Platelet vasopressin levels in childhood idiopathic nephrotic syndrome.
Despite the importance of disturbances in plasma volume in nephrotic patients, the only clinically accepted measurement of this value in common use is plasma renin activity. We assessed the usefulness of plasma vasopressin levels as an index of plasma volume in patients with idiopathic nephrotic syndrome. However, since 80% to 90% of arginine vasopressin circulates bound to platelets, we measured vasopressin levels in platelet-rich and platelet-poor plasma. The nephrotic patients (n = 19) had significantly higher vasopressin levels in platelet-poor and platelet-rich plasma compared with controls (3.2 +/- 0.6 vs 1.0 +/- 0.3 pg/mL, and 10.4 +/- 3.6 vs 3.3 +/- 0.6 pg/mL. respectively). The percent binding of vasopressin to platelets was reduced in nephrotic patients compared with controls (50.2% +/- 6% vs 70.4% +/- 2.9%). The values for platelet-poor vasopressin, but not platelet-rich vasopressin, correlated significantly with the plasma renin activity (r = .83). We conclude that in nephrotic patients, platelet-poor vasopressin levels correlate with plasma renin activity and may provide a useful measure of minute-to-minute vasopressin release in response to changes in plasma volume.


[Contamination of drinking water by nitrates: analysis of health risks].
Nitrate ground water contamination is a relatively frequent problem due to the massive use of fertilizers in agriculture. The health effects of contamination are due to the transformation of nitrates into nitrites in the G.I. tract and possibly the transformation of nitrites into nitrosamines in the stomach. The risk of methemoglobinemia in infants is due to nitrites contained in the water used to reconstitute milk for feeding. There are possible but unproven risks associated with the formation of nitrosamines, i.e. teratogenous and cancer risks. Drinking water standards are only based on the risk of methemoglobinemia, but given the present state of knowledge they appear to be safer in relation to the other potential risks. In the face of the increase in nitrates in ground water and the partially known risks, it appears necessary to try and prevent this type of contamination and inform the persons at risk, such as pregnant women and young mothers. It would also be useful to promote research on the effects of nitrates and nitrites on humans.


[Nursing process in advanced cardiopulmonary resuscitation].
The process male nurse is a systematic and organized method to offer effective and efficient cares guided to the achievement of solving real problems of health, reducing the incidence and the duration. It is organized and systematic for that consists of five sequential and interrelated steps: Valuation, diagnostic, planning, execution and evaluation, in which are carried out interrelated actions, thought to maximize the long term results. The nurse process is based on the notion that the success of the cares is measured by the degree of effectiveness and the degree of satisfaction and the patient's progress. Applying this method in the Advanced Cardiac Live Support (ACLS) the identification of a cardiovascular or cardiopulmonary urgency was achieved that implies advanced treatment of the air road, defibrillation and appropriate medications to the circumstances. The ACLS challenges the nurses in charge from the patient's attention to make decisions quick low pressure and in dramatic scenes. Reason why it develops the flowing process male nurse in the advanced cardiopulmonary reanimation due to the incidence of these events in the National Institute of Cardiology Ignacio Chávez, which should guarantee the benefit of services in basic and advanced cardiopulmonary reanimation for personal with a high formation level in all the units of intensive cares and services of hospitalization in integrated form and stratified this way to avoid that it progresses to situations that cause the death or leave irreversible sequels since in the central nervous system the time it is a factor critical for the treatment of this events.


PET/CT-guided treatment planning for paediatric cancer patients: a simulation study of proton and conventional photon therapy.
To investigate the impact of including fluorine-18 fludeoxyglucose ((18)F-FDG) positron emission tomography (PET) scanning in the planning of paediatric radiotherapy (RT). Target volumes were first delineated without and subsequently re-delineated with access to (18)F-FDG PET scan information, on duplicate CT sets. RT plans were generated for three-dimensional conformal photon RT (3DCRT) and intensity-modulated proton therapy (IMPT). The results were evaluated by comparison of target volumes, target dose coverage parameters, normal tissue complication probability (NTCP) and estimated risk of secondary cancer (SC). Considerable deviations between CT- and PET/CT-guided target volumes were seen in 3 out of the 11 patients studied. However, averaging over the whole cohort, CT or PET/CT guidance introduced no significant difference in the shape or size of the target volumes, target dose coverage, irradiated volumes, estimated NTCP or SC risk, neither for IMPT nor 3DCRT. Our results imply that the inclusion of PET/CT scans in the RT planning process could have considerable impact for individual patients. There were no general trends of increasing or decreasing irradiated volumes, suggesting that the long-term morbidity of RT in childhood would on average remain largely unaffected. (18)F-FDG PET-based RT planning does not systematically change NTCP or SC risk for paediatric cancer patients compared with CT only. 3 out of 11 patients had a distinct change of target volumes when PET-guided planning was introduced. Dice and mismatch metrics are not sufficient to assess the consequences of target volume differences in the context of RT.


Interictal EEG, hippocampal atrophy, and cell densities in hippocampal sclerosis and hippocampal sclerosis associated with microscopic cortical dysplasia.
The EEG characteristics of isolated hippocampal sclerosis (HS) and HS associated with other types of temporal lobe pathology are not well defined. The pathologic substrate may be an important variable in determining seizure-free outcome. The objective of this study was to define the distribution of epileptiform discharges in patients with HS and HS associated with microscopic dysplasia, and to examine their relationship with hippocampal atrophy and cell loss. Thirty-four patients (15 women and 19 men; mean age, 30.6 +/- 11.2 years), all with good outcomes after temporal lobectomy (Engel classes I and II), were included. The characteristics studied were frequency and distribution of spikes, MRI-based hippocampal volume ratios, and quantitative hippocampal cell density in various subregions. The isolated HS group showed a trend to a higher percentage of epileptiform discharges maximal at the anterior temporal electrodes (89.87 +/- 17.0%; 79.5 +/- 28.2% in the dual-pathology group). The isolated HS group had, on average, significantly more cell loss (P < 0.001). There was a significant negative correlation between the amount of cell loss in the CA1 area and both anterior temporal spikes and hippocampal ratios (P < 0.05). Isolated HS and dual pathology show minimal differences in interictal spike distribution and frequency. More widespread spike distributions in severe isolated HS compared with patients with less cell loss is probably the result of less organized limbic circuitry.


[Contralateral local anesthesia in stump, phantom and post-traumatic pain].
Contralateral local anaesthesia was employed in patients with chronic (n = 42) and acute (n = 8) pain syndromes (see Table 1). I. Phantom limb and stump pain: n = 10; II. Pain of face and ear: n = 10; III. Pain of neck and trunk: n = 14; IV. Pain of the hip joint: n = 6; V. Pain of the extremities: n = 10: Sa. = 50 patients. Among these were n acute pain syndromes: I.: n = 0; II.: n = 1; III.: n = 1; IV.: n = 1; V.: n = 5 (Sa.: n = 8). 42 patients with chronic pain syndromes were treated with contralateral local anaesthesia (CLA). In 27 patients CLA displayed a clear effect upon the chronic pain; in 8 patients pain release was more than 50%, in 15 patients pain release was less than 50%. 3 patients relapsed into their former condition of pain. CLA was without any positive influence in 12 cases. The medium frequency of treatments was between 5 and 6 in both groups. The average time of treatment amounted to 6 and 7 months respectively. In 8 patients with acute pain syndromes contralateral local anaesthesia produced the following results: 6 patients were permanently released from their pain; in 2 patients CLA remained without any success. The average frequency of treatment was 1.25; the average time of observation was 2.3 months. Hence we suggest that CLA should be employed as early as possible in acute or chronic posttraumatic or postoperative pain syndromes. The influence of contralateral pain therapy on acute and chronic pain conditions of the opposite side can be affirmed; its mechanism remains to be clarified. Obviously it is possible to exert an inhibitory influence from the contralateral side upon peripheral, spinal, reticular and thalamic regions, which can lead to the extinction of acute and chronic pain conditions on the opposite side. According to our experiences it is necessary that the cerebro-spinal nerve system - apart from the traumatic lesion - is intact and that the pain syndrome is not maintained by a psychic disturbance.


