerous Ki-67- and doublecortin (DCX)-immunopositive cells were identified in the subventricular zone (SVZ). These cells migrated to the olfactory bulb through a Primate-like rostral migratory stream (RMS) that is composed of dorsal and ventral substreams which merge before entering the olfactory bulb. Some cells were observed emerging from the RMS coursing caudally and dorsally to the rostral neocortex. In the dentate gyrus of all species, Ki-67- and DCX-expressing cells were observed in the granular cell layer and hilus. Similar to Primates, proliferating cells and immature neurons were identified in the SVZ of the temporal horn of Megachiropterans. These cells migrated to the rostral and caudal piriform cortex through a Primate-like temporal migratory stream. Sparsely distributed Ki-67-immunopositive, but DCX-immunonegative, cells were identified in the tectum, brainstem and cerebellum. The observations from this study add to a number of neural characteristics that phylogenetically align Megachiropterans to Primates.


Epithelioid hemangioendothelioma: cytomorphology and histological features of a case.
A case of epithelioid hemangioendothelioma involving the left lung, the 10th thoracic vertebra, and paravertebral soft tissues is presented. Preoperative sputum cytology was thought to show adenocarcinoma. Histologic examination demonstrated an intravascular bronchioloalveolar tumor; immunohistochemical staining for factor VIII-related antigen revealed strongly positive tumor cells. The cytomorphology of this rare neoplasm is discussed.


Measurement Error, Reliability, and Minimum Detectable Change in the Mini-Mental State Examination, Montreal Cognitive Assessment, and Color Trails Test among Community Living Middle-Aged and Older Adults.
Knowing the reliability of cognitive tests, particularly those commonly used in clinical practice, is important in order to interpret the clinical significance of a change in performance or a low score on a single test. To report the intra-class correlation (ICC), standard error of measurement (SEM) and minimum detectable change (MDC) for the Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), and Color Trails Test (CTT) among community dwelling older adults. 130 participants aged 55 and older without severe cognitive impairment underwent two cognitive assessments between two and four months apart. Half the group changed rater between assessments and half changed time of day. Mean (standard deviation) MMSE was 28.1 (2.1) at baseline and 28.4 (2.1) at repeat. Mean (SD) MoCA increased from 24.8 (3.6) to 25.2 (3.6). There was a rater effect on CTT, but not on the MMSE or MoCA. The SEM of the MMSE was 1.0, leading to an MDC (based on a 95% confidence interval) of 3 points. The SEM of the MoCA was 1.5, implying an MDC95 of 4 points. MoCA (ICC = 0.81) was more reliable than MMSE (ICC = 0.75), but all tests examined showed substantial within-patient variation. An individual's score would have to change by greater than or equal to 3 points on the MMSE and 4 points on the MoCA for the rater to be confident that the change was not due to measurement error. This has important implications for epidemiologists and clinicians in dementia screening and diagnosis.


Predictability of the effects of epidural injection in lumbar spinal stenosis by assessment of lumbar MRI scans.
Numerous classification systems have been proposed to interpret lumbar MRI scans. The clinical impact of the measured parameters remains unclear. To evaluate the clinical significance of imaging results in patients with multisegmental degenerative pathologies, treating specialists can perform image-guided local injections to target defined areas such as the epidural space. The aim of this retrospective study was to evaluate the correlation between lumbar spinal stenosis measurements obtained by MRI and improvement obtained through local epidural injection. In this retrospective study various measurement and classification systems for lumbar spinal stenosis were applied to MRI scans of 100 patients with this pathological condition. The reported effect of epidural bupivacaine/triamcinolone injections at the site was recorded in these patients and a comparative analysis performed. MRI features assessed in this study did not show any relevant correlation with reported pain relief after epidural injection in patients with chronic lumbar stenosis, with the exception of posterior disc height with a weak Kendall's tau of -0.187 (p= 0.009). Although MRI is crucial for evaluating lumbar spinal stenosis, it cannot replace but is rather complementary to a good patient history and clinical examination or the results of local diagnostic injections.


A missed medial humeral epicondyle fracture with incarcerated fragment in the elbow joint and ulnar nerve palsy: A rare case report.
Medial epicondyle fracture associated with incarcerated intra-articular fragment and ulnar nerve palsy is uncommon and frequently missed. We report a case of 13-year-old boy with incarcerated medial epicondyle fracture fragment in ulnohumeral joint and ulnar nerve palsy, which was managed successfully by open reduction internal fixation and ulnar nerve transposition.


The properties of hydrogenase from Thiocapsa roseopersicina.
The soluble and chromatophore-bound hydrogenases from the purple sulphur bacterium Thiocapsa roseopersicina strain BBS were purified up to homogeneity and the properties studied. The amino acid composition of hydrogenase preparations from different fractions of T. roseopersicina is identical, glycine and alanine as N-terminal amino acid residues. In comparison with other hydrogenases, especially in the immobilized state, the preparations obtained are shown to be more stable to O2 during storage and they are characterized by high thermal stability. Inactivation is observed above 78--80 degrees C and the optimal temperature for enzyme action is 70 degrees C. The homogeneous enzyme preparations catalyse the exchange reaction between 2H2 and H2O and reversible redox reactions of methyl viologen and benzyl viologen as well as H2 formation from reduced ferredoxin. According to our data, the hydrogenase of T. roseopersicina bound with chromatophores is identical to the soluble one.


Substantial Cardiovascular Morbidity in Adults With Lower-Complexity Congenital Heart Disease.
Although lower-complexity cardiac malformations constitute the majority of adult congenital heart disease (ACHD), the long-term risks of adverse cardiovascular events and relationship with conventional risk factors in this population are poorly understood. We aimed to quantify the risk of adverse cardiovascular events associated with lower-complexity ACHD that is unmeasured by conventional risk factors. A multitiered classification algorithm was used to select individuals with lower-complexity ACHD and individuals without ACHD for comparison among >500 000 British adults in the UK Biobank. ACHD diagnoses were subclassified as isolated aortic valve and noncomplex defects. Time-to-event analyses were conducted for the primary end points of fatal or nonfatal acute coronary syndrome, ischemic stroke, heart failure, and atrial fibrillation and a secondary combined end point for major adverse cardiovascular events. Maximum follow-up time for the study period was 22 years with retrospectively and prospectively collected data from the UK Biobank. We identified 2006 individuals with lower-complexity ACHD and 497 983 unexposed individuals in the UK Biobank (median age at enrollment, 58 [interquartile range, 51-63] years). Of the ACHD-exposed group, 59% were male, 51% were current or former smokers, 30% were obese, and 69%, 41%, and 7% were diagnosed or treated for hypertension, hyperlipidemia, and diabetes mellitus, respectively. After adjustment for 12 measured cardiovascular risk factors, ACHD remained strongly associated with the primary end points, with hazard ratios ranging from 2.0 (95% CI, 1.5-2.8; P<0.001) for acute coronary syndrome to 13.0 (95% CI, 9.4-18.1; P<0.001) for heart failure. ACHD-exposed individuals with ≤2 cardiovascular risk factors had a 29% age-adjusted incidence rate of major adverse cardiovascular events, in contrast to 13% in individuals without ACHD with ≥5 risk factors. Individuals with lower-complexity ACHD had a higher burden of adverse cardiovascular events relative to the general population that was unaccounted for by conventional cardiovascular risk factors. These findings highlight the need for closer surveillance of patients with mild to moderate ACHD and further investigation into management and mechanisms of cardiovascular risk unique to this growing population of high-risk adults.


