1H-HRMAS NMR study of cold smoked Atlantic salmon (Salmo salar) treated with E-beam.
(1)H High resolution magic angle spinning (HRMAS) NMR spectroscopic data in combination with principal components analysis and analysis of variance were used to differentiate between irradiated and non-irradiated cold-smoked Atlantic salmon (Salmo salar). NMR profiling was obtained, with a spectral acquisition time of less than 8 min, from a small sample size of intact white salmon muscle, by non-destructive analysis that includes a very simple and rapid sample preparation step. Results obtained enable the use of creatine, trimethylamine oxide and the sum of phosphorylcholine and glycerophosphorylcholine as diagnostic compounds to detect irradiation treatment. This study shows the potential of (1)H-HRMAS to be a rapid method for investigating compositional changes due to food processing as well as to confirm the presence or absence of some bioactive compounds in irradiated samples.


[Experience of first COVID-19 patients in Northern part of the Netherlands: Information provision, source investigation, contact tracing and home isolation].
The aim of this study was to investigate how patients experience the information, the source investigation and contact tracing and the measures in isolation at the start of a pandemic. Secondary analysis of semi-structured interviews was conducted as part of a larger exploratory mixed-methods study on COVID-19 patient experiences. Semi-structured interviews were conducted with 29 people from Friesland and Groningen who tested positive for SARS-CoV-2 between 9 March and 3 April 2020, recruited via maximum variation sampling. Thematic analysis was used. The following themes emerged from the analysis: 1) Information: Conflicting information by different advisors led to a lack of clarity. Patients admitted to hospital usually felt uninformed about the rules of home isolation after discharge. 2) Investigation into the source of infection: For most it was unclear whether and how this investigation took place. Some expected feedback on their suggestions. 3) Informing contacts: Not everyone felt able to inform the right contacts. Some felt stigmatized. 4) Living with the measures in home isolation: The recommended living rules were often not fully applied. Some patients felt insufficiently supported. Our study shows that not all COVID-19 patients felt well cared for at the start of the outbreak. Scaling down monitoring by the public health service can mean that questions about source and contact investigation and isolation remain unanswered or are answered by others. This leads to conflicting information and non-compliance with measures. The supervision of patients in isolation should be better guarded.


High-dose chemo-radiotherapy followed by autologous Philadelphia chromosome-negative blood progenitor cell transplantation in patients with chronic myelogenous leukemia.
Twenty-three patients with chronic myelogenous leukemia in early chronic phase (ECP) and not previously treated with alpha-interferon (IFN-alpha) (10 patients), in ECP but pretreated with IFN-alpha (<12 months) (seven patients) and in late chronic phase (LCP) pretreated with IFN-alpha (>12 months) (six patients) underwent autografting with Philadelphia (Ph) chromosome-negative blood progenitor cells (BPCs) (20 patients), or partially/totally Ph-positive BPCs (three patients), previously mobilized during the early phase of recovery after aplasia induced by intensive chemotherapy. The conditioning regimen consisted of high-dose chemotherapy alone or followed by total body irradiation (TBI). Recombinant G-CSF was given after BPCs infusion on day +8. All patients in ECP not pretreated with IFN-alpha are alive and five of them are Ph-negative in the marrow after autografting. Six of seven patients autografted with Ph-negative BPCs in the group of ECP pretreated with IFN-alpha (<12 months) are alive and two of them are still Ph-negative in the marrow. In the same group, the only patient transplanted with partially Ph-positive BPCs, died of blastic transformation 2 months after reinfusion. Three patients (two patients autografted with Ph-negative BPCs and one patient with Ph-positive BPC) in the group of LCP pretreated with IFN-alpha >12 months are alive but Ph-positive after autografting. The other three patients of the same group died of procedure-related toxicity (two patients) and blastic transformation (one patient). Seventeen patients (10/10 ECP not pretreated with IFN-alpha; 5/7 ECP pretreated with IFN-alpha and 2/6 LCP pretreated with IFN-alpha) of 23 autografted patients were treated with IFN-alpha +/- IL-2. Toxicities after autografting were mostly related to myelosuppression, particularly thrombocytopenia. All patients of the two groups pretreated with IFN-alpha developed febrile episodes during the aplastic phase following BPCs reinfusion. No patient autografted in ECP and those not pretreated with IFN-alpha developed febrile episodes. This is also probably due to the use of i.v. antibiotic and antimicotic prophylaxis when neutrophils were < or = 1 x 10(9)/l after autografting. Greater toxicity was observed in patients pretreated with IFN-alpha, being lethal in two cases in LCF. In conclusion, the "in vivo' manipulation approach employed in our institution is a safe procedure and it results in a high collection of Ph-negative cells in the blood if the cells are harvested: (1) in early chronic phase; (2) in early phase of recovery after chemotherapy-inducing aplasia; (3) in patients not extensively pretreated with IFN-alpha. The data presented here have shown encouraging trends in chronic phase of CML and offer new perspective for patients without an HLA-identical donor or for patients who do not respond to IFN-alpha.


[Suppression of lipid peroxidation by sho-saiko-to and its components in rat liver subcellular membranes].
In order to elucidate anti-inflammatory action of Sho-saiko-to, its components were analyzed and purified by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), and their effects on lipid peroxidation were compared with those of Sho-saiko-to in guinea pig neutrophils, rat liver mitochondria and microsomes. The results obtained were as follows: 1) Sho-saiko-to gave no effect on arachidonate- and PMA (phorbol 13-myristate 12-acetate)-induced NADPH-O2- generation in neutrophils though it gave a weak LPS-like effect on the cells. No inhibition of reduced nicotinamido adenine dinucleotide phosphate (NADPH) oxidation was shown in liver microsomes. However, it markedly inhibited iron-induced lipid peroxidation in microsomes and mitochondria. 2) The active components to suppress lipid peroxidation could be concentrated in a lipid-soluble fraction from the Sho-saiko-to solution. The ED50 values to give 50% inhibition of the mitochondrial lipid peroxidation were 160 ng/mg prot, and 50 micrograms/mg prot, for the lipid-soluble fraction and Sho-saiko-to, respectively. 3) The components in the lipid-soluble fraction were further separated by TLC and HPLC, and their inhibitory effects on the lipid peroxidation were examined. The active components were baicalein, ginsenoside Rf and baicalin, of which ED50 values were 0.2, 0.2 and 1 nmol/mg prot, respectively. Glycyrrhizin and its derivatives, ginsenoside (except ginsenoside Rf), and saikosaponins gave no effect in the concentration examined. 4) From these results anti-inflammatory action of Sho-saiko-to was discussed.


CD8dimCD3+ lymphocytes in fever patients might be biomarkers of active EBV infection and exclusion indicator of T-LGLL.
Background: Massive monoclonal or oligoclonal expansion of CD8+ T cells is a notable feature of primary infections of the Epstein-Barr virus (EBV). However, the clinical significance of this expansion is not clear. Results: An increase in the CD8dimCD3+ lymphocyte subset in patients with active EBV infection was due to caspase-8-dependent apoptosis was found using flow cytometry in this study. The number of these cells was associated with the illness severity. Pan-T-cell antigen and receptor analyses were also compared in patients with active EBV infections and T-cell large granular lymphocytic leukemia to provide additional diagnostic information. Conclusion: The increase in CD8dimCD3+ cells could be a biomarker of active EBV infection and an exclusion indicator of T-cell large granular lymphocytic leukemia with flow cytometric analysis.


MicroRNA Profiling of the Effect of the Heptapeptide Angiotensin-(1-7) in A549 Lung Tumor Cells Reveals a Role for miRNA149-3p in Cellular Migration Processes.
Lung cancer is one of the most frequent types of cancer in humans and a leading cause of death worldwide. The high mortality rates are correlated with late diagnosis, which leads to high rates of metastasis found in patients. Thus, despite all the improvement in therapeutic approaches, the development of new drugs that control cancer cell migration and metastasis are required. The heptapeptide angiotensin-(1-7) [ang-(1-7)] has demonstrated the ability to control the growth rates of human lung cancer cells in vitro and in vivo, and the elucidation of central elements that control the fine-tuning of cancer cells migration in the presence of the ang-(1-7), will support the development of new therapeutic approaches. Ang-(1-7) is a peptide hormone of the renin-angiotensin system (RAS) and this study investigates the modulatory effect of the heptapeptide on the expression pattern of microRNAs (miRNAs) in lung tumor cells, to elucidate mechanistic concerns about the effect of the peptide in the control of tumor migratory processes. Our primary aim was to compare the miRNA profiling between treated and untreated-heptapeptide cells to characterize the relevant molecule that modulates cellular migration rates. The analyses selected twenty one miRNAs, which are differentially expressed between the groups; however, statistical analyses indicated miRNA-149-3p as a relevant molecule. Once functional analyses were performed, we demonstrated that miRNA-149-3p plays a role in the cellular migration processes. This information could be useful for future investigations on drug development.


Physiological impairment in linoleic acid deficiency of rats and the effect of n-3 polyunsaturated fatty acids.
Some impairments related to membrane function were found in linoleic acid-deficient rats and the effects of fish oil feeding were investigated. In linoleic acid-deficient rats, glucose transport into erythrocytes was decreased. The concentrations of plasma free fatty acids were significantly reduced in the animals. Further, epinephrine-stimulated lipase was remarkably less sensitive to epinephrine in the deficient rat than in the corn oil-fed control rat. However, these impairments were relieved by fish oil feeding. Therefore, the impairments may be ascribed to the decrease of arachidonic acid as a polyunsaturated fatty acid in membrane phospholipids, since n-3 polyunsaturated fatty acids appear to take the place of arachidonic acid.


A de novo loss-of-function GRIN2A mutation associated with childhood focal epilepsy and acquired epileptic aphasia.
N-methyl-D-aspartate receptors (NMDAR) subunit GRIN2A/GluN2A mutations have been identified in patients with various neurological diseases, such as epilepsy and intellectual disability / developmental delay (ID/DD). In this study, we investigated the phenotype and underlying molecular mechanism of a GRIN2A missense mutation identified by next generation sequencing on idiopathic focal epilepsy using in vitro electrophysiology. Genomic DNA of patients with epilepsy and ID/DD were sequenced by targeted next-generation sequencing within 300 genes related to epilepsy and ID/DD. The effects of one missense GRIN2A mutation on NMDAR function were evaluated by two-electrode voltage clamp current recordings and whole cell voltage clamp current recordings. We identified one de novo missense GRIN2A mutation (Asp731Asn, GluN2A(D731N)) in a child with unexplained epilepsy and DD. The D731N mutation is located in a portion of the agonist-binding domain (ABD) in the GluN2A subunit, which is the binding pocket for agonist glutamate. This residue in the ABD is conserved among vertebrate species and all other NMDAR subunits, suggesting an important role in receptor function. The proband shows developmental delay as well as EEG-confirmed seizure activity. Functional analyses reveal that the GluN2A(D731N) mutation decreases glutamate potency by over 3,000-fold, reduces amplitude of current response, shortens synaptic-like response time course, and decreases channel open probability, while enhancing sensitivity to negative allosteric modulators, including extracellular proton and zinc inhibition. The combined effects reduce NMDAR function. We identified a de novo missense mutation in the GRIN2A gene in a patient with childhood focal epilepsy and acquired epileptic aphasia. The mutant decreases NMDAR activation suggesting NMDAR hypofunction may contribute to the epilepsy pathogenesis.