Incipient periodontitis and salivary molecules among Korean adults: association and screening ability.
This study aims to evaluate the association of salivary matrix metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8) with periodontitis and its screening ability on periodontitis. We selected 693 participants from the Yanypyeong Cohort: 577 participants with periodontitis and 116 participants without periodontitis. Periodontitis was assessed by dentists using a panoramic radiograph. Salivary MMP-9 and IL-8 were assayed using multiplexed bead immunoassay (Luminex). MMP-9 and IL-8 were categorized into low, medium and high. Age, sex, income, smoking, drinking, exercise, obesity and metabolic syndrome were confounders. Logistic regression analysis was applied to evaluate the association between MMP-9, IL-8 and periodontitis. Receiver operating characteristic curve was applied for sensitivity, specificity and c-statistics. High MMP-9 and medium IL-8 were associated with periodontitis: adjusted odds ratio were 2.2 [95% confidence interval (CI): 1.3-3.7] for MMP-9 and 1.9 (95% CI: 1.1-3.4) for IL-8. The final screening model using salivary MMP-9 for periodontitis had a sensitivity of 0.46, specificity of 0.77 and c-statistic of 0.63 (p < 0.001). Our data show that salivary MMP-9 and IL-8 could be potential markers for periodontitis. The screening model for periodontitis could be useful in clinics and home. A future prospective study is indicated for predicting the occurrence of periodontitis.


Total occlusion of the abdominal aorta in a patient with renal failure and refractory hypertension: a case report.
Total occlusion of the abdominal aorta is unusual, and potentially catastrophic. It occurs in patients with advanced atherosclerotic occlusive disease, and can cause severe ischemic manifestations, depending on the site of obstruction. Prompt and appropriate diagnostic and therapeutic approaches are important whenever this condition is suspected, in order to avoid a fatal outcome. The development of a complex network of collaterals may prevent the manifestation of acute ischemic phenomena, and cause a delay in diagnosis and treatment. Here we report the clinical case of a 59-year-old man who was referred to our Department for evaluation of renal failure and refractory hypertension. Ultrasonography and 99mTc-DTPA scintigraphy showed a shrunken, non-functioning left kidney, while CT angiography and aortography showed the complete occlusion of the aorta from below the right renal artery down to the bifurcation of both common iliac arteries, with a critical stenosis of the origin of the right renal artery, an occlusion of the left renal artery as well as of the origin of the inferior mesenteric artery. The patient was referred to the surgery department for aorto-bifemoral bypass surgery and re-implantation of the right renal artery.


The mechanism of chick blastoderm expansion.
At the time of laying, the domestic fowl blastoderm measures 4 mm across. After 4 days' incubation, the extra-embryonic yolk-sac tissues have expanded to encompass the whole yolk mass. This expansion involves the migration over the inner surface of the vitelline membrane of a specialized band of 'edge cells' at the blastoderm periphery. As they move, they pull out the blastoderm behind them, setting up a considerable tension. Expansion also involves cell proliferation and changes in cell shape. This paper attempts to show how locomotion, tension, proliferation and changes in cell shape all contribute to the orderly process of expansion. As a simplification, only the extra-embryonic epiblast is considered here. The findings are: 1. Expansion does not occur at a constant rate, but starts slowly, rises to a peak (over 500 mum/h) at around 3 days, and then slows as coverage of the yolk mass nears completion. 2. During the first day of incubation, edge-cell migration produces a tension in the blastoderm. This rises to peak at 20-24 h, then declines. This tension may be due to an imbalance between expansion by migration and expansion by proliferation. 3. Migration of edge cells can be affected by tension in the blastoderm, i.e. very high tension may hold them back. However, the tension level normally found in the blastoderm seems not to do so. The low rate of expansion in the first day is therefore not due to the high level of tension. It may instead be due to changes in edge-cell organization. 4. Proliferation occurs throughout the extra-embryonic epiblast during the expansion period. It is not restricted to the blastoderm periphery. After the yolk has been covered, the epiblast continues to grow, with proliferation restricted largely to band just distal to the advancing edge of the area vasculosa. 5. Cell shape and arrangement change considerably during expansion. The epiblast of the unincubated embryo is a monolayer of tall cells. During expansion, these become considerably flattened so that each contributes a larger amount to yolk-sac surface area.


Analysis of influenza A H3N2 strains isolated in England during 1995-1996 using polymerase chain reaction restriction.
A polymerase chain reaction-restriction (PCR-restriction) endonuclease assay was developed to allow rapid analysis of influenza A H3N2 viruses circulating in England during 1995-1996. Restriction endonuclease digestion with two enzymes of amplicons derived from PCR of the HA1 portion of the influenza haemagglutinin (HA) gene was able to differentiate antigenically similar influenza strains into two groups. Group I variants were similar genetically to the 1995/96 vaccine strain, A/Johannesburg/33/94, whereas the HA sequences of Group II variants were similar genetically to the reference virus A/Thessaloniki/1/95. Of the 700 England A H3N2 strains isolated between February 1995 and the end of April 1996, 384 were analysed by this method. PCR-restriction analysis of sequential influenza isolates revealed a temporal alteration in prevalence of two variants. Groups I and II variants cocirculated with equal frequency during a period of sporadic influenza activity, but following the onset of epidemic influenza activity in 1995, only Group II variants were detected. PCR- restriction analysis was found to be a rapid method for studying genetic variation which could be applied to a large number of samples and provide information about the direction of genetic drift in the HA gene of influenza virus.


Heterogeneous lymphokine-activated killer cell precursor populations. Development of a monoclonal antibody that separates two populations of precursors with distinct culture requirements and separate target-recognition repertoires.
We developed a monoclonal antibody (mAb) 211, which recognizes the precursors in peripheral blood of lymphokine-activated killer cells (LAK) induced by recombinant interleukin-2 (rIL-2). In conjunction with complement mAb 211 also eliminates natural killer cells (NK) and a majority of the cytotoxic T lymphocytes. B cells and monocytes do not express the 211 antigen. Since mAb 211 recognized such a large percentage of peripheral blood lymphocytes we examined which 211+ subpopulation was the predominant precursor of rIL-2-induced LAK cells using two-color fluoresence-activated cell sorting (fluorescein-conjugated 211 mAb plus phycoerythrin-CD11b). This method identified the 211+/CD11b+ population as the predominant phenotype of the rIL-2-induced LAK precursor. In addition, we directly compared the phenotype of the LAK precursor induced by delectinated T-cell growth factor (TCGF) to that induced by rIL-2. The 211-depleted population, which was devoid of NK cells and LAK precursors (inducible by rIL-2), was capable of generating LAK activity when TCGF was used as the source of lymphokine. LAK cells induced by TCGF from the 211-depleted population lysed a fresh sarcoma and an NK-resistant cultured melanoma tumor target but not the Daudi cell line, which was lysed by rIL-2-induced LAK cells. Lymphoid subpopulations, depleted using NKH1a mAb, behaved similarly, generating high levels of lysis against the two solid tumor targets when cultured with TCGF but not with rIL-2. CD 3-depleted populations showed enrichment for LAK precursors using either rIL-2 or TCGF. These results indicate that while rIL-2-induced LAK precursors cannot be separated from cells with NK activity, TCGF-induced LAK cells can be generated from populations of peripheral blood mononuclear cells without NK activity.


Error of finite basis expansions in time-dependent calculations of atom-laser interaction.
We propose a method to quantitatively estimate the error made with a finite basis expansion in time-dependent calculations. This method is applied to the hydrogen atom in intense laser fields and used to compare different basis sets with each other. We also show how to construct a Gaussian basis set suitable for the description of ionization dynamics in intense laser fields.