Case report: A rare case of eccrine carcinoma.
Sweat gland carcinoma is very rare with a reported incidence of less than 0.005% of all tumour specimens resected surgically (Tulenko and Conway, 1965) [1]. A sixty year old male patient presented to us with a solitary swelling over the left chest wall since two months. Cutaneous apocrine gland carcinoma, a subtype of sweat gland carcinoma, is a very rare malignant neoplasm arising in areas of high apocrine sweat gland density. The variability of the histological features even in the same tumour, and its rarity, have contributed to some confusion regarding the classification of sweat gland carcinoma. Sweat gland carcinomas are a rare group of tumours with potential for local destruction as well as distant metastasis. Wide surgical excision along with regional lymph node dissection in the presence of clinically positive nodes is the recommended treatment. However, a frequent follow up is essential to detect early recurrence as well as distant metastasis.


Androgen receptor inclusions acquire GRP78/BiP to ameliorate androgen-induced protein misfolding stress in embryonic stem cells.
Commitment of differentiating embryonic stem cells (ESCs) toward the various lineages is influenced by many factors, including androgens. However, the mechanisms underlying proteotoxic stress conferred by androgen receptor (AR) actions on embryonic cell fate remains unclear. Here we show that mouse ESCs display stress-related cellular phenotypes in response to androgens during early phase of differentiation. Androgen induced a significant increase in the percentage of ESCs and embryoid bodies with the intranuclear and juxtanuclear AR inclusions, which were colocalized with the E3 ubiquitin ligase, C terminus of Hsc70-interacting protein. Caspase-3 activity corresponded with AR expression, was enhanced in cells engaged more differentiation phenotypes. Androgen-mediated accumulation of AR aggregates exacerbated endoplasmic reticulum (ER) stress and rendered ESCs susceptible to apoptosis. Increasing expression levels of the ER chaperones, GRP78/BiP and GRP94, as well as ER stress markers, such as ATF6, phosphorylated PERK, GADD153/CHOP and spliced XBP-1 mRNA, were dramatically elevated in ESCs overexpressing AR. We found that androgen induced GRP78/BiP to dissociate from ATF6, and act as an AR-interacting protein, which was recruited into AR inclusions in ESCs. GRP78/BiP was also colocalized with AR inclusions in the cells of spinal bulbar muscular atrophy transgenic mouse model. Overexpression of GRP78/BiP suppressed ubiquitination of AR aggregates and ameliorated the misfolded AR-mediated cytopathology in ESCs, whereas knockdown of GRP78/BiP increased the accumulation of AR aggregates and significantly higher levels of caspase-3 activity and cell apoptosis. These results generate novel insight into how ESCs respond to stress induced by misfolded AR proteins and identify GRP78/BiP as a novel regulator of the AR protein quality control.


Genetic control of mitogen-induced B-cell hyperproliferation in SM/J mice.
Previous studies have shown that B cells from SM/J mice exhibit hyperproliferative responsiveness to several bacterial-derived B-cell mitogens. This hyperresponse trait was found to be under autosomal, polygenic control by non-H-2 genes. We have now estimated the number of genes involved by statistical analysis of the proliferative responses of splenocytes from SM/J and low-responder C57BL/6J strains, and progeny from the (B6 X SM)F1, F2 and (F1 X B6) crosses. The number of loci involved was ascertained using two different statistical approaches. An estimate of two loci was determined using chi-squared statistics. The second approach, based on an additive model in the natural log scale, also pointed to a lower bound of two genes. We conclude that the hyperresponse to B-cell mitogens in SM/J mice is determined by two autosomal genes which are not linked to the H-2 major histocompatibility complex.


LncRNA TDRG1 promotes the metastasis of NSCLC cell through regulating miR-873-5p/ZEB1 axis.
Long noncoding RNAs (lncRNAs) are dysregulated in various malignancies and involved in the growth and aggressive phenotypes of cancer cells. Previous studies indicate that lncRNA testis development related 1 (TDRG1) plays critical roles in the development of several malignancies. Nevertheless, the molecular mechanism underlying TDRG1 contributes to non-small cell lung cancer (NSCLC) remains elusive. Here, we demonstrate that TDRG1 is significantly overregulated in NSCLC tissues and cell lines. Knockdown of TDRG1 inhibits the proliferation and metastatic-related traits of NSCLC cell in vitro whereas overexpression of TDRG1 causes opposite results. In addition, TDRG1 silencing inhibits the growth and metastatic ability of NSCLC cell in vivo as demonstrated by xenograft tumor model and lung metastasis model. The binding capacity of TDRG1 with miR-873-5p is demonstrated by bioinformatics prediction tool and luciferase reporter gene assay. Additional, the rescue experiments indicate that TDRG1 interacts with miR-873-5p and its expression is positively associated with the target of miR-873-5p, zinc finger e-box binding homeobox 1 (ZEB1). Altogether, lncRNA TDRG1 facilitates the progression of NSCLC via interacting with miR-873-5p and positively regulates the expression of ZEB1.


Neurophysiological changes during shortening osteotomies of the spine.
Kyphotic deformities with sagittal imbalance of the spine can be treated with spinal osteotomies. Those procedures are known to have a high incidence of neurological complications, in particular at the thoracic level. Motor evoked potentials (MEPs) have been widely used in helping to avoid major neurological deficits postoperatively. Previous reports have shown that a significant proportion of such cases present with important transcranial MEP (Tc-MEP) changes during surgery with some of them being predictive of postoperative deficits. Our aim was to study Tc-MEP changes in a consecutive series of patients and correlate them with clinical parameters and radiological changes. Retrospective case notes study from a prospective patient register. Eighteen patients undergoing posterior shortening osteotomies (nine at thoracic and nine at lumbar levels) for kyphosis of congenital, degenerative, inflammatory, or post-traumatic origin were included. Loss of at least 80% of Tc-MEP signal expressed as the area under the curve percentual change, of at least one muscle. We studied the relation between outcome measure (80% Tc-MEP loss in at least one muscle group) and amount of posterior vertebral body shortening as well as angular correction measured on computed tomography scans, occurrence of postoperative deficits, intraoperative blood pressure at the time of the osteotomy, and hemoglobin (Hb) change. All patients showed significant Tc-MEP changes. In particular, greater than 80% MEP loss in at least one muscle group was observed in five of nine patients in the thoracic group and four of nine patients in the lumbar group. No surgical maneuver was undertaken as a result of this loss in an effort to improve motor responses other than verifying the stability of the construct and the extent of the decompression. Four patients developed postoperative deficits of radicular origin, three of them recovering fully at 3 months. No relation was found between intraoperative blood pressure, Hb changes, and Tc-MEP changes. Severity of Tc-MEP loss did not correlate with postoperative deficits. Shortening of more than 10 mm was linked to more severe Tc-MEP changes in the thoracic group. Transcranial MEP changes during spinal shortening procedures are common and do not appear to predict severe postoperative deficits. Total loss of Tc-MEP (not witnessed in our series) might require a more drastic approach with possible reversal of the correction and wake-up test.