Acid-Base and Plasma Biochemical Changes Using Crystalloid Fluids in Stranded Juvenile Loggerhead Sea Turtles (Caretta caretta).
The aim of this study was to compare the efficacy and effects on acid-base and electrolyte status of several crystalloid fluids in 57 stranded juvenile loggerhead turtles. Within a rehabilitation program four different crystalloid fluids were administered (0.9% Na Cl solution; 5% dextrose + 0.9% Na Cl solutions 1:1; 0.9% Na Cl + lactated Ringer's solutions 1:1; lactated Ringer's solution). Crystalloid fluids were intracoelomically administered during three days (20 ml/kg/day). Animals were sampled at three different moments: Upon admission for evaluating the type of acid-base or biochemical disorder, post-fluid therapy treatment for controlling the evolution of the disorder, and post-recovery period for obtaining the baseline values for rehabilitated loggerhead turtles. Each sample was analyzed with a portable electronic blood analyzer for pH, pO2, pCO2, lactate, sodium, potassium, chloride, glucose, and BUN concentration. Admission and post-fluid therapy treatment values were compared with those obtained for each turtle immediately before release. The highest percentage of acid-base recovery and electrolyte balance was observed in turtles treated with mixed saline-lactated Ringer's solution (63.6%), followed by turtles treated with physiological saline solution (55%), lactated Ringer's solution (33.3%), and dextrose-saline solutions (10%). Most turtles treated with lactated Ringer's solution had lower lactate concentrations compared with their initial values; however, 66.6% of turtles treated with lactated Ringer's solution had metabolic alkalosis after therapy. Significant higher concentrations of glucose were detected after saline-dextrose administration compared with all the remaining fluids. This is the first study evaluating the effects of several crystalloid fluids on the acid-base status and plasma biochemical values in stranded loggerhead sea turtles. Reference convalescent venous blood gas, acid-base, and plasma biochemical values, useful for veterinary surgeons involved in sea turtle conservation, are also provided.


Definitive 3D-CRT for clinically localized prostate cancer: modifications of the clinical target volume following a prostate MRI and the clinical benefits.
To evaluate the modifications of the tumor stage and clinical target volume following a prostate magnetic resonance imaging (MRI) to evaluate the tumor (T) staging, and the clinical benefits for prostate cancer. A total of 410 patients with newly diagnosed and clinically localized prostate cancer were retrospectively analyzed. The patients were treated with definitive three-dimensional conformal radiotherapy (3D-CRT). In all of the patients, digital rectal examination, transrectal ultrasound, prostate biopsy and computed tomography were performed to evaluate the clinical stage. Of the 410 patients, 189 patients had undergone a prostate MRI study to evaluate the T staging, and 221 patients had not. Modification of the T stage after the prostate MRI was seen in 39 (25%) of the 157 evaluable patients, and a modification of the risk group was made in 14 (9%) patients. Eventually, a modification of the CTV in 3D-CRT planning was made in 13 (8%) patients, and 10 of these had extracapsular disease. Most of the other modifications of the T staging were associated with intracapsular lesions of prostate cancer which did not change the CTV. There were no significant differences in the biological relapse-free survival between the patients with and without a prostate MRI study. Modification of the CTV were recognized in only 8% of the patients, most of whom had extracapsular disease, although that of the T stage was seen in approximately one-quarter of the patients. Prostate MRI should only be selected for patients with a high probability of extracapsular involvement.


Airway microbiota correlated with pulmonary exacerbation in primary ciliary dyskinesia patients.
PCD is a rare disease characterized by productive cough, rhinitis, and recurrent infections of the upper and lower airways. Because the diagnosis of PCD is often delayed, patients receive more antibiotics, experience a heavier financial burden, and have a worse prognosis; thus, it is very important to identify the pathogeny and use the correct antibiotic. In this large single-center study of PCD microbiota, we identified an outline of the bacterial microbes from the respiratory tract; furthermore, we found that the microbiota diversity in pediatric sputum was richer than that in pediatric BALF through sequencing, indicating a heterogeneous community structure. The microbiota diversity and richness were lower during pulmonary exacerbation than during pulmonary stabilization. A significantly higher abundance of Pseudomonas had a moderate distinguishing effect for lung exacerbation, which attracted more attention for the study of Pseudomonas therapy in pediatric patients with PCD.


The role of crumbs genes in the vertebrate cornea.
To evaluate the role of crumbs genes and related epithelial polarity loci in the vertebrate cornea. The authors used histologic analysis and electron microscopy to evaluate the corneas of zebrafish mutant for a crumbs locus oko meduzy (ome) and in mutants of four other loci, nagie oko (nok), heart and soul (has), mosaic eyes (moe), and ncad (formerly glass onion), that function in the same or related genetic pathways. In parallel, they performed an evaluation of corneas in human carriers of a crumbs gene, CRB1, and mutations using topography and biomicroscopy. The expression of the CRB1 gene in the normal human cornea was examined by polymerase chain reaction (PCR) and immunohistochemical staining. The corneas of zebrafish mutants display severe abnormalities of the epithelial and stromal layers. The epithelial cells do not properly adhere to each other, and fluid-filled spaces form between them. In addition, the layering of the corneal stroma is poorly formed or absent. The corneas of human carriers of CRB1 mutations display shape deviations compared with what has been observed in normal individuals. A PCR product of the correct size was obtained from normal human corneal samples. Sequence analyses confirmed its identity to be the human CRB1 gene. Immunohistochemical staining using anti-CRB1 yielded positive brown deposits in the human cornea. crumbs genes play a role in the differentiation of the vertebrate cornea. Corneal defects associated with crumbs gene mutations are very severe in the zebrafish model and, in comparison, appear clinically less pronounced in the human eye.


Estrogen receptors in giant cell arteritis. An immunocytochemical, western blot and RT-PCR study.
Giant cell arteritis (GCA) is a chronic form of vasculitis which predominantly affects women over 50 years of age. The aim of this study was to analyse the presence of estrogen receptor alpha (ER) in the temporal arteries of patients with GCA. Inflamed temporal artery biopsies from 43 GCA patients were stained with monoclonal antibodies to two different segments of the ER and compared with non-inflamed arteries from age- and sex-matched controls who had not received a clinical diagnosis of GCA. The protein that was extracted from 4 GCA-positive biopsies and 4 non-GCA controls was analysed using the Western blot method with a monoclonal antibody to ER. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis using primer pairs specific to ER-cDNA was performed on the total RNA from 4 GCA-positive biopsies and 4 non-GCA controls. The inflamed arteries expressed distinct cytoplasmic immunoreactivity to ER in activated mononuclear inflammatory cells and in giant cells. Biopsies from GCA patients and controls displayed cytoplasmic ER positivity in smooth muscle cells. Western blot analysis revealed two bands corresponding to approximately 64 and 54 kDa, respectively, in the inflamed arteries and controls. In the inflamed biopsies and non-GCA controls, RT-PCR analysis revealed a strong band corresponding to approximately 670 bp, as expected, and a weaker band corresponding to approximately 440 bp. In inflamed arteries from GCA patients, smooth muscle cells, activated mononuclear inflammatory cells and giant cells express cytoplasmic ER. Non-inflamed control arteries also express cytoplasmic ER in smooth muscle cells. The accumulation of cytoplasmic ER may suggest the involvement of estrogen not only in GCA but also in normal vascular aging. The results justify further investigations into the pathogenetic roles of estrogen metabolism in GCA.


An independent evaluation of four quantitative emergency department crowding scales.
Emergency department (ED) overcrowding has become a frequent topic of investigation. Despite a significant body of research, there is no standard definition or measurement of ED crowding. Four quantitative scales for ED crowding have been proposed in the literature: the Real-time Emergency Analysis of Demand Indicators (READI), the Emergency Department Work Index (EDWIN), the National Emergency Department Overcrowding Study (NEDOCS) scale, and the Emergency Department Crowding Scale (EDCS). These four scales have yet to be independently evaluated and compared. The goals of this study were to formally compare four existing quantitative ED crowding scales by measuring their ability to detect instances of perceived ED crowding and to determine whether any of these scales provide a generalizable solution for measuring ED crowding. Data were collected at two-hour intervals over 135 consecutive sampling instances. Physician and nurse agreement was assessed using weighted kappa statistics. The crowding scales were compared via correlation statistics and their ability to predict perceived instances of ED crowding. Sensitivity, specificity, and positive predictive values were calculated at site-specific cut points and at the recommended thresholds. All four of the crowding scales were significantly correlated, but their predictive abilities varied widely. NEDOCS had the highest area under the receiver operating characteristic curve (AROC) (0.92), while EDCS had the lowest (0.64). The recommended thresholds for the crowding scales were rarely exceeded; therefore, the scales were adjusted to site-specific cut points. At a site-specific cut point of 37.19, NEDOCS had the highest sensitivity (0.81), specificity (0.87), and positive predictive value (0.62). At the study site, the suggested thresholds of the published crowding scales did not agree with providers' perceptions of ED crowding. Even after adjusting the scales to site-specific thresholds, a relatively low prevalence of ED crowding resulted in unacceptably low positive predictive values for each scale. These results indicate that these crowding scales lack scalability and do not perform as designed in EDs where crowding is not the norm. However, two of the crowding scales, EDWIN and NEDOCS, and one of the READI subscales, bed ratio, yielded good predictive power (AROC >0.80) of perceived ED crowding, suggesting that they could be used effectively after a period of site-specific calibration at EDs where crowding is a frequent occurrence.


Antibacterial and antifungal properties of human cerumen.
To assess the antibacterial and antifungal properties of human cerumen by studying its effect on the growth of Staphylococcus aureus, Esherichia coli, Pseudomonas aeruginosa and Candida albicans. Cerumen samples were collected from 75 normal, healthy subjects aged from seven to 80 years, without ear pathology, who attended the ear, nose and throat out-patient clinic of the University Malaya Medical Center from May 2006 to October 2006. Of these 75 samples, 31 had no growth when cultured on nutrient agar. Inhibition studies on these 31 samples were performed for Staphylococcus aureus (American Type Culture Collection (ATCC) 25923), Esherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and Candida albicans. Nutrient agar was used to conserve all three bacterial strains and Sabouraud dextrose agar was used for Candida albicans. A decrease in Staphylococcus aureus growth was observed for 27 of the 31 samples. All 31 samples induced decreased growth of Pseudomonas aeruginosa, while 29 induced decreased growth of Candida albicans. However, only four samples induced decreased growth of Escherichia coli. Cerumen was demonstrated to have potential antimicrobial effects on strains of Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans.