Aestivation Induces Changes in the mRNA Expression Levels and Protein Abundance of Two Isoforms of Urea Transporters in the Gills of the African Lungfish, Protopterus annectens.
The African lungfish, Protopterus annectens, is ammonotelic in water despite being ureogenic. When it aestivates in mucus cocoon on land, ammonia is detoxified to urea. During the maintenance phase of aestivation, urea accumulates in the body, which is subsequently excreted upon arousal. Urea excretion involves urea transporters (UT/Ut). This study aimed to clone and sequence the ut isoforms from the gills of P. annectens, and to test the hypothesis that the mRNA and/or protein expression levels of ut/Ut isoforms could vary in the gills of P. annectens during the induction, maintenance, and arousal phases of aestivation. Two isoforms of ut, ut-a2a and ut-a2b, were obtained from the gills of P. annectens. ut-a2a consisted of 1227 bp and coded for 408 amino acids with an estimated molecular mass of 44.7 kDa, while ut-a2b consisted of 1392 bp and coded for 464 amino acids with an estimated molecular mass of 51.2 kDa. Ut-a2a and Ut-a2b of P. annectens had a closer phylogenetic relationship with Ut/UT of tetrapods than Ut of fishes. While the mRNA expression pattern of ut-a2a and ut-a2b across various tissues of P. annectens differed, the transcript levels of ut-a2a and ut-a2b in the gills were comparable, indicating that they might be equally important for branchial urea excretion during the initial arousal phase of aestivation. During the maintenance phase of aestivation, the transcript level of ut-a2a increased significantly, but the protein abundance of Ut-a2a remained unchanged in the gills of P. annectens. This could be an adaptive feature to prepare for an increase in the production of Ut-a2a upon arousal. Indeed, arousal led to a significant increase in the branchial Ut-a2a protein abundance. Although the transcript level of ut-a2b remained unchanged, there were significant increases in the protein abundance of Ut-a2b in the gills of P. annectens throughout the three phases of aestivation. The increase in the protein abundance of Ut-a2b during the maintenance phase could also be an adaptive feature to prepare for efficient urea excretion when water becomes available.


Scanning electron microscopy of the jejunum in children with protein-energy malnutrition.
Samples of jejunum from seven children dying from severe oedematous malnutrition were fixed with intralumenal glutaraldehyde in combination with external immersion, using a rapid autopsy protocol. Selected areas were post-fixed in osmium tetroxide, dehydrated with an ethanol series and critical point dried. After mounting on aluminium stubs, specimens were sputter-coated with a thin layer of gold and palladium. Scanning electron microscopy showed patterns ranging from subtotal villous atrophy, through partial villous atrophy with low ridges, to longer, taller ridges with cerebriform convolutions. In one case, villous height had been interpreted as normal on light microscopy. The scanning electron microscope revealed that the mucosa in this case was composed of ridges and leaf forms. Processing for scanning electron microscopy is simple. The specimens can be viewed and photographed in a relatively short time. In addition, the three-dimensional record provides clarification of ambiguities which often arise from routine histological sections.


Five-Year Survival is Not a Useful Measure for Cancer Control in the Population: an Analysis Based on UK Data
Background: Five-year survival is an important metric for progress in cancer control broadly used both in the cancer literature and by the public. In order to assess its validity and relation to other common metrics, we analyzed the relationship between 5-year survival, incidence and mortality using publicly available cancer registry data from England and Wales. Methods: Five-year survival, incidence and mortality data were obtained from the online database of a registered charity, Cancer Research UK. We extracted sex-specific age-standardized mortality, incidence, and 5-year survivalfor16 types of cancer over the period from 1976 to 1995. The relationships between 5-year survival, incidence and mortality were estimated using both Pearson and Spearman correlation coefficients. Results: All 16cancer types showed an increase in 5-year survival for both genders from 1976 to 1995, ranging from 0.2% (pancreas and lung cancer) to 16.6% (prostate cancer) for males and 0.2% (pancreas cancer) to 16.6% (leukemia) for females. From 1976 to 1995, there was no significant correlation between changes in 5-year survival and cancer mortality for either sex (males, Pearson r=0.16, Spearman r=-0.06; females, Pearson r=-0.33, Spearman r=-0.43). A positive relationship between 5-year survival and incidence was noted among males, but not among females (males, Pearson r=0.61, Spearman r=0.53; females, Pearson r=0.03, Spearman r=0.11). However, after excluding breast and prostate cancer, the positive association became weaker and became statistically non-significant for males (Pearson r=0.47; Spearman r=0.41). Conclusions: Our findings suggest that there are no reliable relationships between changes in 5-year survival and cancer incidence or mortality. Increases in 5-year survival might therefore represent poor indicators of progress in cancer control at the population level. In the absence of over-diagnosis, 5-year survival might only indicate improved diagnosis and treatment in clinical practice.


Use of amphotropic retroviral vectors for gene transfer in human colon carcinoma cells.
Previous studies in rodent models have demonstrated the feasibility of gene transfer to the stem cells of the intestinal epithelium using ecotropic retroviral vectors delivered luminally. This report represents a next step toward targeting the human intestine as a site for somatic gene therapy. The first experiment assessed the viability of amphotropic retroviral vectors in the luminal environment. It was found that after 4 hr at 37 degrees C in luminal effluent, the loss of titer was no greater than when incubated in control media. Likewise, neither the vector nor the target cells were adversely affected by N-acetylcysteine, which is likely to be used as a preparatory agent for mucus removal. To determine whether human intestinal cells are transducible by these vectors, three colon carcinoma cell lines were studied: HT-29, T84, and Caco-2. All were transduced; however, the expression of the reporter gene was highest in the HT-29 cells. Subsequent studies using these cells showed that with regular stocks of vector, gene transfer peaked at a stock dilution of 1/10 and declined at full strength. This problem could be partially overcome by centrifugal concentration of the retroviral stocks. With this approach, gene transfer increased with increasing particles up to 10x regular stock titers but was inefficient at 100x. Overall, these findings provide encouraging evidence that amphotropic retroviral vectors may eventually be used for in vivo gene transfer into human intestinal epithelium. However, they also point to the need for improved methods of concentrating retroviral vectors.


Compendium of naringenin: potential sources, analytical aspects, chemistry, nutraceutical potentials and pharmacological profile.
Naringenin is flavorless, water insoluble active principle belonging to flavanone subclass. It exhibits a diverse pharmacological profile as well as divine nutraceutical values. Although several researchers have explored this phytoconstituent to evaluate its promising properties, still it has not gained recognition at therapeutic levels and more clinical investigations are still required. Also the neutraceutical potential has limited marketed formulations. This compilation includes the description of reported therapeutic potentials of naringenin in variety of pathological conditions alongwith the underlying mechanisms. Details of various analytical investigations carried on this molecule have been provided along with brief description of chemistry and structural activity relationship. In the end, various patents filed and clinical trial data has been provided. Naringenin has revealed promising pharmacological activities including cardiovascular diseases, neuroprotection, anti-diabetic, anticancer, antimicrobial, antiviral, antioxidant, anti-inflammatory and anti-platelet activity. It has been marketed in the form of nanoformulations, co-crystals, solid dispersions, tablets, capsules and inclusion complexes. It is also available in various herbal formulations as nutraceutical supplement. There are some pharmacokinetic issue with naringenin like poor absorption and low dissolution rate. Although these issues have been sorted out upto certain extent still further research to investigate the bioavailability of naringenin from herbal supplements and its clinical efficacy is essential.