Involvement of tumor necrosis factor alpha in intestinal epithelial cell proliferation following Paneth cell destruction.
An intravenous injection of diphenylthiocarbazone (dithizone), a zinc chelator, induces selective killing of Paneth cells which have a large amount of zinc in their cytoplasmic granules. A transient wave of intestinal epithelial cell proliferation occurs at 12 h after the injection. Paneth cells have tumor necrosis factor (TNF)-alpha protein in their cytoplasmic granules, and TNF-alpha has a proliferative effect on intestinal epithelial cells in vitro. The aim of this study is to clarify the in vivo role of TNF-alpha in intestinal epithelial cell proliferation using a dithizone-treated rat model. Male Wistar rats received a dithizone (100 mg/kg of body weight) injection with or without TNF-alpha inhibitor, pentoxifylline (100 mg/kg), neutralizing anti-TNF-alpha antibody (2 mg/kg), or nuclear transcription factor kappaB (NF-kappaB) inhibitors: pyrrolidine dithiocarbamate (100 mg/kg) or N-acetyl-L-cystein (100 mg/kg). The activation of NF-kappaB was examined by the electrophoretic mobility shift assay, and cellular proliferation by BrdU labeling. Without any inhibitors, dithizone treatment evoked NF-kappaB activation in the ileal mucosa with its peak level at 2 h after the injection. TNF-alpha inhibition reduced the NF-kappaB activation, and blocked a transient wave of epithelial cell proliferation 12 h after the injection. NF-kappaB inhibitors also reduced the NF-kappaB activation and epithelial cell proliferation. TNF-alpha released from degenerated Paneth cells was, in part, responsible for the intestinal cell proliferation through the activation of NF-kappaB, suggesting its proliferative effect on intestinal epithelial cells.


The human cathepsin D-encoding gene is transcribed from an estrogen-regulated and a constitutive start point.
The 5' flanking sequences, exon 1 and part of intron 1 of the human cathepsin D (CTD)-encoding gene (CTD) have been cloned and sequenced. RNase protection experiments identified two major transcription start points (tsp) located 14 and 63 nucleotides upstream of the start codon. The proximal -14, but not the distal -63 tsp has upstream near-concensus TATAAA and CCAAT sequences. Estrogens increase transcription from the -14 tsp, but not the -63 tsp and CTD is therefore unique among estrogen-regulated genes in having estrogen-regulated and constitutive transcription. Sequencing approximately 800 bp upstream and 600 bp downstream of the tsp failed to identify a consensus 13-bp palindromic estrogen-response element (ERE); however, four half-palindrome GGTCA motifs were located within 340 bp upstream of the -14 bp tsp. Thus, estrogen regulation of CTD may not be mediated by a consensus ERE.


Effects of particle size, organic matter and ionic strength on the phosphate sorption in different trophic lake sediments.
The trophic status and development of lake system is significantly influenced by the phosphate sorption at the sediment-water interface. The effects of organic matter, particle size and ionic strength on the phosphate sorption by sediments were investigated in this study. The results show that maximum phosphate sorption capacity (Qmax), equilibrium phosphate concentration (EPC0), phosphate sorption efficiency (k) and phosphate sorption rate decreased as particle size increased for all the studied sediments. But the reliable desorbed phosphorus (RDP) increased. Qmax, EPC0, k and RDP of different particle size fractions varied and there were no obvious differences among different trophic lake sediments. Qmax, EPC0 increased while k and RDP decreased as the pollution level of lake sediment increased. The phosphate sorption mainly occurred within 0.5 h. Power function and simple Elovich models were the best kinetic models for the phosphate sorption of the different particle size fractions. Qmax and phosphate sorption rate decreased with the decreasing of organic matter content of sediments and the increasing of ionic strength. This study suggests that ionic strength and organic matter had similar effects on the phosphate sorption for different trophic lake sediments.


Changes in plasma and colonic mucosa fatty acid profiles in rats with ulcerative colitis induced by trinitrobenzene sulfonic acid.
Polyunsaturated fatty acids have a key role in the pathogenesis of inflammatory bowel disease since some of the arachidonic acid-derived eicosanoids have been found to be increased in inflamed intestinal mucosa in the acute phase of human disease. The aim of this study was to prospectively assess plasma and colon mucosa fatty acid patterns in rats with experimental ulcerative colitis. Twenty rats were treated with trinitrobenzene sulfonic acid and 20 with NaCl; two groups were killed after one week and two after two weeks to evaluate colon damage. Plasma was obtained by aortic puncture and colonic mucosa was scraped off and the fatty acid pattern was determined by gas-liquid chromatography. Total, saturated, and monounsaturated plasma fatty acids were significantly higher in both periods of ulcerative colitis as compared to controls. Plasma n-6 fatty acids were increased after treatment, but no significant changes were observed concerning to n-3 fatty acids. With regard to colon mucosa, saturated and monounsaturated fatty acids did not change because of the disease; however, n-6 fatty acids decreased in the first week and increased in the second week and n-3 fatty acids were increased. Changes on the fatty acid distribution in plasma did not parallel to those of colonic mucosa except for 22:6(n-3). We have also found that experimental ulcerative colitis induced by trinitrobenzene sulfonic acid reproduces many of the features related to changes in plasma and colon mucosa fatty acids observed in the human disease.


Spatial and temporal genetic variability in French populations of the peach-potato aphid, Myzus persicae.
The peach-potato aphid, Myzus persicae (Sulzer), has a complex reproductive mode in which some lineages reproduce by continuous parthenogenesis, whereas others reproduce sexually once a year. The climate is thought to act directly on the reproductive mode, because sexual eggs are the only form that can resist frost in cold regions. Sexual reproduction necessitates an obligatory host alternation that may result in long-distance dispersal. Here, we examined the genetic variability at seven microsatellite loci of populations of M. persicae in France, where both reproductive modes occur. We provide clear genetic evidence that the breeding system affects genotypic variability, as cyclically parthenogenetic aphids are far more variable than their obligately parthenogenetic counterparts. A temporal decrease in genetic variability and a temporal genetic differentiation effect suggest the existence of selective factors that play an important role in shaping the genetic structure of M. persicae populations. Lastly, differences in the population structure between reproductive modes suggest that the migration associated with the change of host during sexual reproduction lowers the level of population differentiation.