Does the menstrual cycle influence the sensitivity of vagally mediated baroreflexes?
The menstrual cycle provokes several physiological changes that could influence autonomic regulatory mechanisms. We studied the carotid-cardiac baroreflex in ten healthy young women on four occasions over the course of their menstrual cycles (days 0-8, 9-14, 15-20 and 21-25). We drew blood during each session for analysis of oestrogen, progesterone and noradrenaline (norepinephrine) levels, and assessed carotid-cardiac baroreflex function by analysing R-R interval responses to graded neck pressure sequences. Oestrogen levels followed a classical two-peak (cubic) response, with elevated levels on days 9-14 and 21-25 compared with days 0-8 and 15-20 (P=0.0032), while progesterone levels increased exponentially from days 9-14 to days 21-25 (P=0.0063). Noradrenaline levels increased from an average of 137 pg/ml during the first three measurement periods to 199 pg/ml during days 21-25 (P=0.0456). Carotid-cardiac baroreflex gain and operational point were not statistically different at any of the time points during the menstrual cycle (P> or =0.18). These findings are consistent with the notion that beat-to-beat vagal-cardiac regulation does not change over the course of the normal menstrual cycle.


Estimation of drug-metabolizing capacity by cytochrome P450 genotyping and expression.
Many undesired side effects or therapeutic failures of drugs are the result of differences or changes in drug metabolism, primarily depending on the levels and activities of cytochrome P450 (P450) enzymes. To assess whether P450 expression profiles can reflect hepatic drug metabolism, we compared P450 mRNA levels in the liver or peripheral leukocytes with the corresponding hepatic P450 activities. A preliminary P450 genotyping for the most frequent polymorphisms in white populations (CYP2C9*2, CYP2C9*3, CYP2C19*2, CYP2C19*3, CYP2D6*3, CYP2D6*4, CYP2D6*6, and CYP3A5*3) was carried out before P450 phenotyping, excluding the donors with nonfunctional alleles of CYP2C9, CYP2C19, and CYP2D6 and those with a functional CYP3A5*1 allele from a correlation analysis. The hepatic mRNA levels of CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 displayed a strong association with P450 activities in the liver, whereas the expression of CYP1A2, CYP2C9, CYP2C19, and CYP3A4 in leukocytes was proven to reflect the hepatic activities of these P450 species. The leukocytes were found to be inappropriate cells for the assessment of hepatic CYP2B6 and CYP2D6 activities. Combining the results of P450 genotyping and phenotyping analyses, patients' drug-metabolizing capacities can be estimated by the P450 expression in the liver and in leukocytes with some limitations. Patients' genetic and nongenetic variations in P450 status can guide the appropriate selection of drugs and the optimal dose, minimizing the risk of harmful side effects and ensuring a successful outcome of drug therapy.


[Needle laparoscopic varicocelectomy].
We evaluated our method and effects of needle laparoscopic varicocelectomy for the treatment of varicocele. 72 patients (105 lateral) diagnosed varicocele were performed laparoscopic varicocelectomy under epidural combined intravenous anesthesia from Feb, 2003 to Apr, 2005. Two 2 mm incisions and one 5 mm incision were made on the midline of lower abdomen, by which two 2 mm trocars and one 5 mm trocar were introduced. Vessel-sealing device (Ligasure) was used to seal the internal spermatic veins. All operations were completed successfully. Mean operation time was 15 minutes unilateral and 21 minutes bilateral. The patients were hospitalized for 3 to 5 days after procedure. Follow-up was scheduled for 6 to 12 months and there was no recurrence. Needle laparoscopic varicocelectomy gives favorable effect with minimal invasion, rapid recovery, which is the best choice for the treatment of varicocele.


Ultrastructural pathological features of unilateral renal artery stenosis in the rats.
Renal artery stenosis (RAS) is one of the main reasons of renovascular hypertension and its pathogenesis remains unclear. In this study, we aimed to investigate histopathological characteristics in a rat model of RAS. Sprague-Dawley (SD) male rats were randomly divided into unilateral RAS group (Model group, n=30) and Sham group (n=30). The left renal artery was clamped with miniature silver clip for the rats in RAS group, while it was exposed but not clamped for the rats in Sham group. After the surgery, the rats were randomly divided into ten subgroups based on the time after surgery (n=3). Blood pressure, urinary albumin/creatinine ratio, and serum albumin and creatinine levels were measured. The kidneys were dissected for histological and electron microscopy analysis. The results showed that systolic blood pressure was significantly higher since 4 weeks after surgery compared to before surgery. There were no significant differences in urinary albumin/creatinine ratio as well as serum albumin and creatinine levels in Model and Sham groups. During the early acute renal ischemia the stenotic kidney exhibited acute tubular injury, podocyte injury and some crescent formation, and the main components of crescent are podocytes. Although renal tubules and vascular lesions gradually recover and crescent disappears, segmental lesions of podocyte appear in the late stage of RAS. These data reveal ultrastructural pathological changes during RAS, and suggest the role of podocyte lesions in chronic renal ischemia.


Diffraction-limited holography.
Experimental determinations of the size of the Airy disk for real image reconstructions of holograms of limited extent have yielded values comparable with those given by diffraction limitation. The holograms used are transmission, side-band holograms of low carrier frequency and are constructed in the far field of the object (~3-microm extent). In reconstruction, the real image is focused, with unit magnification, onto a rotating ground-glass plate and viewed through a microscope with 400x magnification. Comparisons of the reconstructed image with the actual object are made for holographic speeds ranging from f/10 to f/2 using visual, photographic, and photoelectric recordings. Photographic resolution tests, using standard bar patterns, yield a value of 1.05 for the ratio of the observed radius of the Airy disk to that corresponding to diffraction limitation. These tests are in disagreement with photoelectric measurements, the discrepancy being quantitatively ascribable to contrast enhancement in the film.


Logical intuition is not really about logic.
Recent research suggests that reasoners are able to draw simple logical or probabilistic inferences relatively intuitively and automatically, a capacity that has been termed "logical intuition" (see, e.g., De Neys & Pennycook, 2019). A key finding in support of this interpretation is that conclusion validity consistently interferes with judgments of conclusion believability, suggesting that information about logical validity is available quickly enough to interfere with belief judgments. In this study, we examined whether logical intuitions arise because reasoners are sensitive to the logical features of a problem or another structural feature that just happens to align with logical validity. In three experiments (N = 113, 137, and 254), we presented participants with logical (determinate) and pseudological (indeterminate) arguments and asked them to judge the validity or believability of the conclusion. Logical arguments had determinately valid or invalid conclusions, whereas pseudological arguments were all logically indeterminate, but some were pseudovalid (possible strong arguments) and others pseudoinvalid (possible weak arguments). Experiments 1 and 2 used simple modus ponens and affirming the consequent structures; Experiment 3 used more complex denying the antecedent and modus tollens structures. In all three experiments, we found that pseudovalidity interfered with belief judgments to the same extent as real validity. Altogether, these findings suggest that while people are able to draw inferences intuitively, and these inferences impact belief judgments, they are not logical intuitions. Rather, the intuitive inferences are driven by the processing of more superficial structural features that happen to align with logical validity. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Leaf-derived ABA regulates rice seed development via a transporter-mediated and temperature-sensitive mechanism.
Long-distance transport of the phytohormone abscisic acid (ABA) has been studied for ~50 years, yet its mechanistic basis and biological significance remain very poorly understood. Here, we show that leaf-derived ABA controls rice seed development in a temperature-dependent manner and is regulated by defective grain-filling 1 (DG1), a multidrug and toxic compound extrusion transporter that effluxes ABA at nodes and rachilla. Specifically, ABA is biosynthesized in both WT and dg1 leaves, but only WT caryopses accumulate leaf-derived ABA. Our demonstration that leaf-derived ABA activates starch synthesis genes explains the incompletely filled and floury seed phenotypes in dg1 Both the DG1-mediated long-distance ABA transport efficiency and grain-filling phenotypes are temperature sensitive. Moreover, we extended these mechanistic insights to other cereals by observing similar grain-filling defects in a maize DG1 ortholog mutant. Our study demonstrates that rice uses a leaf-to-caryopsis ABA transport-based mechanism to ensure normal seed development in response to variable temperatures.


Lyral is an important sensitizer in patients sensitive to fragrances.
Contact allergy to fragrances is a common problem world-wide. The currently used fragrance mix (FM) for patch testing has only eight constituents and does not identify all fragrance-allergic patients. As perfumes may contain 100 or more substances, the search for markers for allergy continues. The synthetic fragrance 4-(4-hydroxy-4-methylpentyl)-3-cyclohexene carboxaldehyde (Lyral) was tested together with the FM and 11 other fragrance substances on consecutive patients in six European departments of dermatology. All patients were carefully questioned regarding a history of reactions to scented products in the past and were grouped into four categories: 'certain', 'probable', 'questionable' and 'none'. Lyral (5% in petrolatum) gave a positive reaction in 2.7% of 1855 patients (range 1.2-17%) and ranked next to 11.3% with FM allergy. Twenty-four patients reacted to both Lyral and FM, but 21 (1.1%) reacted positively only to Lyral. Of 124 patients with a 'certain' history, 53.2% reacted to the FM and a further 7.2% to Lyral only. If any kind of history of fragrance intolerance was given, 80% (40 of 50) of Lyral positive patients had a 'positive' history while only 58.6% (123 of 210) of FM positive patients had such a history; this difference was significant at P < 0.01. Lyral was identified by gas chromatography-mass spectrometry in some products which had caused an allergic contact dermatitis in four typical patients who showed a patch test positive to Lyral and negative or doubtful to FM. In conclusion, we recommend the testing of 5% Lyral (in petrolatum) in patients suspected of contact dermatitis.