Dexamethasone increases adrenalectomy-depressed Na+,K(+)-ATPase mRNA and ouabain binding in spinal cord ventral horn.
The Na+,K(+)-ATPase plays a key role in the regulation of ion fluxes and membrane repolarization in the CNS. We have studied glucocorticoid effects on biosynthesis of the Na+,K(+)-ATPase and on ouabain binding in the ventral horn of the spinal cord using intact rats, adrenalectomized (ADX) rats, and ADX rats receiving dexamethasone (ADX+DEX) during 4 days. Cryostat sections from spinal cords were incubated with a 35S-oligonucleotide coding for the alpha 3-subunit or a 3H-cDNA coding for the beta 1-subunit of the Na+,K(+)-ATPase using in situ hybridization techniques. In ventral horn motoneurons, grain density per cell and grain density per area of soma for both probes were slightly reduced in ADX rats but significantly increased in the ADX+DEX group, using ANOVA and the Bonferroni's test. Statistical analysis of frequency histograms of neuronal densities further indicated a significant shift to the right for intact rats compared with ADX rats for both probes. Concomitantly, [3H]ouabain binding to membrane preparations from ventral horns was reduced in ADX rats and restored to normal by DEX administration. No effect of adrenalectomy or DEX treatment was obtained in the dorsal horn. In conclusion, glucocorticoids positively modulate the mRNA for the alpha 3-subunit and the beta 1-subunit of the Na+,K(+)-ATPase and recover ouabain binding to normal values. The increments of the synthesis and activity of an enzyme affecting membrane repolarization and synaptic neurotransmission are consistent with the alleged stimulatory effect of glucocorticoids on spinal cord function.


The metagenomics of soil bacteria and fungi and the release of mechanical dormancy in hard seeds.
Persistence in the soil is a function of seed physiology, particularly non-germination and inherent lifespan. However, for seeds with mechanical dormancy, non-germination is also a function of the composition and activity of the soil microbiota. We attempted to screen out microorganisms in the soil that can specifically and rapidly decompose the hard fruit pericarps of Tilia miqueliana Maxim., a unique native tree species in China. Using the classical replica plating method, more than 100 different culturable microorganisms that could rapidly erode the pericarp were collected from the surface of pericarps under different culture conditions. At the same time, we successfully extended the concept of metagenomics and applied it to the identification of mixed artificial cultures. The decomposition process of the pericarps in soil was also simulated artificially. The physical and chemical data suggested a potential mechanism of microbial scarification and cracking in pericarp, whilst the embryos inside the eroded fruits retained good viability. Our discoveries could pave the way for the removal of physical and mechanical obstacles that prevent hard coat seeds from germinating. We anticipate that the use of this technology will improve the germination of other hard coat seeds. More research is needed to investigate the impacts on other seeds. The findings of this research can inform the design of experiments on the seed ecology of persistence.


The bipartite architecture of the sRNA in an archaeal box C/D complex is a primary determinant of specificity.
The archaeal box C/D sRNP, the enzyme responsible for 2'-O-methylation of rRNA and tRNA, possesses a nearly perfect axis of symmetry and bipartite structure. This RNP contains two platforms for the assembly of protein factors, the C/D and C'/D' motifs, acting in conjunction with two guide sequences to direct methylation of a specific 2'-hydroxyl group in a target RNA. While this suggests that a functional asymmetric single-site complex complete with guide sequence and a single box C/D motif should be possible, previous work has demonstrated such constructs are not viable. To understand the basis for a bipartite RNP, we have designed and assayed the activity and specificity of a series of synthetic RNPs that represent a systematic reduction of the wild-type RNP to a fully single-site enzyme. This reduced RNP is active and exhibits all of the characteristics of wild-type box C/D RNPs except it is nonspecific with respect to the site of 2'-O-methylation. Our results demonstrate that protein-protein crosstalk through Nop5p dimerization is not required, but that architecture plays a crucial role in directing methylation activity with both C/D and C'/D' motifs being required for specificity.


Febuxostat for management of tumor lysis syndrome including its effects on levels of purine metabolites in patients with hematological malignancies - a single institution's, pharmacokinetic and pilot prospective study.
Tumor lysis syndrome (TLS) is a life-threatening oncological emergency, and control of serum uric acid level (S-UA) is most important. In this single-institution, short-term and pilot prospective study, the efficacy of a new xanthine oxidase inhibitor, febuxostat, as an alternative to conventional allopurinol, including its effects on hypoxanthine and xanthine, was evaluated in 10 consecutive patients with hematological malignancies at intermediate risk for TLS. Febuxostat at 40 mg (n=7) or 60 mg (n=3) daily was administered according to renal function, and induction chemotherapy was started within 24 h. The primary end-point was the reduction of S-UA to ≤ 7.5 mg/dl by day 5. The median S-UA at base-line was 8.0 mg/dl (range=3.2-10.6 mg/dl). The median S-UA on day 5 after chemotherapy was 3.3 mg/dl (range=1.1-5.8 mg/dl) (p<0.0001, by paired t-test), indicating successful control of S-UA during chemotherapy. All patients achieved S-UA ≤ 7.5 mg/dl. A simultaneous decrease in serum creatinine and increase in estimated glomerular filtration rate were seen. Serum hypoxanthine and xanthine levels (as the consequence of inhibition of xanthine oxidase) were elevated along with the decrease in S-UA. Xanthine level was elevated higher compared to hypoxanthine level and reached the level reported to cause xanthine nephropathy, but no advance of renal impairment was observed. Serum febuxostat concentrations at 2 h after administration were 891.8 ± 285.0 ng/ml (mean ± SE) for the 40-mg dose and 770.6 ± 242.7 ng/ml for the 60-mg dose (p=0.80, unpaired t-test), showing no accumulation in patients with renal impairment. No febuxostat-related adverse reactions were noted. No patients experienced progressive TLS. Febuxostat is promising for the management of TLS of an intermediate-risk patient and further observation and reevaluation regarding xanthine nephropathy should be performed.


Clinical course of sub-centimeter-sized nodules detected during surveillance for hepatocellular carcinoma.
To evaluate the outcome of sub-centimeter-sized nodules (SCSNs) detected during surveillance for hepatocellular carcinoma (HCC) in patients at risk. We retrospectively analyzed a total of 142 patients with liver cirrhosis or chronic hepatitis B or C without a prior history of HCC in whom a SCSN was detected during HCC surveillance. We calculated the rate of HCC development from SCSNs in the study population and analyzed the differences in the baseline clinical characteristics and imaging features between the patients with SCSNs that eventually developed into HCC and patients with SCSNs that did not develop into HCC. During 667 person-years of follow-up, HCC developed in 33 patients. The calculated HCC development rate was 4.9% per year. The cumulative one-, two-, three- and five-year HCC development rates were 5.6%, 10.6%, 14.1% and 20.4%, respectively. Upon baseline comparison, the HCC group was older (54.4 ± 8.3 years vs 48.9 ± 9.4 years; P = 0.003) and had lower albumin levels (3.56 ± 0.58 g/dL vs 3.84 ± 0.55 g/dL; P = 0.012) and higher baseline alpha-fetoprotein (AFP) levels (8.5 ng/mL vs 5.4 ng/mL; P = 0.035) compared to the non-HCC group. Nodule pattern and initial radiologic diagnosis also differed between the two groups. Multivariate analysis revealed that age [P = 0.012, odds ratio (OR) =1.075, 95% confidence interval (CI) =1.016-1.137], sex (P = 0.009, OR = 3.969, 95% CI: 1.403-11.226), and baseline AFP level (P = 0.024, OR = 1.039, 95% CI: 1.005-1.073) were independent risk factors for developing HCC. The overall risk of HCC development in patients with SCSNs is similar to that in liver cirrhosis patients. Patients with these risk factors need to be closely monitored during follow-up.