Author Correction: Efficient electrical detection of mid-infrared graphene plasmons at room temperature.
In the version of this Article originally published, the units of the right-hand y axis of Fig. 2a were incorrectly labelled as mS; they should have been µS. Also, the x-axis tick marks of Fig. 3b should have been aligned with Fig. 3a,c. These have now been corrected.


[Effect of vitamin D2 on the monooxygenase activity of the rat liver].
Stimulation of lipid peroxidation in the liver microsomal fraction and alteration in the xenobiotic metabolism were found within twelve hrs after a single vitamin D2 administration (960,000 MU/kg) into male rats. The effects observed appear to occur due to the vitamin D2 prooxidative properties.


Influence of oral comprehensive nursing intervention on mechanically ventilated patients in ICU: a randimized controlled study.
To explore the effect of oral comprehensive nursing intervention on mechanically ventilated patients in ICU. Select 76 cases of mechanically ventilated patients in severe ICU admitted to our hospital from January 2022 to October 2022 as the research objects, and divide them into the control group and the observation group according to the way the patients receive oral care. 38 cases each. The patients in the control group received routine nursing intervention, and the patients in the observation group received comprehensive oral nursing intervention on the basis of the nursing of the control group. The clinical index data, oropharyngeal hygiene, pH value, blood gas analysis index levels, and the occurrence and death of ventilator-associated pneumonia were compared between the two groups of patients. The hospitalization time of the two groups was compared (P > 0.05); the mechanical ventilation time and ICU stay time of the observation group were significantly lower than those of the control group (all, P < 0.05); the oral odor scores, The plaque index and soft scale index were significantly lower than those of the control group (all, P < 0.05); the pH value, PaO 2 value, and SpO 2 value of the observation group were significantly lower than those of the control group, and the PaCO 2 value was significantly higher than that of the control group. group (all, P < 0.05); the incidence of VAP in the control group was 55.26%, and the mortality rate was 15.79%, the incidence rate of VAP in the observation group was 21.05%, and the mortality rate was 2.63%, and the incidence rate and mortality rate of VAP in the observation group were significantly lower in the control group (all, P < 0.05). The application of nursing intervention can effectively promote the recovery of patients, improve the hygiene of patients' oropharynx, adjust the levels of pH and blood gas-related indicators in patients, and reduce VAP in patients. risk of morbidity and mortality.


Evaluation of Individualized Pain Plans for Children With Sickle Cell Disease Admitted for Vaso-occlusive Crisis at Riley Hospital for Children.
Vaso-occlusive crisis (VOC) is the most common problem reported by patients with sickle cell disease (SCD). The objective of this study was to evaluate the impact of individualized pain plans in pediatric patients with SCD admitted for VOC. This was a pre- and post-study of patients with SCD admitted to Riley Hospital for Children for VOC from July 1, 2019, through July 1, 2020. The primary outcome was length of inpatient stay for VOC. Secondary outcomes included final pain score, days on scheduled opioids, days on breakthrough opioids, and average morphine milligram equivalents (MME) used per day. Nine patients were included. The mean age was 16 years (range, 10-20 years). Key clinical findings were decreases in median [IQR] for final pain scores (7 [4.5-9] vs 6 [2.5-8], p = 0.396) and number of days of breakthrough opioid use (5 [3-8] vs 4 [2.5-5.5], p = 0.233). Following implementation of an individualized pain plan, there was an increase in median average MME per day (65.94 [53.1-97.7] vs 82.85 [41-114.3], p = 0.844). Median length of stay and days on scheduled opioids remained the same. This study demonstrated that use of individualized pain plans in a small population of patients with SCD might result in decreased pain scores and decreased days on breakthrough opioids.


Purification and properties of an arginyl aminopeptidase from Debaryomyces hansenii.
A metallo arginyl aminopeptidase (EC 126.96.36.199) activated by Co(2+) was isolated from Debaryomyces hansenii CECT 12487. The enzyme was purified after precipitation with protamine sulphate, followed by a weak anion exchange chromatography, gel filtration chromatography and a strong anion exchange chromatography. The arginyl aminopeptidase (AAP) was purified 337 folds, with a 18% recovery. The AAP appeared to be a dimer with a molecular mass of 101 kDa. The enzyme was active in the pH range from 6 to 9. The optimal activity was detected at pH 7.0 and at 37 degrees C. AAP activity was inhibited by typical aminopeptidase inhibitors (puromycin and bestatin), reducing agents (DTT), chelating agents (EDTA, EGTA and phenantroline) and sulphydryl groups reagents (iodoacetate). Ca(2+), Mn(2+) and Co(2+) activated the enzyme, while Cu(2+), Cd(2+), Hg(2+) and Mg(2+) inhibited it. The K(m) values calculated for Arg-AMC (7-amido-4-methylcoumarin) and Leu-AMC were 0.071 and 0.094 mM, respectively. The enzyme showed maximum specificity for basic amino acids (Arg and Lys), but was also able to hydrolyze non-charged amino acids (Leu, Met and Ala) and, at a minor rate, aromatic amino acids (Phe and Tyr). AAP showed higher activity when an acid residue was located at the C-terminal position of dipeptides. The described purification of an arginyl aminopeptidase from the yeast D. hansenii can contribute to the lack of knowledge about the exopeptidase activity in one of the yeasts more frequently isolated in sausage and to understand its role during the ripening of a fermented sausage.


Effect of polyamines on cell-free protein synthesizing systems from rat cerebral cortex, cerebellum and liver.
The naturally occurring polyamines spermine and spermidine stimulate in low concentrations the amino acid-incorporating activity of cell-free systems from cerebral cortex, cerebellum and liver from the rat to a significant degree, but in high concentrations they have strong inhibiting effects on these systems. Maximal polyamine stimulation is observed with suboptimal Mg2+ concentrations in the reaction mixtures; increasing Mg2+ to 15 mM results in an inhibition of cell-free protein synthesis by polyamines even at low concentrations. The stimulating effect of spermine and spermidine involves a facilitated binding of template RNA and aminoacylated tRNA to the ribosomes. In contrast, the aminoacylation of tRNA is not influenced. These data indicate that spermine as well as spermidine have regulatory functions during the translational process in cerebral cortex, cerebellum and liver. They seem to interact with ribosomal functions and to cause a shift in the optimal Mg2+ concentration required for the synthesis of brain and liver proteins.