Treatment with apocynin selectively restores hippocampal arteriole function and seizure-induced hyperemia in a model of preeclampsia.
Preeclampsia (PE) is a hypertensive disorder of pregnancy associated with neurovascular dysfunction, cognitive impairment and increased seizure susceptibility. Here, we sought to determine if treatment of experimental PE (ePE) rats with apocynin could prevent hippocampal arteriolar (HA) dysfunction and impaired seizure-induced hyperemia within the hippocampus, a brain region central to cognition and seizure generation. Isolated and pressurized HAs from Sprague Dawley rats that were normal pregnant (Preg; n = 8), ePE (n = 8) or ePE treated with apocynin for 2 weeks of gestation (ePE + apo; n = 8) were compared. Hippocampal blood flow (n = 6/group) was measured using hydrogen clearance before and during seizure. Aorta elastin was quantified using histochemistry. ePE was associated with HA dysfunction including reduced contraction to endothelin-1 and diminished dilation to the endothelium-dependent vasodilator NS309 that was prevented by apocynin. However, apocynin had no effect on ePE-induced impairment of dilation to the nitric oxide donor sodium nitroprusside, but increased myogenic tone and substantially increased HA distensibility. Seizure-induced hyperemia was impaired in ePE rats that was restored by apocynin. Aorta from ePE rats had reduced elastin content, suggesting large artery stiffness, that was unaffected by apocynin. Thus, while apocynin partially prevented HA dysfunction, its restoration of functional hyperemia may be protective of seizure-induced injury during eclampsia.


Effect of praziquantel on pancreatic histopathological changes in experimental schistosomiasis mansoni.
The pancreatic tissue showed deposition of S. mansoni eggs on the 8th week of infection, multiple granulomata, interstitial oedema, inflammatory infiltrate, acinar atrophy and decrease in the size of islets of Langerhans. It was found that the most effective dose of praziquantel in such changes was 300 mg/kg/mouse, irrespective of being administered once or in three divided doses every four hours in the same day.


Comparison of Long-term Performance of Bioprosthetic Aortic Valves in Sweden From 2003 to 2018.
The performance of bioprosthetic aortic valves is usually assessed in single valve models or head-to-head comparisons. National databases or registries offer the opportunity to investigate all available valve models in the population and allows for a comparative assessment of their performance. To analyze the long-term rates of reintervention, all-cause mortality, and heart failure hospitalization associated with commonly used bioprosthetic aortic valves and to identify valve model groups with deviation in clinical performance. This population-based, nationwide cohort study included all adult patients who underwent surgical aortic valve replacement (with or without concomitant coronary artery bypass surgery or ascending aortic surgery) in Sweden between January 1, 2003, and December 31, 2018. Patients were identified from the SWEDEHEART (Swedish Web-System for Enhancement and Development of Evidence-Based Care in Heart Disease Evaluated According to Recommended Therapies) registry. Patients with concomitant valve surgery, previous cardiac surgery, and previous transcatheter valve replacement were excluded. Follow-up was complete for all participants. Data were analyzed from March 9, 2020, to October 12, 2021. Primary surgical aortic valve replacement with the Perimount, Mosaic/Hancock, Biocor/Epic, Mitroflow/Crown, Soprano, and Trifecta valve models. The primary outcome was cumulative incidence of reintervention, defined as a subsequent aortic valve operation or transcatheter valve replacement. Secondary outcomes were all-cause mortality and heart failure hospitalization. Regression standardization and flexible parametric survival models were used to account for intergroup differences. Mean follow-up time was 7.1 years, and maximum follow-up time was 16.0 years. A total of 16 983 patients (mean [SD] age, 72.6 [8.5] years; 10 685 men [62.9%]) were included in the analysis. The Perimount valve model group had the lowest and the Mitroflow/Crown valve model group had the highest cumulative incidence of reintervention. The estimated cumulative incidence of reintervention at 10 years was 3.6% (95% CI, 3.1%-4.2%) in the Perimount valve model group and 12.2% (95% CI, 9.8%-15.1%) in the Mitroflow/Crown valve model group. The estimated incidence of reintervention at 10 years was 9.3% (95% CI, 7.3%-11.3%) in the Soprano valve model group. Results of this study showed that the Perimount valve was the most commonly used and had the lowest incidence of reintervention, all-cause mortality, and heart failure hospitalization, whereas the Mitroflow/Crown valve had the highest rates. These findings highlight the need for clinical vigilance in patients who receive either a Soprano or Mitroflow/Crown aortic bioprosthesis.


Accuracy of Intraoperative Examination in Central Nervous System Lesions: A Study of 133 Cases.
Intraoperative examination is a highly valuable tool for the evaluation of central nervous system (CNS) lesions, helping the neurosurgeon to determine the best surgical management. This study aimed to evaluate the accuracy and to analyze the diagnostic disagreements and pitfalls of the intraoperative examinations through correlation with the final histopathological diagnosis in CNS lesions. Retrospective analysis of intraoperative examination of CNS lesions and their final diagnosis obtained during 16 consecutive years. All diagnoses were reviewed and classified according to World Health Organization (WHO) grading for CNS tumors. Squash was performed in 119 cases, while frozen section and both methods were done in 7 cases each. Among the 133 intraoperative examinations considered, 114 (85.7%) presented concordance and 19 (14.3%) diagnostic disagreement when compared with subsequent histopathological examinations. The sensitivity and specificity for the detection of neoplasia in intraoperative examination was 98 and 94%, respectively. The positive and negative predictive values were 99 and 88%, respectively. The accuracy for neoplastic and nonneoplastic disease was 85.7%. Disagreements were more frequent among low-grade (WHO grades I and II) neoplasms and nonmalignant cases. Our results showed good accuracy of the intraoperative assessments for diagnosis of CNS lesions, particularly in high-grade (grades III and IV) lesions and metastatic neoplasms.


DsrA regulatory RNA represses both hns and rbsD mRNAs through distinct mechanisms in Escherichia coli.
The 87 nucleotide long DsrA sRNA has been mostly studied for its translational activation of the transcriptional regulator RpoS. However, it also represses hns mRNA, which encodes H-NS, a major regulator that affects expression of nearly 5% of Escherichia coli genes. A speculative model previously suggested that DsrA would block hns mRNA translation by binding simultaneously to start and stop codon regions of hns mRNA (coaxial model). Here, we show that DsrA efficiently blocked translation of hns mRNA by base-pairing immediately downstream of the start codon. In addition, DsrA induced hns mRNA degradation by actively recruiting the RNA degradosome complex. Data presented here led to a model of DsrA action on hns mRNA, which supports a canonical mechanism of sRNA-induced mRNA degradation by binding to the translation initiation region. Furthermore, using MS2-affinity purification coupled with RNA sequencing technology (MAPS), we also demonstrated that DsrA targets rbsD mRNA, involved in ribose utilization. Surprisingly, DsrA base pairs far downstream of rbsD start codon and induces rapid degradation of the transcript. Thus, our study enables us to draw an extended DsrA targetome.


The distribution of neurokinin-1 and neurokinin-2 receptors in human central airways.
The precise locations of neurokinin (NK)-1 and NK-2 receptors in human airways, and their role in airway inflammatory diseases, have not been carefully examined. To determine the distribution of NK-1 and NK-2 receptors in human central airways, and to determine whether their distribution was different in smokers, we examined surgical specimens from patients undergoing lung resection for limited lung lesions. We mapped NK-1 and NK-2 receptors in four groups of subjects: four asymptomatic nonsmokers, seven asymptomatic smokers, seven symptomatic smokers with normal lung function, and eight symptomatic smokers with chronic airflow limitation. Tissues were immunostained with anti-NK-1- and anti-NK-2-receptor antibodies. Expression of NK-1 and NK-2 receptors was quantified through light microscopy and image analysis. Both NK-1 and NK-2 receptors were found in bronchial glands, bronchial vessels, and bronchial smooth muscle. Although no receptors were observed in the epithelium, receptors were occasionally found in nerves (NK-1) and in inflammatory cells (NK-2) such as T lymphocytes, macrophages, and mast cells. The distribution of both NK-1 and NK-2 receptors was similar in all the tissues examined in the four groups of subjects. These data show that NK-1 and NK-2 receptors are present in human central airways and that their expression is not modified by cigarette smoking.


Adverse remodeling in atrial fibrillation following isolated aortic valve replacement surgery.
Atrial fibrillation (AF) is the most common arrhythmia with adverse clinical outcomes. Aortic valve replacement (AVR) is one of the most frequently performed cardiac surgeries, although there is scarce evidence on arrhythmic outcomes. We aimed to evaluate AF during the first year post- isolated aortic valve replacement surgery and its clinical, analytical, and echocardiographic predictors. Severe aortic stenosis patients with no prior atrial fibrillation submitted to isolated aortic valve replacement surgery were included in our study, of which 316 remained in sinus rhythm and 24 developed AF. We performed logistic regression searching for AF predictors and a longitudinal comparison between pre and post-operative echocardiographic data. Postoperative AF (POAF), diabetes, and follow-up indexed Left Atrium Diameter (iLAD) were significantly higher in the group of patients developing AF. POAF and iLAD were independent AF predictors at follow-up. No differences between groups were found regarding baseline and follow-up echocardiographic data except for indexed Left Ventricle End-diastolic Diameter (LVED), which failed to decrease after surgery in the AF group. POAF and iLAD independently predicted AF at 1 year following isolated AVR surgery in aortic stenosis patients with no AF history. iLVED did not decrease significantly at follow-up in AF patients, possibly reflecting adverse ventricular remodeling.


Multiple roles of mucins in pancreatic cancer, a lethal and challenging malignancy.
Mucins are members of an expanding family of large multifunctional glycoproteins. Pancreatic mucins have important biological functions, including the protection, lubrication, and moisturisation of the surfaces of epithelial tissues lining ductal structures within the pancreas. Several lines of evidence support the notion that deregulated mucin production is a hallmark of inflammatory and neoplastic disorders of the pancreas. Herein, we discuss the factors that contribute to the lethality of pancreatic cancer as well as the key role played by mucins, particularly MUC1 and MUC4, in the development and progression of the disease. Aspects pertaining to the aberrant expression and glycosylation of mucins are discussed, with special emphasis on their potential impact on the design and implementation of adequate diagnostic and therapeutic strategies for combating this lethal malignancy.


HIV associated thrombotic microangiopathy.
Thrombotic microangiopathy (TMA) is a known complication of HIV infection. Endothelial cell injury appears to be the primary event causing platelet activation and deposition in the microvasculature. Direct cytopathic roles of HIV as well as other factors such as malignancy, drugs, and infectious agents have been implicated in the pathogenesis of HIV-TMA. Although the the majority of patients present in a more advanced stage of HIV disease, TMA can be the initial presenting symptom of HIV infection. Clinical features are those of idiopathic TMA, and the diagnosis should be suspected in any patient with new onset thrombocytopenia and microangiopathic haemolytic anaemia. Therapy with plasma exchange or infusion appears to be efficacious. A rapid diagnosis and institution of plasmapheresis is crucial for a favourable outcome. The long term prognosis of HIV-TMA is unfavourable and may depend on the stage of HIV infection. The recent data after the use of highly active retroviral treatment, however, are unavailable and current prognosis is therefore uncertain.