Pharmacodynamics of long-acting folic acid-receptor targeted ritonavir-boosted atazanavir nanoformulations.
Long-acting nanoformulated antiretroviral therapy (nanoART) that targets monocyte-macrophages could improve the drug's half-life and protein-binding capacities while facilitating cell and tissue depots. To this end, ART nanoparticles that target the folic acid (FA) receptor and permit cell-based drug depots were examined using pharmacokinetic and pharmacodynamic (PD) tests. FA receptor-targeted poloxamer 407 nanocrystals, containing ritonavir-boosted atazanavir (ATV/r), significantly increased drug bioavailability and PD by five and 100 times, respectively. Drug particles administered to human peripheral blood lymphocyte reconstituted NOD.Cg-Prkdc(scid)Il2rg(tm1Wjl)/SzJ mice and infected with HIV-1ADA led to ATV/r drug concentrations that paralleled FA receptor beta staining in both the macrophage-rich parafollicular areas of spleen and lymph nodes. Drug levels were higher in these tissues than what could be achieved by either native drug or untargeted nanoART particles. The data also mirrored potent reductions in viral loads, tissue viral RNA and numbers of HIV-1p24+ cells in infected and treated animals. We conclude that FA-P407 coating of ART nanoparticles readily facilitates drug carriage and antiretroviral responses.


[Somatomedin (bioassay) response to long-term growth hormone treatment of idiopathic pituitary dwarfism].
Serum somatomedin (SM) activity by bioassay was measured in 46 idiopathic pituitary dwarfs before and during long-term hGH treatment. The mean (+/- SD) pretreatment SM activity was 0.38 +/- 0.17 U/ml. The mean (+/- SD) pretreatment annual growth rate was 3.4 +/- 1.0 cm/year. None of the patients had a SM activity within the normal range of age-matched controls. The mean SM activity 1 day post GH was 1.11 +/- 0.28 U/ml. Neither the mean SM activity nor the growth rate during the entire treatment period up to 8 years decreased with the duration of the treatment, always being more than 1.0 U/ml and 5.0 cm/year respectively. With respect to normal data for chronological and bone age separately, 43.3% and 60% of the SM activity in the patients during hGH treatment attained the levels of age-matched normal controls. The increase in SM activity in the patients with treatment was not related to their growth rate. SM activity measured by bioassay is useful for evaluating clinical response to hGH therapy.


A parsimonious scoring and normative calculator for the Parkinson's disease mild cognitive impairment battery.
The aim of the present study was to provide a regression-based calculator that takes premorbid functioning into account to detect subtle cognitive decline, as is often present in pre-dementia states, especially mild cognitive impairment in Parkinson's disease (PD-MCI). We used demographic adjustments based on sex, age, and education of 699 normative participants that fulfilled exclusion criteria for ascertaining the diagnostic accuracy of the Movement Disorders Society PD-MCI battery at Level II. We examined the clinical validity of the battery on 36 PD patients. An estimated z-score was calculated for any raw score based on different models that adjust for the demographic predictors of gender, age, and education, either concurrently, individually or without covariates. We provide a useful online z-score, SD, and percentile calculator that yields estimates of cognitive impairment based on normative sample for each of the ten neuropsychological tests and enables actuarial decision-making regarding its level and profile (number of domains impaired). We document the clinical utility and applicability of the calculator on a patient with PD-MCI and show the discriminative validity of all measures in the battery by comparing PD-MCI and PD without cognitive impairment with the highest area under the curve (.94) for Tower of London at -2 SD threshold. Our normative calculator introduces a practical web-based psychometric tool for the evaluation of PD-MCI status in clinical settings. We show the detection potential of each of ten measures included in the battery and delineate the diagnostic precision of the PD-MCI battery.


My Phenotype speaks: please do not harm me with biopsy needle.
Fibrodysplasia ossificans progressiva is characterized by congenital skeletal anomalies and progressive heterotopic ossification. We present a 4 year old male patient who underwent unnecessary harmful multiple biopsies before the diagnosis of fibrodysplasia ossificans progressiva is made. Though rare, diagnosis of fibrodysplasia ossificans progressiva should be considered whenever characteristic radiographic features of multifocal heterotopic bone formation is seen along with the valgus deformities of the big toes.


Risk of neoplastic progression in Barrett's esophagus diagnosed as indefinite for dysplasia: a nationwide cohort study.
A histological diagnosis of "indefinite for dysplasia" (IND) in Barrett's esophagus is used when a diagnosis of genuine dysplasia is equivocal. The aim of the present study was to assess the risk of progression to high grade dysplasia (HGD) or esophageal adenocarcinoma (EAC) after a diagnosis of IND in a nationwide cohort of patients with Barrett's esophagus. Patients with a first diagnosis of IND in Barrett's esophagus between 2002 and 2011 were selected from a nationwide registry of histopathology diagnoses in The Netherlands. Patients were followed up until treatment for HGD, detection of EAC, or date of last endoscopy contact with biopsy sampling. In total, 1258 patients met the inclusion criteria, of whom 842 (66.9 %) underwent endoscopic follow-up. Patients were followed for a total of 2585 person-years (mean ± SD 3.01 ± 2.6). Median duration until first follow-up endoscopy was 1.2 years (interquartile range 0.3 - 1.8 years). The progression rate from IND to the combined end point of HGD or EAC was 2.0 (95 % confidence interval [CI] 1.5 - 2.6) per 100 person-years and progression to EAC was 1.2 (95 %CI 0.8 - 1.6). After excluding cases with HGD or EAC within 1 year after IND diagnosis (n = 16), the progression rates were 1.4 (95 %CI 1.0 - 1.9) and 0.8 (95 %CI 0.5 - 1.2) per 100 person-years for HGD or EAC and EAC, respectively. In this large, population-based, cohort of patients with Barrett's esophagus, the incidence rate of HGD or EAC following a diagnosis of IND was 1.4 per 100 person-years. The results demonstrate the need for additional studies to select the subgroup of IND patients with an increased risk of neoplastic progression.


Radixin knockdown by RNA interference suppresses human glioblastoma cell growth in vitro and in vivo.
Radixin, a member of the ERM (ezrin-radixin-moesin) family, plays important roles in cell motility, invasion and tumor progression. It is expressed in a variety of normal and neoplastic cells, including many types of epithelial and lymphoid examples. However, its function in glioblastomas remains elusive. Thus, in this study, radixin gene expression was first examined in the glioblastoma cells, then suppressed with a lentivirus-mediated short-hairpin RNA (shRNA) method.We found that there were high levels of radixin expression in glioblastoma U251cells. Radixin shRNA caused down-regulation of radixin gene expression and when radixin-silenced cells were implanted into nude mice, tumor growth was significantly inhibited as compared to blank control cells or non- sense shRNA cells. In addition, microvessel density in the tumors was significantly reduced. Thrombospondin-1 (TSP-1) and E-cadherin were up-regulated in radixin- suppressed glioblastoma U251 cells. In contrast, MMP9 was down-regulated. Taken together, our findings suggest that radixin is involved in GBM cell migration and invasion, and implicate TSP-1, E-cadherin and MMP9 as metastasis-inducing factors.


Measurement of the membrane potential generated by complex I in submitochondrial particles.
To investigate the energy-conserving function of the NADH:ubiquinone reductase (complex I), we have selected oxonol VI [bis(3-propyl-5-oxoisoxazol-4-yl)pentamethine oxonol] as the most sensitive probe for measuring the reactions of membrane potential generation in submitochondrial particles. Calibration of the oxonol signals with potassium diffusion potentials shows a non-linear response after a threshold around -50 mV. Thermodynamic evaluations indicate that the upper limit of the oxonol response to the potential generated by complex I is around -220 mV, which is close to the maximal protonmotive force in coupled submitochondrial particles. NADH addition to particles in which ubiquinol oxidation is blocked by inhibitors of other respiratory complexes generates oxonol signals corresponding to membrane potentials of -130 to -180 mV. These signals are produced by about four turnovers of the complex reducing endogenous ubiquinone (i.e. non-steady-state conditions) and are equivalent to a charge separation similar to that of the antimycin-sensitive reactions of ubiquinol:cytochrome c reductase (complex III). The transient oxonol signals under non-steady-state conditions are thus informative of crucial steps in the electrogenic reactions catalyzed by complex I. The possible nature of these electrogenic reactions is discussed in relation to proposed mechanisms for complex I.