Sulfone therapy in the treatment of leukocytoclastic vasculitis. Report of three cases.
Dapsone therapy for leukocytoclastic (necrotizing) vasculitis has been little used, except for the variant forms of erythema elevatum diutinum and urticarial vasculitis. We report three patients with the common (palpable purpura) form of the disease, limited to the skin, and successfully treated with moderate doses of dapsone (100-150 mg daily). Although the natural course of leukocytoclastic vasculitis is highly unpredictable, the prompt disappearance of new lesion formation after initiation of treatment and the rapid recurrence of lesions after therapy is discontinued (both often within 4 to 8 days after the critical dose level is reached) reflect drug efficacy. We believe that dapsone deserves wider evaluation as a therapeutic agent for chronic or recurrent cases of the common form of leukocytoclastic vasculitis.


Nonhyperoxic retrolental fibroplasia.
Retrolental fibroplasia (RLF) has most often been observed in preterm infants exposed to high ambient oxygen concentrations. This case report describes an infant near term who was never exposed to supplemental oxygen but in whom stage 4 RLF developed. The etiologic role of exchange transfusion is discussed.


REM sleep behavior disorder with predominant nightmares in a patient with ischemic pontine lesions.
Rapid eye movement (REM) sleep behavior disorder (RBD) is a parasomnia typically associated with synucleinopathy and evolving to neurodegenerative disorders. RBD is caused by impairment of brainstem nuclei controlling REM sleep muscle atonia. Rarely, focal lesions of the brainstem can cause secondary RBD. We present the case of a 74-year-old patient, previously evaluated at age 70 years for insomnia and periodic limb movements during sleep, who then rapidly developed unpleasant dreams with minor motor behavior, affecting his sleep quality. Polysomnography recorded REM sleep without atonia and motor behaviors in REM sleep. Ischemic lesions in the pons were detected by magnetic resonance imaging. Clinical, biological, and instrumental biomarkers of neurodegeneration were repeatedly negative at 2 years' follow-up. Although rare, a lesional cause of RBD must be considered in cases of atypical presentation and without evidence of neurodegeneration. The complaint of unpleasant dreams suggests a possible role of brainstem nuclei controlling REM sleep atonia in affecting oneiric content. Biscarini F, Montini A, Antelmi E, Vandi S, Pizza F, Plazzi G. REM sleep behavior disorder with predominant nightmares in a patient with ischemic pontine lesions. J Clin Sleep Med. 2022;18(3):945-948.


The effect of polyunsaturated phosphatidyl choline in the treatment of acute viral hepatitis.
Polyunsaturated phosphatidyl choline is a preparation often advocated for diseases of the liver. In a randomized open controlled trial, a preparation of polyunsaturated phosphatidyl choline, at a dose of 900 mg orally daily, was given to 22 patients with acute viral hepatitis. A control group of 25 patients was not treated. Serial serum bilirubin and alanine amino transferase levels were measured up to 12 weeks. The falls in their levels after 2 and 5 weeks, and the lengths of time to their normalization, were not significantly different in the treated group compared to the control group. The results indicated that polyunsaturated phosphatidyl choline had no beneficial effect on the course of acute viral hepatitis.


17beta-estradiol affects proliferation and apoptosis of rat prostatic smooth muscle cells by modulating cell cycle transition and related proteins.
Abundant evidence indicates that estrogens have an important role in the pathology of benign prostatic hyperplasia (BPH). To investigate the effect of 17beta-estradiol (E2) on the proliferation and apoptosis of prostatic smooth muscle cells (PSMCs), rat PSMCs were obtained and exposed to gradient concentrations (0.1-100 nmol/l) of E2 over varying amounts of time. The progression of cell cycle, cellular apoptosis, cyclin D1, Bcl-2 and Bax proteins were detected. The data show that the effect of E2 on rat PSMCs is bilateral: it promotes cell proliferation by enhancing the expression of cyclin D1, which accelerates G1 to S phase transition; on the other hand, it induces apoptosis of the cells by up-regulating the expression of Bax. We thus suggest that an increase in estrogen may exert a launching effect in the pathology of BPH.


A commercial line probe assay for the rapid detection of rifampicin resistance in Mycobacterium tuberculosis: a systematic review and meta-analysis.
Mycobacterium tuberculosis is a leading cause of death worldwide. In multi-drug resistant tuberculosis (MDR-TB) infectiousness is frequently prolonged, jeopardizing efforts to control TB. The conventional tuberculosis drug susceptibility tests are sensitive and specific, but they are not rapid. The INNO-LiPA Rif. TB (LiPA) is a commercial line probe assay designed to rapidly detect rifampicin resistance, a marker of MDR-TB. Although LiPA has shown promising results, its overall accuracy has not been systematically evaluated. We did a systematic review and meta-analysis to evaluate the accuracy of LiPA for the detection of rifampicin-resistant tuberculosis among culture isolates and clinical specimens. We searched Medline, Embase, Web of Science, BIOSIS, and Google Scholar, and contacted authors, experts and the manufacturer. Fifteen studies met our inclusion criteria. Of these, 11 studies used culture isolates, one used clinical specimens, and three used both. We used a summary receiver operating characteristic (SROC) curve and Q* index to perform meta-analysis and summarize diagnostic accuracy. Twelve of 14 studies that applied LiPA to isolates had sensitivity greater than 95%, and 12 of 14 had specificity of 100%. The four studies that applied LiPA directly to clinical specimens had 100% specificity, and sensitivity that ranged between 80% and 100%. The SROC curve had an area of 0.99 and Q* of 0.97. LiPA is a highly sensitive and specific test for the detection of rifampicin resistance in culture isolates. The test appears to have relatively lower sensitivity when used directly on clinical specimens. More evidence is needed before LiPA can be used to detect MDR-TB among populations at risk in clinical practice.


Comparative effectiveness of aspirin for symptomatic venous thromboembolism prophylaxis in patients undergoing total joint arthroplasty, a cohort study.
This study compares the symptomatic 90-day venous thromboembolism (VTE) rates in patients receiving aspirin to patients receiving low-molecular weight heparin (LMWH) or direct oral anticoagulants (DOACs), after total hip (THA) and total knee arthroplasty (TKA). Data were collected from a multi-centre cohort study, including demographics, confounders and prophylaxis type (aspirin alone, LMWH alone, aspirin and LMWH, and DOACs). The primary outcome was symptomatic 90-day VTE. Secondary outcomes were major bleeding, joint related reoperation and mortality within 90 days. Data were analysed using logistic regression, the Student's t and Fisher's exact tests (unadjusted) and multivariable regression (adjusted). There were 1867 eligible patients; 365 (20%) received aspirin alone, 762 (41%) LMWH alone, 482 (26%) LMWH and aspirin and 170 (9%) DOAC. The 90-day VTE rate was 2.7%; lowest in the aspirin group (1.6%), compared to 3.6% for LMWH, 2.3% for LMWH and aspirin and 2.4% for DOACs. After adjusted analysis, predictors of VTE were prophylaxis duration < 14 days (OR = 6.7, 95% CI 3.5-13.1, p < 0.001) and history of previous VTE (OR = 2.4, 95% CI 1.1-5.8, p = 0.05). There were no significant differences in the primary or secondary outcomes between prophylaxis groups. Aspirin may be suitable for VTE prophylaxis following THA and TKA. The comparatively low unadjusted 90-day VTE rate in the aspirin group may have been due to selective use in lower-risk patients. This study was registered at ClinicalTrials.gov, trial number NCT01899443 (15/07/2013).