CARving the Path to Allogeneic CAR T Cell Therapy in Acute Myeloid Leukemia.
Despite advances in the understanding of the genetic landscape of acute myeloid leukemia (AML) and the addition of targeted biological and epigenetic therapies to the available armamentarium, achieving long-term disease-free survival remains an unmet need. Building on growing knowledge of the interactions between leukemic cells and their bone marrow microenvironment, strategies to battle AML by immunotherapy are under investigation. In the current review we describe the advances in immunotherapy for AML, with a focus on chimeric antigen receptor (CAR) T cell therapy. CARs constitute powerful immunologic modalities, with proven clinical success in B-Cell malignancies. We discuss the challenges and possible solutions for CAR T cell therapy development in AML, and examine the path currently being paved by preclinical and clinical efforts, from autologous to allogeneic products.


Elaboration and validation of Crohn's disease anoperineal lesions consensual definitions.
To establish consensual definitions of anoperineal lesions of Crohn's (APLOC) disease and assess interobserver agreement on their diagnosis between experts. A database of digitally recorded pictures of APLOC was examined by a coordinating group who selected two series of 20 pictures illustrating the various aspects of APLOC. A reading group comprised: eight experts from the Société Nationale Française de Colo Proctologie group of study and research in proctology and one academic dermatologist. All members of the coordinating and reading groups participated in dedicated meetings. The coordinating group initially conducted a literature review to analyse verbatim descriptions used to evaluate APLOC. The study included two phases: establishment of consensual definitions using a formal consensus method and later assessment of interobserver agreement on the diagnosis of APLOC using photos of APLOC, a standardised questionnaire and Fleiss's kappa test or descriptive statistics. Terms used in literature to evaluate visible APLOC did not include precise definitions or reference to definitions. Most of the expert reports on the first set of photos agreed with the main diagnosis but their verbatim reporting contained substantial variation. The definitions of ulceration (entity, depth, extension), anal skin tags (entity, inflammatory activity, ulcerated aspect), fistula (complexity, quality of drainage, inflammatory activity of external openings), perianal skin lesions (abscess, papules, edema, erythema) and anoperineal scars were validated. For fistulae, they decided to follow the American Gastroenterology Association's guidelines definitions. The diagnosis of ulceration (κ = 0.70), fistulae (κ = 0.75), inflammatory activity of external fistula openings (86.6% agreement), abscesses (84.6% agreement) and erythema (100% agreement) achieved a substantial degree of interobserver reproducibility. This study constructed consensual definitions of APLOC and their characteristics and showed that experts have a fair level of interobserver agreement when using most of the definitions.


COVID in Context: The Lived Experience of Richmond's Low-Income Older Adults.
Taking a phenomenological approach, this qualitative study describes the lived experiences of low-income older adults during the COVID-19 pandemic. A socio-ecological model was used to organize the five identified themes describing the lived experience: socio-economic context, Black Lives Matter and the politics of race, COVID and polarized views of COVID, interpersonal context (social connections), and individual context (feelings, beliefs, and behaviors). Study findings illustrate the intersectionality of contextual influences on the experience of low-income older adults. Study participants demonstrated remarkable resilience and coping strategies developed in response to the challenges they experienced throughout their lifetime which benefited them when faced with the pandemic, social unrest, and political events that took place in 2020. This study highlights the importance of understanding the larger context of COVID-19 which has significant implications for policy makers and public health leaders.


Evidence for acute tolerance to the behavioral effects of lindane: concomitant changes in regional monoamine status.
The effects of the organochlorine insecticide lindane on plus-maze ((+)-maze) behavior of rats and on regional monoamine status were studied at two time points, 30 min and 24 hr post-dosing. Animals were given lindane, 20 mg/kg, 30 min before (L1/2 group), or 40 mg/kg, 24 hr before (L24 group), experimental time; these schedules allowed the study of animals with equivalent brain concentration of lindane at two different time points after administration. The (+)-maze results indicated a reduction in the number of entries into the arms of the maze 30 min after administration of lindane that was not present 24 hr later, suggesting the development of an acute tolerance to the behavioral effects of the chemical. In a parallel group of animals, concentrations of noradrenaline (NA), serotonin (5HT), 5-hydroxyindoleacetic acid (5HIAA), dopamine (DA), dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) were determined by HPLC in seven brain regions. Significant decreases in NA concentrations were found in L1/2 animals in hippocampus and cerebral cortex and also in the L24 group in the latter region. 5HT/5HIAA ratios increased in several brain regions in the L24 rats but not in the L1/2 rats, thus showing an inverse relationship with behavioral effects. DOPAC/DA and HVA/DA ratios in hypothalamus and cerebral cortex were, by contrast, increased in both groups of treated animals. In conclusion, it appears that an acute tolerance can develop to the behavioral effects of lindane, and that there are modifications produced in central monoaminergic systems by lindane that show brain region and and treatment schedule specificity.(ABSTRACT TRUNCATED AT 250 WORDS)


Microsurgical anatomy of the anterior cerebral artery in Indian cadavers.
The microanatomy features of cerebral arteries may be variable and may be different in different ethnic groups. To study the anterior cerebral artery (ACA) anatomy in North-West Indian cadavers. Microanatomy features of the ACA were studied in 15 formalin fixed human cadaveric brains under microscope. The outer diameter, length, and number of perforating branches with respective anomalies were measured for each of the following vessels: ACA (proximal A1 segment to distal A2 segment), anterior communicating artery (ACoA), Recurrent artery of Heubner (RAH), and callosomarginal artery and photographed for documentation. The mean length and external diameter of right and left A1 segment was 12.09 mm and 12.0 mm and 2.32 mm and 2.36 mm respectively. Narrowing, duplication, and median ACA were seen in 6.6%, 3.3% and 6.6% of the vessels respectively. Complex ACoA type was seen in 40% cadavers. RAH originated at an average point of 0.2 mm distal to ACoA, but in one cadaver it arose 5 mm proximal to ACoA. Double RAH was found in 26.6%. The course of RAH in relation to A1 was superiorly in 60%, in anteriorly 30% and posteriorly in 10% of cadavers. The orbitofrontal artery (OFA) and frontopolar artery (FPA) arose from A2 in 83.3% to 40% respectively. The mean distance of OFA and FPA from ACoA was 4.17 mm and 8.5 mm respectively. After giving rise to central, callosal and cortical branches, pericallosal artery terminated near the splenium of the corpus callosum or on the precuneus as the inferomedial parietal artery. Knowledge of the microvascular anatomy is indispensable and it is mandatory to be aware of the possible variations in the anomalies to minimize morbidity.


Children's coordination of force output in a pinch grip task.
This study examined the role of sensorimotor system noise in the organization of the force output of the thumb and index finger and the coordination between the two digits in an isometric pinch grip force task as a function of age (6, 8, 10, 18-22 years), feedback condition (with and without visual feedback information), and force level (5, 15, 25, and 35% of maximal voluntary force. With increases in age under the visual feedback conditions, the signal-to-noise ratio increased, the sequential structure of the force output signals became more irregular, the degree of coherence between the digits at higher force levels was enhanced, and the digits exhibited a greater degree of coherence across the higher frequencies of the power spectrum at all force levels. However, these age differences were either minimized or eliminated under conditions without feedback. These findings show that the age-related performance differences in grip force variability are primarily due to more effective use of visual information rather than age differences in intrinsic sensorimotor noise.


Dental Health Status of HIV-Positive Patients and Related Variables in Southeast Iran.
Different factors can be responsible for the increased prevalence of dental caries and missing teeth in HIV-positive patients. This study evaluates dental health status and its relationship with social, behavioral, and medical factors in HIV-positive patients under the coverage of Zahedan University of Medical Sciences in Southeast Iran. In a cross-sectional study, the dental health status of 119 HIV-positive patients was assessed in accordance with WHO indices and included decayed, missing, and filled teeth (DMFT). A questionnaire on different social, behavioral, and medical variables was filled out for every case and the relationship and correlation of the variables to dental health status were investigated using One-way ANOVA, the Kruskal Wallis test, the t-test, the Mann-Whitney test, Spearman's rho correlation coefficient, and Pearson correlation. The mean value of DMFT index was 11.87 ± 8.08, where the mean values of decayed and missing teeth were 8.42 ± 5.44 and 3.43 ± 4.07, respectively. DMFT index, decayed, and missing teeth correlated only with age (P < 0.0001, P = 0.009, P < 0.0001) and duration of HIV involvement (P = 0.004, P = 0.031, P = 0.007). The dental health status of HIV-positive patients in this region was almost inappropriate. Most social, behavioral, and medical factors had no influence on dental health; only a correlation between dental health, age, and duration of HIV involvement was observed.


In vivo quantification of ageing changes in the rat liver from early juvenile to senescent life.
Using high resolution multifluorescence in vivo microscopy, the present study was undertaken to determine the changes in rat hepatic tissue architecture and microvasculature during the growth associated with juvenile maturation and adult senescence, i.e. the age of 1, 3, 12 and 24 months. By 1 month of age the liver attained its full size and functional capacity, as assessed by relative organ weight and hepatic bile flow. Survey of liver architecture revealed a progressive growth of lobular area with postsinusoidal venules exhibiting a proportional increase in length, diameter and inter-vascular distance up to the age of 12-24 months. In regard to the 3.5-4-fold average increase of lobular units, a minor reduction of sinusoidal density to 87% over life strongly implies the recruitment or formation of new sinusoidal microvessels as contributing mechanism to meet oxygen demand due to overall tissue enlargement. The sinusoidal perfusion rate remained above 98% over the whole lifespan. Leukocytic interaction with the hepatic microvascular endothelium was found within the physiological range in all age groups. Moreover, kinetics of clearance of latex beads as well as lobular distribution of Kupffer cells did not differ between animals of different age. Hepatic stellate cell-associated area of ultraviolet vitamin A-autofluorescence increased with age and significantly correlated with increasing tissue concentrations of vitamin A metabolites. Biochemical parameters serving as measures of tissue integrity did not indicate age-associated tissue alterations. These age-associated physiological changes should be carefully taken into account as a relevant variable in experimental research.


Responding to change: how did bar workers adapt to the smoke-free legislation in Scotland?
In recent years a number of countries have implemented comprehensive smoking bans to protect workers from adverse effects of exposure to second-hand smoke. It is also well known that many key stakeholders working in hospitality settings are themselves smokers. This paper reports the findings of a longitudinal qualitative study conducted with bar workers following the implementation of smoke-free legislation in Scotland. It examines the effects of smoking prevalence and status on support for smoke-free legislation, and its impact on business and working practices as well as bar workers' smoking behaviour both in and outside of the workplace. The results highlight a need for targeted assistance for businesses serving disadvantaged communities both to adapt commercially to the legislation and to provide cessation support to workers who smoke.