Love is blind: indiscriminate female mating responses to male courtship pheromones in newts (Salamandridae).
Internal fertilization without copulation or prolonged physical contact is a rare reproductive mode among vertebrates. In many newts (Salamandridae), the male deposits a spermatophore on the substrate in the water, which the female subsequently takes up with her cloaca. Because such an insemination requires intense coordination of both sexes, male newts have evolved a courtship display, essentially consisting of sending pheromones under water by tail-fanning towards their potential partner. Behavioral experiments until now mostly focused on an attractant function, i.e. showing that olfactory cues are able to bring both sexes together. However, since males start their display only after an initial contact phase, courtship pheromones are expected to have an alternative function. Here we developed a series of intraspecific and interspecific two-female experiments with alpine newt (Ichthyosaura alpestris) and palmate newt (Lissotriton helveticus) females, comparing behavior in male courtship water and control water. We show that male olfactory cues emitted during tail-fanning are pheromones that can induce all typical features of natural female mating behavior. Interestingly, females exposed to male pheromones of their own species show indiscriminate mating responses to conspecific and heterospecific females, indicating that visual cues are subordinate to olfactory cues during courtship.


Correlation between nutritional markers and appetite self-assessments in hemodialysis patients.
Protein-energy malnutrition is among the comorbidities that most strongly affect the prognosis of patients with chronic kidney disease. Anorexia, defined as a loss of desire to eat, is one cause of such malnutrition. Tools that evaluate appetite and the correlation between appetite and nutritional parameters require further study. To evaluate the appetite status in patients from 2 hemodialysis clinics in Fortaleza, Brazil and the correlations between appetite (evaluated in the past week and in the past 4 weeks) and demographic, laboratory, and nutritional parameters. This was a cross-sectional study of patients aged ≥18 years who had undergone dialysis for >3 months. Appetite was evaluated using the first 3 questions of the Appetite and Diet Assessment Tool (ADAT) questionnaire, which evaluate the appetite status during the past week as well as 1 question from the Kidney Disease and Quality of Life™ Short Form that assesses appetite in the past 4 weeks. The patients were divided into 3 groups according to the degree of appetite: group 1: very good and good appetite (ADAT) or not and somewhat (Kidney Disease and Quality of Life™ Short Form); group 2: fair or moderately; and group 3: poor and very poor, or very much and extremely. The nutritional parameters evaluated were body mass index (BMI), serum albumin, Geriatric Nutritional Risk Index (GNRI) and lean body mass index (lean mass in kilogram per square meter) as obtained by multifrequency bioelectrical impedance analysis. Patients with a BMI <23 kg/m(2), albumin <4 g/dL, GNRI <98, and lean body mass index below the lowest quartile were considered malnourished. The associations between appetite and nutritional variables were tested using Fisher exact test and by comparing the means of the variables in the 3 groups using the analysis of variance and Kruskal-Wallis tests. A total of 136 patients were included in the study with a mean age of 50.9 years and a median time on dialysis of 45 months; 57% of the patients were male. Regarding the first question on the ADAT questionnaire, 36% of patients exhibited anorexia in the past week. Furthermore, 28.7% of the population reported a lack of appetite in the last month. Moreover, 34.3% of the patients were considered malnourished according to BMI, 34.1% according to albumin, and 31.6% according to GNRI. Among the studied variables, the mean values of the following variables were different between groups 1, 2, and 3: hemoglobin (P = .0186), creatinine (P = .0392), albumin (P = .0065), GNRI (P = .0274), and lean BMI (P = .0274). The prevalence of a lack of appetite in hemodialysis patients in both the past week and the past month was high in the present study. The questionnaire evaluating appetite in the last 4 weeks could be used as a malnutrition screening tool in hemodialysis patients as suggested by the correlation of decreased appetite in the last month with variables that assess nutritional status such as albumin, lean body mass index, and GNRI.


Expression of human alpha 2-macroglobulin cDNA in baby hamster kidney fibroblasts: secretion of high levels of active alpha 2-macroglobulin.
Human alpha 2-macroglobulin (alpha 2M) is a unique 720-kDa proteinase inhibitor with a broad specificity. Unlike most other proteinase inhibitors, it does not inhibit proteolytic activity by blocking the active site of the proteinase. During complex formation with a proteinase, alpha 2M entraps the proteinase molecule in a reaction that involves large conformational changes in alpha 2M. We describe the molecular cloning of alpha 2M cDNA from the human hepatoblastoma cell line HepG2. The cDNA was subcloned under control of the adenovirus major late promoter in a mammalian expression vector and introduced into the baby hamster kidney (BHK) cell line. Transformed clones were isolated and tested for production of human alpha 2M with a specific enzyme-linked immunosorbent assay. Human recombinant alpha 2M (r alpha 2M), secreted and purified from isolated transfected BHK cell lines, was structurally and functionally compared to alpha 2M purified from human serum. The results show that r alpha 2M was secreted from the BHK cells as an active proteinase-binding tetramer with functional thiol esters. Cleavage reactions of r alpha 2M with methylamine and trypsin showed that the recombinant product, which was correctly processed at the N-terminus, exhibited molecular characteristics similar to those of the human serum derived reference. Moreover, r alpha 2M-trypsin complex bound to purified human placental alpha 2M receptor with an affinity indistinguishable from that of a complex formed from serum-derived alpha 2M and trypsin.


Mass Spectrometry-Sensitive Probes Coupled with Direct Analysis in Real Time for Simultaneous Sensing of Chemical and Biological Properties of Botanical Drugs.
The development of botanical materials as therapeutic agents involves the meticulous assessment of safety, efficacy, and quality. Compared with small-molecule drugs, quality control of botanical drugs confronts with more significant challenges due to their inherent complexity. Current quality control methods for botanical drugs, either prevailing chemical tests or emerging biological assays, are not able to meet recent demands of multiplexing, sensitivity, and speed. Here, we propose an on-demand strategy based on a direct analysis in real time-mass spectrometry (DART-MS) platform, which is capable of simultaneously analyzing multiple constituents and bioactivities of botanical drugs. Notably, the bioactivities are assessed by a multiple-enzyme assay that adopts cleavable mass spectrometry probes as enzymatic substrates: these probes labeled with a piperazine tag make possible sensitive, multiplexed, and quantitative enzyme activity measurements. The concept is successfully demonstrated via a case study of Danshen (Salvia miltiorrhiza) Injection where simultaneous detection of 34 constituents and inhibitory activities on two target enzymes can be achieved in just minutes. This proof-of-concept application also gives evidence that combining MS-sensitive probes with DART-MS can provide an environmentally friendly, highly sensitive analytical approach for botanical quality control.


Estimation of losses of iodine during different cooking procedures.
Iodine is an essential micronutrient. The human requirement of iodine is 150 mcg/ day. About 90% of this comes from food while 10% from the water. The most commonly used method of prophylaxis against iodine deficiency is via fortification of salt with iodine. At the beneficiary level iodised salt containing 15ppm of iodine is supplied. However, very few studies have been conducted to assess the losses of iodine during cooking procedures. Hence, a systematic study was undertaken with the objective to assess the losses of iodine during different cooking procedures. Fifty recipes commonly cooked in Indian families constituted the sample size. It was found that the mean losses of iodine during different procedures used was 1) pressure cooking 22%, 2) boiling 37%, 3) shallow frying 27%, 4) deep frying 20%, 5) roasting 6%, 6) steaming 20%. The findings of the present study indicate that further studies are needed in this field.