A case controlled study of serial transabdominal amnioinfusions in the management of second trimester oligohydramnios due to premature rupture of membranes.
The aim of this study was to evaluate the role of transabdominal amnioinfusion in relieving oligohydramnios and improving pregnancy outcome. Pregnant women with oligohydramnios amniotic fluid index (AFI<5) and premature rupture of membranes (PROM) from <27 weeks gestation were managed with serial transabdominal amnioinfusions. Under ultrasonic guidance, a 20-gauge needle was instilled in the uterine cavity and normal saline was infused until the AFI was normal. Repeat amnioinfusion was done weekly if oligohydramnios recurred. Amnioinfused cases were compared to cases with pPROM and oligohydramnios who had standard management. The mean gestational age at first procedure was 22 weeks. The mean pre-procedure AFI was 1.1cm and post-procedure was 12 cm. The mean number of infusions was 2.4. The mean first infusion to delivery interval was 33 days. The amnioinfused group when compared to the control group had decreased perinatal mortality of 33% versus 83% (P=0.036, OR=0.4, 95% CI=0.17-0.93), neonatal mortality of 17% versus 71% (P=0.049, OR=0.26, 95% CI=0.07-0.97) and neonatal sepsis 86% versus 27%(P=0.049, OR=0.32, 95% CI=0.12-0.87) with no statistical difference in gestational age at rupture and delivery nor birthweight. Babies discharged home compared to non-survivors had a significant increase in gestational age at delivery, birthweight, NICU days and transabdominal amnioinfusions. Logistic regression showed that only transabdominal amnioinfusion and gestational age correlated with survival. In selected cases of oligohydramnios with pPROM, transabdominal amnioinfusions may be associated with fluid retention and improved neonatal survival.


Novel synthesis of polymorphous nanocrystalline KNbO3 by a low temperature solution method.
Orthorhombic and rhombohedral single nanocrystalline KNbO3 with different morphologies was synthesized respectively at a relatively low temperature by simply changing the reaction solvent; a conceivable reaction process was illustrated based on the finding of a hexahedral intermediate product during the formation of KNbO3.


Randomized phase II study of gemcitabine and S-1 combination versus gemcitabine alone in the treatment of unresectable advanced pancreatic cancer (Japan Clinical Cancer Research Organization PC-01 study).
To evaluate the efficacy and safety of the combination of gemcitabine (GEM) and S-1 (GS) in comparison to GEM alone (G) for unresectable pancreatic cancer. In this multicenter randomized phase II study, we randomly assigned unresectable pancreatic cancer patients to either the GS group or the G group. The GS group regimen consists of intravenous 1,000 mg/m(2) GEM during 30 min on days 1 and 8, combined with 80 mg/m(2) oral S-1 twice daily on days 1-14, repeated every 3 weeks. On the other hand, the G group regimen consists of intravenous 1,000 mg/m(2) GEM on days 1, 8, and 15, repeated every 4 weeks. The primary endpoint was objective response rate (ORR). Secondary end points included treatment toxicity, clinical response benefit, progression-free survival (PFS), and overall survival. We registered 117 patients from 16 institutions between June 2007 and August, 2010. The ORR of the GS group was 28.3%, whereas that of the G group was 6.8%. This difference was statistically significant (P = 0.005). The disease control rate was 64.2% in the GS group and 44.1% in the G group. Median PFS was 6.15 months in the GS group and 3.78 month in the G group. This was also statistically significant (P = 0.0007). Moreover, the median overall survival (OS) of the GS group was significantly longer than that of the G group (13.7 months vs. 8.0 months; P = 0.035). The major grade 3-4 adverse events were neutropenia (54.7% in the GS group and 22.0% in the G group), thrombocytopenia (15.1% in the GS group and 5.1% in the G group), and skin rash (9.4% in the GS group). The GS group showed stronger anticancer activity than the G group, suggesting the need for a large randomized phase III study to confirm GS advantages in a specific subset.


Relationship between the dose-response effects of diazepam and clobazam on electroencephalographic parameters and on kindled amygdaloid seizure activity in rats.
1. The possibility that the anticonvulsant activity of the benzodiazepines, diazepam and clobazam, is related to changes in EEG parameters, particularly beta activity, was investigated in amygdaloid kindled rats. 2. The effects of diazepam (1, 2, 4, 8 and 16 mumols/kg), administered intraperitoneally (i.p.), clobazam (1, 2, 4, 8, 16 and 32 mumols/kg, i.p.) or vehicle (dimethyl sulfoxide) on the cortical EEG of amygdaloid kindled rats were quantitated for 15 min using computerized period amplitude analysis. Immediately afterwards, the amygdala was stimulated and the after-discharge duration (AD) and the seizures stage (SS) were determined. 3. The equivalent percentage time (EPT) of the diazepam-treated group was decreased in the theta band (4-8 Hz) and increased in the alpha (8-12 Hz) and first six beta (12-36 Hz) bands. The mean peak amplitude (MPA) was increased in the alpha (8-12 Hz) and all seven beta bands (12-40 Hz). Clobazam increased the EPT and MPA in the alpha (8-12 Hz) and all seven beta (12-40 Hz) bands. The MPA was also increased by clobazam in the theta (4-8 Hz) band. 4. Diazepam reduced both the AD and SS of the kindled seizures at doses of 4, 8 and 16 mumols/kg, whereas clobazam was anticonvulsant at doses of 16 and 32 mumols/kg. The reduction in both AD and SS correlated with increases in the EPT and MPA in the first beta (12-16 Hz) band in the diazepam-treated group and in the first four beta (12-28 Hz) bands in the clobazam-treated group.


[Obesity, a multifactorial, chronic disease].
A pathology of modernity, obesity is developing rapidly in the population, linked to multiple risk factors and somatopsychic disturbances. The healthcare system's responses have been enriched by specialized programs and centers, but we mustn't forget basic clinical pragmatism.