Usefulness of the hsp60 gene for the identification and classification of Gram-negative anaerobic rods.
The hsp60 gene sequences were determined for 121 strains of Gram-negative anaerobic rods, including the genera Bacteroides, Barnesiella, Butyricimonas, Odoribacter, Parabacteroides, Paraprevotella, Porphyromonas, Prevotella and Tannerella. The mean pairwise hsp60 gene sequence similarity (73.8-97.1 %) between species in each genus, except for the genus Tannerella that comprises one species, was significantly less than that of the 16S rRNA gene sequence (88.3-96.3 %). Only pairwise hsp60 gene sequence similarity (97.1 %) of the genus Paraprevotella was higher than that of the 16S rRNA gene sequence (93.8 %). Each genus formed a distinct clade in the phylogenetic analysis of the hsp60 gene sequence as well as the 16S rRNA gene sequence. The phylogenetic analysis indicated a higher evolutionary rate for the hsp60 gene sequence than the 16S rRNA gene sequence, especially in the genera Porphyromonas and Prevotella. This study suggests that the hsp60 gene is a useful alternative phylogenetic marker for the identification and classification of a broad range of Gram-negative anaerobic rods.


Potentiation of excito-toxicity by glutamate uptake inhibitor rather than glutamine synthetase inhibitor.
The neuroprotective functions of glia cells in the presence of excessive amounts of extracellular glutamate (Glu) were examined using glia-rich and glia-poor cultured cerebellar granule cells that contained the same number of neurons. In order to focus on the metabolic enzyme glutamine synthetase (GS) and the uptake system in glia cells, selective inhibitors such as L-methionine sulfoximine (MSO) and 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) were used as pharmacological tools. The increased amount of lactate dehydrogenase (LDH) leakage induced by 50 microM Glu and SITS was equivalent to that of 1 mM Glu. However, the simultaneous treatment with 50 microM Glu and 5 microM MSO did not increase the LDH leakage. The larger quantities of extracellular Glu were sustained in both glia-rich and glia-poor cultures. After the administration of Glu and MSO, however, the larger quantities of Glu were not sustained. Taking these results into consideration, the Glu uptake system in glia cells seems to be more important than the Glu metabolic enzyme system in the regulation of neuronal protection from Glu toxicity.


Effect of race on colon cancer treatment and outcomes in the Department of Defense healthcare system.
The increase in mortality noted in African Americans with colon cancer is attributed to advanced stage at presentation and disparities in treatment received. The aim of this study was to assess the influence of race on the treatments and survival of colon cancer patients in an equal-access healthcare system. This retrospective cohort study included African American and white patients with colon cancer treated at Department of Defense facilities. Disease stage, surgery performed, chemotherapy used, and overall survival were evaluated. Of the 6958 colon cancer patients identified, 1115 were African American. African Americans presented more frequently with stage IV disease, 23% vs 17% for whites (P < .001). There was no difference in surgical resection rates for African American or whites (85.8% vs 85.5%, respectively; chi2, P > .05). There was no difference in the use of systemic chemotherapy for stage III colon cancer (73.5% for African Americans vs 72.2% for whites; chi2, P > .05) or stage IV colon cancer (56.3% for African Americans vs 54.4% for whites; chi2, P > .05). The overall 5-year survival rate was similar for African American and white patients (56.1% vs 58.5%, respectively; log-rank, P > .05). After adjusting for gender, age, tumor grade, and stage, African American race was not a risk factor for survival in Cox proportional hazard analysis (hazard ratio, 0.981; 95% confidence interval, 0.888-1.084). In an equal-access healthcare system, African American race is not associated with an increase in mortality. African American patients undergo surgery and chemotherapy is administered at rates equal to whites for all stages of colon cancer.


[Thermosensitivity of glioma cells with special reference to changes in cytoskeletons].
The relationship between the thermosensitivity of cultured brain tumor cells and cytoskeleton was studied. C6 rat glioma cell line (C6 cells) and U-373-MG human glioblastoma cell line (MG cells) were used in monolayer culture. Survival rates at various temperatures were calculated by colony forming assay 10 days after heat treatment. Actin filaments, the main components of microfilaments, were observed by the 7-chloro-4-nitrobenzo-2-oxadiazole phallacidin staining and indirect immunofluorescence staining methods. Alpha-tubulins, the main components of microtubules, were also stained with an indirect immunofluorescence staining method. The morphological changes were investigated by scanning electron microscopy (SEM). Both the C6 cells and the MG cells showed moderate thermosensitivity on the survival curves. Actin filaments were revealed at stress fibers and the ruffles of the leading edge on both cell lines. Stress fibers were well developed in MG cells but were only minor in C6 cells. After heat treatment ruffles and stress fibers were disrupted. However, alpha-tubulins were not affected by heat treatment. SEM showed Swiss-cheese like change of cell surfaces due to many pores with disruption of ruffles and stress fibers after heat treatment. These results suggest that the cytoskeleton, especially microfilaments, may be damaged by hyperthermia.


Molecular cloning and nucleotide sequence of the nuclear PET122 gene required for expression of the mitochondrial COX3 gene in S. cerevisiae.
The nuclear PET122 gene from S. cerevisiae is necessary for translation of a single mitochondrial mRNA that encodes subunit III of cytochrome c oxidase. We report here the cloning and nucleotide sequence of PET122, and properties of the predicted protein product, which consists of 242 residues. Analysis of PET122-lacZ translational fusions confirms that the PET122 coding region is translated in vivo and indicates that the PET122 protein product is targeted to mitochondria. A 117 residue domain located in the carboxy-terminal half of the PET122 protein, at least part of which is shown by characterization of mutants to be critical for PET122 function, exhibits 24% identity and 59% similarity to a portion of the catalytic domain of E. coli alanyl-tRNA synthetase. However, pet122 mutants are not defective in mitochondrial translation per se, as would be expected if PET122 encoded a tRNA synthetase. Instead, the PET122 protein may carry out one or more activities in common with tRNA synthetases, such as binding of ATP or RNA.


Heavy vehicle driver fatalities: learning's from fatal road crash investigations in Victoria.
This study describes the nature and extent of fatal heavy vehicle driver crashes in Victoria between 1999 and 2007 and the factors associated with the crash. A descriptive study was conducted comprising the population of heavy vehicle drivers killed in a road transport crash while operating a vehicle of > or =4.5 tonne Gross Vehicle Mass (GVM) for the purposes of work. Information about the nature of crash, environmental, driver, vehicle and occupational factors were collected from the coroner's death investigation file. Of the 91 deaths identified 61 were eligible for review. All 61 cases were male, solo drivers with a mean age of 44.7 years. Most vehicles were articulated in configuration. One-third of crash scenarios involved a single vehicle leaving the roadway on a straight road. One in every six fatally injured drivers was detected with the presence of stimulants or cannabis. Twenty-two drivers were travelling at excessive or inappropriate speeds for the circumstances. Seatbelt wearing status was documented for only 25 of 61 drivers. This study is the first to comprehensively examine a population of fatally injured heavy vehicle drivers using coroner's investigation files and these findings are consistent with previous, less detailed, Australian research. Information about potential associations between occupational factors and crash risk was limited. Improving driver safety requires incorporation of occupational factors into a standardised approach to heavy vehicle crash investigations.


Effects of Wine and Tyrosol on the Lipid Metabolic Profile of Subjects at Risk of Cardiovascular Disease: Potential Cardioprotective Role of Ceramides.
Ceramides are a class of sphingolipids which have recently been shown to be better cardiovascular disease (CVD) risk predictors than traditional CVD risk biomarkers. Tyrosol (TYR) is a dietary phenolic compound known to possess cardioprotective effects per se or through its in vivo active metabolite hydroxytyrosol. The purpose of this study was to evaluate the effects of the co-administration of white wine (WW) and TYR on circulating levels of ceramides and other lipids in humans at high CVD risk. Volunteers underwent a randomized controlled crossover clinical trial (4-week duration per intervention) with three different interventions: control, WW, and WW enriched with a capsule of TYR (WW + TYR). Endothelial function cardiovascular biomarkers and plasma lipidomic profile were assessed before and after each intervention. It was found that the WW + TYR intervention resulted in lower levels of three ceramide ratios, associated with an improvement of endothelial function (Cer C16:0/Cer C24:0, Cer C18:0/Cer C24:0, and Cer C24:1/Cer C24:0), when compared to the control intervention. Moreover, WW + TYR was able to minimize the alterations in plasma diacylglycerols concentrations observed following WW. Overall, the results obtained show that the antioxidant TYR administered with WW exerts beneficial effects at the cardiovascular level, in part by modulating blood lipid profile.


Anomalous observations of hydrocodone in patients on oxycodone.
Urine drug monitoring is used by physicians treating chronic pain patients with opioid therapy. Patients are tested in part to insure that they are not taking other drugs. Therefore, the finding of hydrocodone in a patient who is only prescribed oxycodone has clinical implications. Oxycodone preparations are known to have small amounts of hydrocodone as an impurity estimated to be < 0.1%. We established the concentration of unexpected hydrocodone in patients taking oxycodone. Urine drug testing specimens from a population of 30,000 pain patients prescribed oxycodone in various formulations were quantitatively measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The frequency and concentration of hydrocodone as a function of oxycodone concentration were determined. There were 187 specimens with > 100,000 ng/ml of oxycodone. Of these, 72% were positive for hydrocodone. Of the 311 specimens with oxycodone concentrations > 50,000-100,000 ng/ml, 33% were positive for hydrocodone. Of the 1067 specimens with oxycodone > 20,000-50,000 ng/ml, 16% were positive for hydrocodone. Of the 8508 specimens with oxycodone > 1000-20,000 ng/ml, 16% were positive for hydrocodone. The high frequency of hydrocodone in samples containing high concentrations of oxycodone was ascribed to the manufacturing process of the oxycodone medications. However, a significant number of patients also took hydrocodone that was not listed on their prescribed medications. When oxycodone is > 100,000 ng/ml, hydrocodone should be <1500 ng/ml. When oxycodone is < 100,000 ng/ml then hydrocodone should be <500 ng/ml. Values greater than these indicate non-prescribed hydrocodone use. Clinicians and laboratories testing urine for drugs should be aware of the possibility of low concentrations of hydrocodone in the urine of patients taking high doses of oxycodone.


Repair of DNA damage induced by bile salts in Salmonella enterica.
Exposure of Salmonella enterica to sodium cholate, sodium deoxycholate, sodium chenodeoxycholate, sodium glycocholate, sodium taurocholate, or sodium glycochenodeoxycholate induces the SOS response, indicating that the DNA-damaging activity of bile resides in bile salts. Bile increases the frequency of GC --> AT transitions and induces the expression of genes belonging to the OxyR and SoxRS regulons, suggesting that bile salts may cause oxidative DNA damage. S. enterica mutants lacking both exonuclease III (XthA) and endonuclease IV (Nfo) are bile sensitive, indicating that S. enterica requires base excision repair (BER) to overcome DNA damage caused by bile salts. Bile resistance also requires DinB polymerase, suggesting the need of SOS-associated translesion DNA synthesis. Certain recombination functions are also required for bile resistance, and a key factor is the RecBCD enzyme. The extreme bile sensitivity of RecB-, RecC-, and RecA- RecD- mutants provides evidence that bile-induced damage may impair DNA replication.