A Prediction Error-driven Retrieval Procedure for Destabilizing and Rewriting Maladaptive Reward Memories in Hazardous Drinkers.
Maladaptive reward memories (MRMs) can become unstable following retrieval under certain conditions, allowing their modification by subsequent new learning. However, robust (well-rehearsed) and chronologically old MRMs, such as those underlying substance use disorders, do not destabilize easily when retrieved. A key determinate of memory destabilization during retrieval is prediction error (PE). We describe a retrieval procedure for alcohol MRMs in hazardous drinkers that specifically aims to maximize the generation of PE and therefore the likelihood of MRM destabilization. The procedure requires explicitly generating the expectancy of alcohol consumption and then violating this expectancy (withholding alcohol) following the presentation of a brief set of prototypical alcohol cue images (retrieval + PE). Control procedures involve presenting the same cue images, but allow alcohol to be consumed, generating minimal PE (retrieval-no PE) or generate PE without retrieval of alcohol MRMs, by presenting orange juice cues (no retrieval + PE). Subsequently, we describe a multisensory disgust-based counterconditioning procedure to probe MRM destabilization by re-writing alcohol cue-reward associations prior to reconsolidation. This procedure pairs alcohol cues with images invoking pathogen disgust and an extremely bitter-tasting solution (denatonium benzoate), generating gustatory disgust. Following retrieval + PE, but not no retrieval + PE or retrieval-no PE, counterconditioning produces evidence of MRM rewriting as indexed by lasting reductions in alcohol cue valuation, attentional capture, and alcohol craving.


Monograph series on aging-related diseases: X. Prostate cancer.
Prostate cancer is the most commonly diagnosed cancer among Canadian men, excluding non-melanoma skin cancer. Prostate cancer incidence increases almost exponentially with age; most cases are diagnosed in men aged 65 years or older. With the possible exception of animal fat consumption, no known widespread modifiable risk factors have been identified. Although the prognosis is good if appropriate treatment occurs in the early stages of disease, the ability of existing early detection techniques to decrease mortality has not yet been demonstrated. The considerable economic and societal burden of prostate cancer and its treatment, coupled with the projected large increase in the number of new prostate cancer cases as the population ages, make this disease a very important public health issue.


Isomerization and oxygen atom transfer reactivity in oxo-Mo complexes of relevance to molybdoenzymes.
Both dioxo Mo(VI) and mono-oxo Mo(V) complexes of a sterically restrictive N2O heteroscorpionate ligand are found to exist as cis and trans isomers. The thermodynamically stable isomer differs for the two oxidation states, but in each case, we have isolated the kinetically labile isomer and followed its isomerization to the thermodynamically stable form. The Mo(VI) complex is more stable in the cis geometry and isomerizes more than 6 times faster than the Mo(V) complex, which prefers the trans geometry. In OAT reactions with PPh3, the trans isomer of the dioxo-Mo(VI) reacts approximately 20 times faster than the cis isomer. Thus, there are both oxidation state and donor atom dependent differences in isomeric stability and reactivity that could have significant functional implications for molybdoenzymes such as DMSO reductase.


Analyses of Genomic tRNA Reveal Presence of Novel tRNAs in Oryza sativa.
Transfer rRNAs are important molecules responsible for the translation event during protein synthesis. tRNAs are widespread found in unicellular to multi-cellular organisms. Analysis of tRNA gene family members in Oryza sativa revealed the presence of 750 tRNA genes distributed unevenly in different chromosomes. The length of O. sativa tRNAs genes were ranged from 66 to 91 nucleotides encoding 52 isoacceptor in total. tRNASer found in chromosome 8 of O. sativa encoded only 66 nucleotides which is the smallest tRNA of O. sativa and to our knowledge, this is the smallest gene of eukaryotic lineage reported so far. Analyses revealed the presence of several novel/pseudo tRNA genes in O. sativa which are reported for the first time. Multiple sequence alignment of tRNAs revealed the presence of family specific conserved consensus sequences. Functional study of these novel tRNA and family specific conserved consensus sequences will be crucial to decipher their importance in biological events. The rate of transition of O. sativa tRNA was found to be higher than the rate of transversion. Evolutionary study revealed, O. sativa tRNAs were evolved from the lineages of multiple common ancestors. Duplication and loss study of tRNAs genes revealed, majority of the O. sativa tRNA were duplicated and 17 of them were found to be undergone loss during the evolution. Orthology and paralogy study showed, the majority of O. sativa tRNA were paralogous and only a few of tRNASer were found to contain orthologous tRNAs.


Comparative evaluation of air cell and eggshell temperature measurement methodologies used in broiler hatching eggs during late incubation.
The current study was conducted to compare and contrast the uses of 2 devices (temperature transponder or infrared thermometer) and their locations (inner air cell membrane or outer eggshell surface) in Ross × Ross 708 broiler hatching eggs. The air cells of 14 embryonated and 10 nonembryonated eggs were implanted with temperature transponders on d 13.5 of incubation. Likewise, for these same eggs, eggshell surface temperature was detected with the use of transponders and an infrared thermometer. Temperatures were recorded every 12 h between 14.5 and 18 d of incubation, and graphs and corresponding regression values were used to track the temperatures over these time periods. The temperature readings using all methods in embryonated and nonembryonated eggs were positively correlated. In nonembryonated eggs, temperatures in the air cell and on the eggshell surface using transponders were higher than those on the eggshell surface using an infrared thermometer. Mean air cell temperature readings of embryonated eggs using transponders were higher than those of the eggshell, as determined with the use of transponders or an infrared thermometer. Furthermore, the differences in air cell temperature using transponders and eggshell temperature using an infrared thermometer in embryonated eggs increased with embryonic age. These readings confirmed increased embryo heat production during the incubational period examined. It was further concluded that when compared with actual embryo body temperatures determined in previous studies, the use of transponders in the air cells of broiler hatching eggs detected a higher and closer temperature than eggshell surface temperature. It is suggested that the air cell transponders in embryonated eggs circumvented the confounding effects of the thermal barrier properties of the eggshell and the flow of air across its surface.


Like Father, Like Son? Intergenerational Educational Mobility in India.
Using the nationally representative India Human Development Survey (IHDS), we create a unique son-father matched data set that is representative of the entire adult male population (aged 20-65) in India. We use these data to document the evolution of intergenerational transmission of educational attainment in India over time, among different castes and states for the birth cohorts of 1940-1985. We find that educational persistence, as measured by the regression coefficient of father's education as a predictor of son's education, has declined over time. This implies that increases in average educational attainment are driven primarily by increases among children of less-educated fathers. However, we do not find such a declining trend in the correlation between educational attainment of sons and fathers, which is another commonly used measure of persistence. To understand the source of such a discrepancy between the two measures of educational persistence, we decompose the intergenerational correlation and find that although persistence has declined at the lower end of the fathers' educational distribution, it has increased at the top end of that distribution.


Quadrupole collective modes in trapped finite-temperature Bose-Einstein condensates.
Finite-temperature simulations are used to study quadrupole excitations of a trapped Bose-Einstein condensate. We focus specifically on the m = 0 mode, where a long-standing theoretical problem is to account for an anomalous variation of the mode frequency with temperature. We explain this behavior in terms of the excitation of two separate modes, corresponding to the coupled motion of the condensate and the thermal cloud. The relative amplitude of the modes depends sensitively on the temperature and on the frequency of the harmonic drive used to excite them. Good agreement with experiment is found for appropriate drive frequencies.


Ischemic-reperfused rat skeletal muscle: the effect of vasoactive intestinal peptide (VIP) on contractile force, oxygenation and antioxidant enzyme systems.
The effect of vasoactive intestinal peptide (VIP) on the nerve-stimulated contraction, tissue oxygenation, lipid peroxidation and antioxidant enzymes activities-superoxide dismutase and catalase was investigated in the rat gastrocnemius muscle exposed to 4 h ischemia-4hr reperfusion. Ischemia caused significant decrease in muscle contractile force, oxygenation and superoxide dismutase enzyme activity. Reperfusion of ischemic muscle increased the muscle contractile force and restored the tissue oxygenation to the baseline level. Superoxide dismutase and catalase activities of reperfused muscle increased significantly. However neither ischemia nor reperfusion affected gastrocnemius muscle malondialdehide (MDA) levels. VIP administration at the onset of reperfusion significantly increased skeletal muscle contractile force and tissue oxygenation even higher than baseline and reperfusion values. VIP also normalized the increased superoxide dismutase and catalase activities of reperfused skeletal muscle. In conclusion, VIP, acting as a powerful antioxidant and preserving contractile machinery seems to be a promising endogenous peptide that can salvage the skeletal muscle from severe ischemia-reperfusion injury.