Biomatrix of health risk assessment of benzene-exposed workers at Thai gasoline stations.
This study assessed the health risk of benzene exposure among Thai gasoline station workers through biomarker detection and experience of adverse symptoms. Trans, trans-muconic acid (tt-MA) metabolites of benzene were analyzed from spot urine sampled among gasoline station workers after shift work using HPLC-UV. Air benzene monitoring was done with an active sampler connected to a charcoal sorbent tube, and analyzed by GC-FID. The health risk was calculated by using the biomatrix of the likelihood of benzene exposure and the severity of adverse symptoms. The tt-MA concentration, among 235 workers, ranged from less than 10-2159 µg/g Cr, which corresponded to the air benzene concentration range of <0.1 to 65.8 ppb. In total, 32.3% of workers had a higher than acceptable risk level and there was a significant association between gasoline station work zones and the likelihood of benzene exposure as well as the health risk of workers. The health risk levels estimated from the biomarker monitoring were consistent with the risk matrix of air benzene monitoring. This tt-MA biomarker monitoring and biomatrix of health risk assessment is suggested as useful for health surveillance of gasoline station workers exposed to benzene.


The slope of the segmental stretch-strain relationship as a noninvasive index of LV inotropy.
The aim of this study was to test the hypothesis that the noninvasively constructed slope of the relationship between left ventricular (LV) regional systolic strain and stretch during atrial contraction represents LV inotropic state. LV systolic response to a changing preload depends on its inotropic state. Changing the preload has allowed constructing the slope of the end-systolic pressure-volume relationship that is used as an invasive measurement of LV inotropy. We assumed that the slope of the relationship between regional systolic LV strain (total_S) and stretch during atrial contraction (preS) depends on the LV inotropic state as well and can thus be used as a LV inotropy index. Strain curves (tissue Doppler) were extracted from 27 healthy individuals to determine the normal stretch-strain relationship at rest, during a low-dose dobutamine (LD) challenge and during passive leg-lift (LL). The method was also applied in 7 patients with breast cancer before and after chemotherapy with anthracyclines. PreS and total_S correlated closely in all subjects (r = 0.82). Total_S values increased (p < 0.05) with LD (-20.44 ± 3.89% vs. -24.24 ± 5.55%) and LL (-19.65 ± 3.77% vs. -24.05 ± 3.67%), whereas preS increased only with LL (5.96 ± 1.72% vs. 8.61 ± 2.18%), but not with LD (6.83 ± 2.34% vs. 7.29 ± 2.24%). No changes of total_S or preS were observed after the exposure to chemotherapy (-21.23 ± 2.93% vs. -21.49 ± 2.89% and 8.11 ± 1.03% vs. 8.59 ± 1.73%, respectively). The slope of stretch-strain relationship got steeper with LD (-1.47 ± 0.36 vs. -2.34 ± 0.36, p < 0.05), declined after the chemotherapy (-1.68 ± 0.15 to -0.86 ± 0.23, p < 0.05) and did not change with LL (-1.39 ± 0.57 vs. -1.51 ± 0.38, p = NS). The slope of the regional stretch-strain relationship can be regarded as a noninvasive index of myocardial inotropic state. It gets steeper with increasing inotropy, does not change with preload induced changes of LV systolic function, and flattens after the exposure to a cardiotoxic drug.


Measurement of urinary N-acetyl-b-D-glucosaminidase in adult patients with cystic fibrosis: before, during and after treatment with intravenous antibiotics.
Patients with cystic fibrosis (CF) are at high risk from the nephrotoxic effects of intravenous antibiotics due to repeated and prolonged courses of therapy. Routine methods of monitoring renal injury are insensitive. N-acetyl-b-d-glucosaminidase (NAG) is a lysosomal enzyme present in the renal proximal tubular cells, with increased excretion an indicator of renal tubular dysfunction. Urinary NAG, creatinine, serum creatinine, electrolytes and BUN were measured on days 1, 14 and at the first out-patient visit following treatment with tobramycin or colistin. Urinary NAG levels were corrected for urinary creatinine and expressed as a NAG ratio. Patients who received>1 course of intravenous antibiotics during the study period were included in a separate analysis of the cumulative effect of treatment. 88 patients (44 female, 31 with CFRD) completed a single course of intravenous antibiotics. 71 patients had urinary NAG levels at follow-up. The median time to follow-up was 50 days. Serum electrolytes, creatinine and BUN were normal throughout. A 3.5-fold increase in urinary NAG excretion was observed between day 1 and 14 and 46% of patients had an elevated NAG level at follow-up. A highly significant difference in NAG excretion was observed on day 14 for tobramycin vs. colistin (median 2.24 vs. 0.98, p<0.001). A significant difference in NAG excretion was seen in patients with CFRD at all measured time points. Patients with CFRD had a significantly worse clinical status and had received more days of intravenous antibiotics over the previous 6 years. In 20 (80%) of 25 patients who received>1 course of treatment during the study period, baseline NAG levels were significantly higher in subsequent courses (p<0.001). There was a significant correlation between previous exposure to colistin and baseline NAG levels (r=0.389, p<0.001). Both tobramycin and colistin cause acute renal tubular injury with a significant rise in urinary NAG excretion. Patients with CFRD seem to be at greatest risk of renal tubular damage. Cumulative damage is evident with repeated dosing. Previous exposure to nephrotoxic antibiotics, especially colistin, is associated with elevated baseline NAG levels. We recommend that colistin is reserved for patients with resistant Pseudomonas aeruginosa or those who are intolerant to tobramycin. Serial longitudinal NAG measurements may be useful in patients with CF, especially those with CFRD, to identify patients at risk of developing renal disease.


Cell migration strategies in 3-D extracellular matrix: differences in morphology, cell matrix interactions, and integrin function.
Cell migration in extracellular matrix is a complex process of adhesion and deadhesion events combined with cellular strategies to overcome the biophysical resistance imposed by three-dimensionally interconnected matrix ligands. Using a 3-D collagen matrix migration model in combination with computer-assisted cell tracking for reconstruction of migration paths and confocal microscopy, we investigated molecular principles governing cell-matrix interactions and migration of different cell types. Highly invasive MV3 melanoma cells and fibroblasts are large and highly polarized cells migrating at low speed (0.1-0.5 microm/min) and at high directional persistence. MV3 melanoma cells utilize adhesive migration strategies as characterized by high beta1 integrin surface expression, beta1 integrin clustering at interactions with matrix fibers, and beta1 integrin-mediated adhesion for force generation and migration. In contrast, T lymphocytes and dendritic cells are highly mobile cells of lower beta1 integrin expression migrating at 10- to 40-fold higher velocities, and directionally unpredictable path profiles. This migration occurs in the absence of focal adhesions and largely independent of beta1 integrin-mediated adhesion. Whereas cell-matrix interactions of migrating tumor cells result in traction and reorientation of collagen fibers, partial matrix degradation, and pore formation, leukocytes form transient and short-lived interactions with the collagen lacking structural proteolysis and matrix remodeling. In conclusion, the 3-D extracellular matrix provides a spatially complex and biomechanically demanding substrate for cell migration, thereby differing from cell migration across planar ligands. Highly adhesive and integrin-dependent migration strategies detected in morphologically large and slowly migrating cells may result in reorganization of the extracellular matrix, whereas leukocytes favor largely integrin-independent, rapid, and flexible migration strategies lacking typical focal adhesions and structural matrix remodeling.