High-resolution solution structure of the EGF-like domain of heregulin-alpha.
The solution structure of the 63-residue heregulin-alpha (HRG-alpha) epidermal growth factor (EGF)-like domain, corresponding to residues 177-239 of HRG-alpha, has been determined to high resolution using data from two-dimensional and three-dimensional homo- and heteronuclear NMR spectroscopy. The structure is based on a total of 887 internuclear distance and dihedral restraints derived from data obtained using unlabeled and uniformly 15N-labeled protein samples, at pH 4.5, 20 degrees C. A total of 20 structures were calculated using a hybrid distance geometry-simulated annealing approach with the program DGII, followed by restrained molecular dynamics using the program DISCOVER. The average maximum violations are 0.12 +/- 0.01 angstroms and 1.4 +/- 0.3 degrees for distance and dihedral restraints, respectively. The backbone (N,C(alpha),C) atomic rms distribution about the mean coordinates for residues 3-23 and 31-49 is 0.29 +/- 0/07 angstroms. The N-and C-terminal residues (1-2 and 50-63) and 24-30 are disordered. Comparison of the HRG-alpha EGF-like domain structure with the previously determined structure of human EGF [Hommel et al. (1992) J. Mol. Biol. 227, 271-282] reveals a high degree of structural similarity; excluding the N-terminal region (residues 1-13), the disordered phi-loop region (residues 24-30) that contains a three-residue insertion in HRG-alpha relative to hEGF, and the disordered C-terminal region (residues 50-63), the C(alpha) alignment between the HRG-alpha and hEGF minimized mean structures has a rms difference of approximately 1 angstrom. In HRG-alpha the N-terminal residues 2-6 form a well-defined beta strand rather than being disordered as found for hEGF. This structural difference correlates with functional data which suggest that the N-terminal region of the HRG-alpha EGF-like domain is responsible for the observed receptor specificity differences between HRG-alpha and EGF.


Homoserine esterification in green plants.
EXTRACTS OF PHYLOGENETICALLY DIVERSE PLANS WERE SURVEYED FOR THEIR ABILITY TO SYNTHESIZE THE FOLLOWING HOMOSERINE ESTERS WHICH ARE POTENTIAL PRECURSORS FOR METHIONINE AND THREONINE SYNTHESIS IN GREEN PLANTS: O-acetyl-, O-oxalyl-, O-succinyl-, O-malonyl-, and O-phosphohomoserine. Synthesis of O-acylhomoserine esters was detected only in Pisum sativum L. and Lathyrus sativus L. Extracts of P. sativum, a plant known to accumulate O-acetylhomoserine, catalyzed the specific synthesis of this ester from homoserine and acetyl-CoA. Extracts of L. sativus, a plant known to accumulate O-oxalylhomoserine, catalyzed the specific synthesis of this ester from homoserine and oxalyl-CoA. None of the other plants surveyed, including representatives of the green algae, horsetails, gymnosperms, and angiosperms, catalyzed the synthesis of any of the O-acylhomoserine esters studied. In contrast, synthesis of O-phosphohomoserine by the reaction catalyzed by homoserine kinase was demonstrated in extracts of all plants examined, including the two exceptional legumes.These results suggest that, among the five homoserine esters studied, O-phosphohomoserine is the major activated homoserine derivative in plants. Direct confirmation of the dominant physiological role of O-phosphohomoserine in the synthesis of cystathionine in the transsulfuration pathway of methionine biosynthesis in plants has recently been provided (Datko, A. H., Giovanelli, J., and Mudd, S. H. 1974. J. Biol. Chem. 249: 1139-1155).


Nitrate Assimilation and Crassulacean Acid Metabolism in Leaves of Kalanchoë fedtschenkoi Variety Marginata.
The enzymes necessary to assimilate ammonia either via glutamine synthetase and glutamate synthase or via the glutamate dehydrogenase pathways are present in both green and white leaf tissues of Kalanchoë fedtschenkoi. Nitrate reductase activity develops to a maximum in a Crassulacean acid metabolism (CAM) plant canopy before either ribulose 1,5-bisphosphate carboxylase, or phosphoenolpyruvate carboxylase, or CAM. Nitrate reductase also is activated each morning and is inactivated late in the day as in other plants. However, there does not appear to be any direct relationship between nitrate reductase activity and the level of acid, its daily pattern or the amplitude of CAM. Though nitrate reductase is activated maximally each day by light, in Kalanchoë leaves for six days the activity followed a precise daily pattern independent of continuous light or dark.


Nicotine patch vs. nicotine lozenge for smoking cessation: an effectiveness trial coordinated by the Community Clinical Oncology Program.
Nicotine replacement therapies are efficacious for treating nicotine dependence. However, limited data exist on benefits of different NRTs and predictors of treatment outcome. This study compared the effectiveness of transdermal nicotine vs. nicotine lozenge for smoking cessation and identified predictors of treatment response. A randomized, open-label effectiveness trial was conducted at 12 medical sites participating in the National Cancer Institute's Community Clinical Oncology Program. The sample consisted of 642 treatment-seeking smokers randomized to 12 weeks of transdermal nicotine or nicotine lozenge. Smoker characteristics were assessed at baseline, and 24-h point prevalence abstinence confirmed with breath carbon monoxide (CO) was evaluated at end of treatment (EOT) and at a 6-month follow-up. There was a trend for higher quit rates for transdermal nicotine vs. nicotine lozenge at EOT (24.3% vs. 18.7%, p=.10) and 6 months (15.6% vs. 10.9%, p=.10). A logistic regression model of EOT quit rates showed smokers who preferred transdermal nicotine, were not reactive to smoking cues, and did not use nicotine to alleviate distress or stimulate cognitive function had higher quit rates on transdermal nicotine. A logistic regression model of 6-month quit rates showed smokers who preferred transdermal nicotine had higher quit rates on transdermal nicotine, and smokers who used nicotine to alleviate distress or stimulate cognitive processes had lower quit rates on nicotine lozenge. Transdermal nicotine may be more effective than nicotine lozenge for smokers who prefer transdermal nicotine and do not smoke to alleviate emotional distress or stimulate cognitive function.


[Proportion of proinsulin two minutes after arginine stimulation: a pilot study].
To seek an easy way to evaluate islet beta cell function. 214 residents of Shanghai, aged 20-70, all of Han nationality, were divided into 3 groups according the 1999 WHO standard: 39 in the normal glucose tolerance (NGT) group, aged 52 years +/- 6 years, with the BMI of 22.3 kg/m(2) +/- 0.4 kg/m(2); 29 in the impaired glucose tolerance (IGT) group, aged 54 years +/- 9 years, with the BMI of 24.4 kg/m(2) +/- 0.6 kg/m(2); and 146 in the T2DM group, that was subdivided into 2 subgroups: 43 in the newly diagnosed T2DM (NT2DM) group, aged 52 years +/- 9 years, with the BMI of 24.4 kg/m(2) +/- 0.5 kg/m(2), and 103 in the previously diagnosed T2DM (PT2DM) group, aged 57 years +/- 9 years, with the BMI of 23.5 kg/m(2) +/- 0.3 kg/m(2). Arginine at the dose of 10% 50 ml was injected intravenously in fasting situation. Two, four, and six minutes after blood samples were collected from a needle indwelling in the contralateral vein to examine the levels of plasma glucose (PG), true insulin (TI), and proinsulin (PI). (1) The TI and TI/PG levels of all groups peaked 2 minutes after arginine stimulation and then gradually decreased, however, the levels 6 minutes after stimulation were still higher than the baseline values. The peak values of TI and TI/PG were arranged from high to low in the order of IGT, NGT, NT2DM and PT2DM. The peak values of NT2DM and PT2DM groups were significantly lower than those of the NGT and IGT groups (all P < 0.01). The peak values of the NT2DM group were significantly higher than those of the PT2DM group (both P < 0.01). (2) The PI value of the IGT, NT2DM and PT2DM groups peaked 2 minutes after the stimulation and then gradually decreased with the peak values significantly higher than that of the NGT group (all P < 0.05). The fasting PI/TI and PI/(TI + PI) of the NGT group were all significantly lower than those of the IGT group (both P < 0.05). (3) The changes of PI/TI in each groups manifested almost the mirror image of TI/PG mentioned above, and the lowest PI/TI levels were observed 2 minutes after stimulation. The similar changes of PI/(TI + PI) were found. The lowest points of groups 2 minutes after stimulation was arranged from low to high in the order of NGT, IGT, NT2DM and PT2DM. The PI/TI and PI/(TI + PI) values 2 minutes after arginine stimulation may be used as predictors for beta cell function in clinical practice.


Cyclic Nucleotide (cAMP and cGMP) Assays and Capture ELISA for Quantitative Analysis of Plasmodium falciparum Blood-stage Egress.
Upon rupture of Plasmodium falciparum (P. falciparum) schizonts in vitro (an event known as egress), merozoites are released into the culture medium. The merozoites invade fresh red blood cells, a process that involves shedding of a microneme protein called apical membrane antigen-1 (AMA1) from the merozoite surface. This shedding, which takes place even in the absence of invasion, is therefore a surrogate marker for the degree of egress taking place in a culture, and can be measured using a specific capture ELISA to quantify AMA1 levels in culture supernatants (Collins et al., 2013). The assay uses a monoclonal antibody specific for AMA1 (called 4G2dc1) (Kocken et al., 1998; Collins et al., 2009) to capture and immobilize the protein from culture supernatants, then uses a specific rabbit polyclonal antiserum to detect the immobilized antigen. A phosphatase-conjugated goat anti-rabbit antibody is finally used to quantify the binding of the second antibody. Egress is absolutely dependent upon the activity of a parasite cGMP-dependent protein kinase, PKG, and so is influenced by levels of intracellular cGMP (Collins et al., 2013). This is regulated by the interplay between guanylate cyclases and phosphodiesterases. The latter enzymes may also degrade cAMP, so it may also be informative to measure intracellular cAMP levels.


Lipopolysaccharide pretreatment of cyclosporine-treated rats enhances cardiac allograft survival.
Lipopolysaccharide (LPS), the endotoxin in gram-negative bacterial cell walls, is a major factor in septic shock. Tumor necrosis factor (TNF) appears immediately after LPS release or LPS injection in rats, but when these animals have LPS reinjected for up to 7 days, TNF production is inhibited. Because inhibiting TNF with anti-TNF antibodies prolongs cardiac allograft survival and is synergistic with cyclosporine (CsA), enhanced graft survival could result from inhibiting TNF via LPS pretreatment. Accordingly, heterotopic rat heart transplants were performed in: I, untreated controls: II, LPS pretransplant treatment: III, LPS post-transplant treatment; IV, low-dose CsA post-transplant treatment; V, CsA post-transplant treatment and PBS (LPS vehicle); or VI, LPS pretransplant treatment and low-dose CsA post-transplant treatment, using Brown Norway (BN) donors and Lewis (LEW) recipients. Rejection was defined by a lack of contractions. Results showed that while LPS pre- or post-treatment alone had little allograft survival effect, LPS pretreatment combined with CsA significantly prolonged survival vs control or CsA alone (22.0 +/- 1.6 days vs 6.8 +/- 0.6 days or 13.4 +/- 1.1 days; P < 0.001). Primary MLRs of LPS-pretreated LEW splenocytes cocultured with irradiated BN splenocytes had significantly less [3H]thymidine incorporation than untreated LEW splenocytes (3671 +/- 349 vs 7828 +/- 14 cpm). TNF assays of untreated and PBS-treated LEW spleen cells cocultured with irradiated BN spleen cells had 1.3 and 1.1 pg of TNF/10(6) cells, respectively, in 2 hr, but no TNF from LPS-pretreated LEW cells was detected.(ABSTRACT TRUNCATED AT 250 WORDS)


[Lectins in pulmonary adenocarcinomas].
A relatively specific binding of lectins to various glycids enabled authors to evaluate the exosecretion characteristic of lung carcinomas closer than by HE, investigation of mucins and immunohistochemical epithelial markers. The main subtypes of lung adenocarcinomas (usual of acinar, tubulopillary cubocellular or cylindrocellular, solid with mucus production), pseudosarcoma, large cell and undifferentiated carcinomas from 9 bioptical samples were therefore compared as to binding of 5 lectins with preferential affinity to glucose/mannose (CON A), acetylglucosamin (WGA) and acetylgalactosamin (PNA, RCA, HPA). All the subtypes of adenocarcinoma as well as undifferentiated carcinoma showed a strong binding of bean lectin (CON A) and a slighter binding of ricinus (RCA) and wheat germ (WGA) lectins. Binding of Helix pomatia lectin (HPA) was nearly parallel to that of CON A even in large cell carcinoma (without mucin positivity)-except in undifferentiated carcinoma (in HE reminding of squamous carcinoma but CK negative). Peanut lectin (PNA) binding correlated with the production of acid glycosaminglycans and its lacking might serve as an indirect sign of Clara cell origin in some cubocellular bronchioloalveolar carcinomas which was otherwise difficult to prove. Lections mostly did not bind to mucus vacuoles and cytoplasmic granulary positivities represented secretion granules; marginal membranous positivities represented glycocalyx or lipoproteinaceous layer released by Clara cells in accordance with the expression of EMA. In pseudosarcoma a gradual binding of CON A from sarcomatoid to epithelial areas allowed to evaluate their connection better than according to an expression of cytokreatin.


The influence of exocyclic substituents of purine bases on DNA curvature.
Complementary oligonucleotides with 5' overhanging deoxyguanosine or deoxycytidine stretches, respectively, of the general form 5'-d(GGGCAARAAC).5'-d(CCCGTTYTTG), where R represents the bases adenine (A), hypoxanthine (base of inosine nucleoside, I), purine (R), 2-aminopurine (n2R), or 2,6-diaminopurine (n2,6(2)R) and where Y represents the pyrimidine bases thymine (T) or cytosine (C), have been chemically synthesized. After hybridization of complementary fragments, they were ligated to form multimers and analyzed by polyacrylamide gel electrophoresis. Anomalous gel migration was observed for the sequences 5'-d(AARAA) when the R.Y base pair was dA.dT, dI.dC, or dR.dT. All of these base pairs lack at least the amino group at position 2 of the purine base. The degree of anomalous gel migration was also related to the substituent at position 6 of the purine base. An amino group at position 6 was more effective than a carbonyl or a hydrogen in inducing anomalous gel migration. Additionally, the fragments 5'-d(GGGCAIAIAC).5'-d(CCCGTCTCTG), 5'-d(GGGCAIIIAC).5'-d(CCCGTCCCTG), and 5'-d(GGGCIIAIIC).5'-d(CCCGCCTCCG) were prepared in which increasing numbers of dA.dT base pairs are replaced by dI.dC base pairs. The degree of gel-migration anomaly of these sequences correlates with the number of dA.dT base pairs left in the five-base purine block. The data support the hypothesis that within the deoxyadenosine tracts, the base pairs fold into the minor groove at position 2 of the base to balance for the NH2 groups at position 6. This hypothesis explains the formation of a B'-form DNA structure for the deoxyadenosine tracts as well as DNA curvature.


Ibudilast inhibits cerebral aneurysms by down-regulating inflammation-related molecules in the vascular wall of rats.
Phosphodiesterase-4 (PDE-4) is a cyclic adenosine monophosphate-specific enzyme involved in various inflammatory diseases. We studied its role in and the effect of ibudilast, which predominantly blocks PDE-4, on rat cerebral aneurysms. Cerebral aneurysms were induced at the anterior cerebral artery-olfactory artery bifurcation of female rats subjected to hypertension, increased hemodynamic stress, and estrogen deficiency. The effect of ibudilast (30 or 60 mg/kg/d for 3 months) on their cerebral aneurysms was studied by morphological and immunohistochemical assessment and quantitative real-time polymerase chain reaction assay. In our in vitro study, we grew endothelial cells stimulated by angiotensin II under estrogen-free conditions and examined the effect of ibudilast on PDE-4 activation and the cyclic adenosine monophosphate level. Morphological evaluation using vascular corrosion casts showed ibudilast significantly suppressed cerebral aneurysms in a dose-dependent manner. In rats with induced cerebral aneurysms, the gene and protein expression of PDE-4 was high, and endothelial leukocyte adhesion molecules (P-selectin, intracellular adhesion molecule 1, and vascular adhesion molecule 1), matrix metalloproteinase-9, and tumor necrosis alpha were expressed. Macrophage migration was also increased. Treatment with ibudilast down-regulated these molecules, suppressed macrophage migration into the aneurysm wall, and inhibited PDE-4 activation and the elevation of cyclic adenosine monophosphate in endothelial cells. These results suggest that blocking of PDE4 is associated with the reduction of inflammation-related molecules and macrophage migration, thereby reducing the progression of cerebral aneurysms. It may represent a new conservative therapy to treat patients with cerebral aneurysms.


Omalizumab in patients with severe asthma and persistent sputum eosinophilia.
Omalizumab, a recombinant humanized monoclonal antibody targeting the IgE molecule, is the first biologic approved for moderate-to-severe allergic asthmatics, who remain uncontrolled despite high dose inhaled corticosteroid and bronchodilators. Steroid-sparing effect of omalizumab has not been demonstrated in asthmatics with persistent airway eosinophilia in a randomised controlled trial till date. From this double-blind, placebo-controlled, multi-centred, randomized parallel group design, we report that omalizumab is possibly inadequate to control sputum eosinophilia, and therefore may not have a steroid-sparing effect, especially in those maintained on oral corticosteroids daily. This needs to be confirmed or refuted in a larger trial, which may be a challenge with respect to recruitment, since there are currently three additional biologics available to prescribe. Trial registration Clinicaltrials.gov, NCT02049294, Registered 30th January 2014, https://clinicaltrials.gov/ct2/show/NCT02049294.


High energy shock waves enhance the cytotoxic effect of doxorubicin and methotrexate to human osteosarcoma cell lines.
Despite recent advances, the prognosis of relapsed osteosarcoma patients remains very poor. Application of high energy shock waves may change the tumour cell growth and increase the cytotoxic effect of in vivo and in vitro chemotherapeutic agents. We studied the effect of their association with doxorubicin or methotrexate on three in vitro osteosarcoma cell lines. The effect of these combinations on SJSA-1, MG-63 and HOS human osteosarcoma cell lines were evaluated through incubation with doxorubicin or methotrexate and high energy shock wave treatment with 1000 shots at 0.22 mJ/mm(2) and an evaluation of the cell number, cell proliferation and apoptosis at days 1, 3 and 6 from the start of treatment. The combination of high energy shock waves and doxorubicin induced a statistically significant advantage in terms of a decrease in cell number on the SJSA-1 and HOS cell lines, a decrease of cell proliferation on all three cell lines and an increase of apoptosis only on the SJSA-1 cell line. The combination of high energy shock waves with methotrexate induced a decrease of the cell number only in the SJSA-1 and in the HOS cell lines, of the cell proliferation in the SJSA-1 and in the MG-63 cell lines, and an increase of apoptosis only on the SJSA-1 cell line. In conclusion, these experiments show an interesting effect of high energy shock waves on osteosarcoma cell lines, which could lead to future studies of the in vivo effects of high energy shock waves in the murine model as well.


Use of vancomycin pharmacokinetic-pharmacodynamic properties in the treatment of MRSA infections.
Vancomycin is a commonly used antimicrobial in patients with methicillin-resistant Staphylococcus aureus (MRSA) infections. Increasing vancomycin MIC values in MRSA clinical isolates makes the optimization of vancomycin dosing pivotal to its continued use. Unfortunately, limited data exist regarding the optimal pharmacokinetic-pharmacodynamic (PK-PD) goal to improve bacterial killing and clinical outcomes with vancomycin. The hallmark study in this area suggests that achieving an AUC to MIC ratio of over 400 improves the likelihood of achieving these outcomes. Challenges in the implementation of PK-PD-based dosing for vancomycin include current methodologies utilized in microbiology laboratories, as well as intra- and interpatient pharmacokinetic variability. Individualized dosing based on MIC and specific patient factors is important to achieve optimal outcomes from vancomycin therapy.


Anxiety and stress in pregnancy: significance and nursing assessment.
This article explores the significance of research on prenatal anxiety and stress for application to clinical nursing practice. The focus will be on assessment of the gravid woman's anxiety levels and coping skills. An introduction presenting a paradigm of the transitional and paradoxical changes of pregnancy precedes a discussion of the nature and sources of anxiety in pregnancy and its effects on maternal/fetal health outcomes. The cumulative effects of multiple sources of anxiety and the relevance of early state anxiety and prenatal personality dimensions, such as maternal role identification and conjugal relationships, to health outcomes also are discussed. Methods for stress reduction and recommendations for future multidisciplinary and multivariate research projects conclude the article.


Idiopathic acquired sideroblastic anaemia transforming to acute myelosclerosis.
A case of idiopathic acquired sideroblastic anaemia transforming to acute myelofibrosis is reported. The appearance of atypical megakaryocytic proliferation in idiopathic acquired sideroblastic anaemia may presage the development of an acute myelofibrotic phase of this usually chronic disease.