Dynamic phenotypic restructuring of the CD4 and CD8 T-cell subsets with age in healthy humans: a compartmental model analysis.
In healthy humans, phenotypic restructuring occurs with age within the CD3+ T-lymphocyte complement. This is characterized by a non-linear decrease of the percentage of 'naive' (CD45RA+) cells and a corresponding non-linear increase of the percentage of 'memory' (CD45R0+) cells among both the CD4+ and CD8+ T-cell subsets. We devised a simple compartmental model to study the age-dependent kinetics of phenotypic restructuring. We also derived differential equations whose parameters determined yearly gains minus losses of the percentage and absolute numbers of circulating naive cells, yearly gains minus losses of the percentage and absolute numbers of circulating memory cells, and the yearly rate of conversion of naive to memory cells. Solutions of these evaluative differential equations demonstrate the following: (1) the memory cell complement 'resides' within its compartment for a longer time than the naive cell complement within its compartment for both CD4 and CD8 cells; (2) the average, annual 'turnover rate' is the same for CD4 and CD8 naive cells. In contrast, the average, annual 'turnover rate' for memory CD8 cells is 1.5 times that of memory CD4 cells; (3) the average, annual conversion rate of CD4 naive cells to memory cells is twice that of the CD8 conversion rate; (4) a transition in dynamic restructuring occurs during the third decade of life that is due to these differences in turnover and conversion rates, between and from naive to memory cells.


Monoclonal antibody analysis of mononuclear cells in myopathies. IV: Cell-mediated cytotoxicity and muscle fiber necrosis.
Cell-mediated muscle fiber injury occurs in inclusion body myositis (IBM), polymyositis (PM), and even in Duchenne dystrophy (DD). Most of the autoaggressive cells are T cells and macrophages, but some are killer/natural killer (K/NK) cells. We here compare the frequencies per 1,000 muscle fibers of endomysial K/NK cells of varying cytotoxicity with those of T cells and macrophages in 8 cases each of IBM, PM, and DD. Two-micrometer serial cryostat sections were analyzed. The Leu-4 marker, present on all T cells and some K/NK cells, and the Leu-7 and Leu-11 markers, present on K/NK cells, were localized by paired immunofluorescence. Macrophages were demonstrated by the acid phosphatase reaction. In IBM, PM and DD, the respective average cell counts per 1,000 muscle fibers were: Leu-4+7- cells (T cells not expressing a K/NK marker)--710, 530, and 59; Leu-4+7+ cells (K/NK cells of low K/NK cytotoxicity)--294, 163, and 13; Leu-4-7+ cells (K/NK cells of intermediate cytotoxicity)--32, 10, and 2; and macrophages--292, 251, and 38. Leu-11+ K/NK cells that have the highest killing activity were virtually absent in all cases. The data suggest a limited role for antigen and major histocompatibility complex unrestricted K/NK cells, as compared with antigen-specific and major histocompatibility complex-restricted T cells, in IBM and PM. Further, the findings cast doubt on the significance of either T cells or K/NK cells in mature muscle in DD.(ABSTRACT TRUNCATED AT 250 WORDS)


Hemodynamic responses of chronically instrumented piglets to bolus injections of group B streptococci.
Group B beta-hemolytic Streptotocci cause pulmonary hypertension when injected into animals and may precipitate the persistent pulmonary hypertension syndrome in infected human neonates. We used chronically instrumented piglets to study the effects of repeated injections of heat-killed group B Streptococcus (GBS) type III. Daily exposure to GBS was associated with a 2-fold or greater potentiation of pulmonary and systemic hypertensive responses after 1 wk. Throughout experimentation, pulmonary pressure changes were more marked than systemic changes. After establishing a dose-response relationship, we chose a control dose that produced intermediate hypertensive responses. We then evaluated the effects of antibody and various drugs on the hypertensive responses. Preincubation of organisms with rabbit antiserum containing type-specific antibody enhanced the responses. Beta endorphin blockade with naloxone had little or no effect; leukotriene synthesis inhibition also did not affect responses. Both indomethacin, a cyclooxygenase inhibitor, and dazmegrel, a specific thromboxane synthesis inhibitor, blocked the hypertensive responses to GBS. It appears that repeated doses of GBS potentiate the hypertensive responses, a process that we hypothesize may be mediated by development of type-specific antibody as type-specific antibody levels rose during potentiation. It is likely that thromboxane A2 is the effector of the pulmonary and systemic hypertensive responses to GBS injection, because thromboxane inhibition by dazmegrel was as effective as indomethacin in blocking these effects. Thromboxane synthesis blockade may prove useful in management of hemodynamic disturbances accompanying severe bacterial infections in humans.


Humoral and cellular rejection after combined liver-kidney transplantation in low immunologic risk recipients.
Combined liver-kidney transplantation is considered a low risk for immunologic complication. We report an unusual case of identical ABO liver-kidney recipient without preformed anti-human leukocyte antigen (HLA) antibodies, transplanted across a T- and B-cell-negative cross-match and complicated by early acute humoral and cellular rejection, first in the liver then in the kidney. While analyzing the immunologic complications in our cohort of 12 low-risk combined liver-kidney recipients, only one recipient experienced a rejection episode without detection of anti-HLA antibody over time. Although humoral or cellular rejection is rare after combined kidney-liver transplantation, our data suggest that even in low-risk recipients, the liver does not always systematically protect the kidney from acute rejection. Indeed, the detection of C4d in the liver should be carefully followed after combined liver-kidney transplantation.


Effect of chitosan on epithelial cell tight junctions.
Chitosan has been proposed as a novel excipient for transepithelial drug-delivery systems. Chitosan is thought to disrupt intercellular tight junctions, thus increasing the permeability of an epithelium. The effect of chitosan on tight junction complex was investigated at the molecular level. Changes in barrier properties of Caco-2 cell monolayers, including transepithelial electrical resistance and permeability to horseradish peroxidase (HRP), were assessed in response to chitosan treatment. Changes in subcellular localization of the tight junction proteins zona occludens 1 (ZO-1) and occludin by immunofluorescence and Western blotting of cellular fractions were also assessed. Chitosan was found to cause a dose-dependent reduction in transepithelial electrical resistance of Caco-2 monolayers of up to 83%. A corresponding increase in horseradish peroxidase permeability of up to 18 times greater than the control was also observed across the monolayer. Immunofluorescent localization of ZO-1 revealed loss of membrane-associated ZO-1 from discrete areas. Analysis of cellular fractions revealed a dose-dependent loss of ZO-1 and occludin from the cytosolic and membrane fractions into the cytoskeletal fraction. These changes did not occur because of chitosan-mediated ATP depletion. Chitosan-mediated tight junction disruption is caused by a translocation of tight junction proteins from the membrane to the cytoskeleton.