Assessment of the inhibition of Dengue virus infection by carrageenan via real-time monitoring of cellular oxygen consumption rates within a microfluidic device.
A microfluidic device combined with a light modulation system was developed to assess the inhibitory effect of carrageenan on Dengue virus (DENV) infection via real-time monitoring of cellular oxygen consumption rates (OCRs). Measuring cellular OCRs, which can reflect cellular metabolic activity, enabled us to monitor the process of viral infection in real time and to rapidly determine the antiviral activity of potential drugs/chemical compounds. The time variation of the cellular OCR of single cells that were infected in situ by DENV at different multiplicity of infection (m.o.i.) values was first successfully measured within a microfluidic device. The influence of the timing of carrageenan treatment on DENV infection was then examined by real-time monitoring of cellular OCRs in three groups. Cells that were pre-treated with carrageenan and then infected with DENV served as a pre-treatment group, cells to which carrageenan was added simultaneously with DENV served as a virucide group, and cells that were pre-infected with DENV and then treated with carrageenan served as a post-treatment group. By monitoring cellular OCRs, we could rapidly evaluate the inhibitory effect of carrageenan on DENV infection, obtaining a result within 7 h and showing that carrageenan had strong and effective anti-DENV activity in the three groups. In particular, a strong inhibitory effect was observed in the virucide group. Moreover, once the virus enters host cells in the post-treatment group, the immediate treatment with carrageenan for the infected cells has higher efficiency of antiviral activity. Our proposed platform enables to perform time-course or dose-response measurements of changes in cellular metabolic activity caused by diseases, chemical compounds, and drugs via monitoring of the cellular OCR, with rapid and real-time detection. This approach provides the potential to study a wide range of biological applications in cell-based biosensing, toxicology, and drug discovery.


Bioprospecting Through Cloning of Whole Natural Product Biosynthetic Gene Clusters.
Since the discovery of penicillin, natural products and their derivatives have been a valuable resource for drug discovery. With recent development of genome mining approaches in the post-genome era, a great number of natural product biosynthetic gene clusters (BGCs) have been identified and these can potentially be exploited for the discovery of novel natural products that can find application as pharmaceuticals. Since many BGCs are silent or do not express in native hosts under laboratory conditions, heterologous expression of BGCs in genetically tractable hosts becomes an attractive route to activate these BGCs to discover the corresponding products. Here, we highlight recent achievements in cloning and discovery of natural product biosynthetic pathways via intact BGC capturing, and discuss the prospects of high-throughput and multiplexed cloning of rational-designed gene clusters in the future.


Impact of perinatal dioxin exposure on infant growth: a cross-sectional and longitudinal studies in dioxin-contaminated areas in Vietnam.
Dioxin exposure levels remain elevated in residents living around former US Air Force bases in Vietnam, indicating potential adverse impacts on infant growth. In this study, 210 mother-infant pairs in dioxin-contaminated areas in Vietnam were recruited at the infants' birth and followed up for 4 months. Perinatal dioxin exposure levels were estimated by measurement of polychlorinated dibenzo-p-dioxins/furans toxic equivalent (PCDDs/Fs-TEQ) in breast milk. The infants' size was measured at birth and 1 and 4 months after birth, and neurodevelopment was evaluated using the Bayley Scales III at 4 months of age. Among 4 dioxin groups (<25, 25-50, 50-75, ≥75 percentile of PCDDs/Fs-TEQ), cross-sectional comparisons of body size and neurodevelopment scales and comparisons of longitudinally assessed body size were performed respectively. At birth, head circumference of girls in the ≥75 percentile group was significantly larger than those in the <25 and 50-75 percentile groups. At 4 months of age, the weight and body mass index (BMI) of boys in the ≥75 percentile group were significantly lower than those in the other groups. Increase in weight was significantly lower in the ≥75 percentile group in both sexes from birth to 1 month but only in boys at 1-4 months of age. Estimated marginal mean values in a mixed model of weight and BMI during the first 4 months of life were significantly lower in the ≥75 percentile group in boys. In girls, marginal mean values for head circumference were increased with increase in dioxin levels. Only in boys, cognitive, language, and fine motor scores in the ≥75 percentile group were significantly lower than those in the other groups. These results suggested a considerable impact of perinatal dioxin exposure on infant growth, particularly in boys exposed to dioxins at high level of PCDDs/Fs-TEQ.


A model-based approach for detecting and identifying faults on the D.C. side of a P.V. system using electrical signatures from I-V characteristics.
With the development of distributed generation and the corresponding importance of the P.V. (photovoltaic) system, it is desired to operate a P.V. system efficiently and reliably. To ensure such an operation, a monitoring system is required to diagnose the health of the system. This paper aims to analyze a P.V. system under various operating conditions to identify parameters-derived from the I-V (current-voltage) characteristics of the P.V. system-that could serve as electrical signatures to various faulty operations and facilitate in devising a monitoring algorithm for the system. A model-based approach has been adopted to represent a P.V. system, using a one-diode model of a practical P.V. cell, developed in MATLAB/Simulink. The modelled system comprises two arrays, while each array has two panels in series. It was simulated for various operating conditions: healthy condition represented by STC (Standard Testing Condition), O.C. (open-circuited), soiling, P.S. (partial-shading), H.S. (panels hotspots) and P.D. (panels degradation) conditions. For the analysis of I-V curves under these conditions, six derived parameters were selected: Vte (equivalent thermal voltage), MCPF (maximum current point factor), Ri (currents ratio), S (slope), and Dv and Di (voltages and currents differences, respectively). Using these parameters, data of the actual system under various conditions were compared with its model-generated data for healthy operating conditions. Thresholds were set for each parameter's value to mark normal operation range. It was observed that almost each considered fault creates a unique combination of sensitive parameters whose values exceeds the pre-defined thresholds, creating an electrical signature that will appear only when the corresponding conditions on the system are achieved. Based on these signatures, an algorithm has been proposed in this study which aims to identify and classify the considered faults. In comparison to other such studies, this work has been focused on those sensitive parameters for faults identification which shows greater sensitivity and contribute more to creation of unique sets of sensitive parameters for considered faults.


Mechanical stimulation of energetic materials at the nanoscale.
The initiation of energetic materials by mechanical stimuli is a critical stage of their functioning, but remains poorly understood. Using atomic force microscopy (AFM) we explore the microscopic initiation behavior of four prototypical energetic materials: 3,4-dinitropyrazole, ε-CL-20, α-PETN and picric acid. Along with the various chemical structures, these energetic compounds cover a range of application types: a promising melt-cast explosive, the most powerful energetic compound in use, a widespread primary explosive, and a well-established nitroaromatic explosive from the early development of energetics. For the softest materials (picric acid and 3,4-dinitropyrazole), the surfaces were found to behave dynamically, quickly rearranging in response to mechanical deformation. The pit created by nanoscale friction stimulation on the surface of 3,4-dinitropyrazole doubled in volume upon aging for half an hour. Over the same time frame, a similar pit on the picric acid surface increased in volume by more than seven-fold. Remarkably, increased humidity was found to reduce the rate of surface rearrangement, potentially offering an origin for the desensitization of energetic materials when wetted. Finally, we identify an inverse correlation between the surface dynamics and mechanical sensitivity of our test energetic compounds. This strongly suggests that surface dynamics influence a material's ability to dissipate excess energy, acting as a buffer towards mechanical initiation.


Thyrotropin receptor antibody activities significantly correlate with the outcome of radioiodine (131I) therapy for hyperthyroid Graves' disease.
The outcome of 131I therapy for 109 patients with Graves' disease was analysed according to pretreatment laboratory data including thyrotropin receptor antibody (TRAb) activities. Forty-five percent of patients became euthyroid, and 13% of patients became hypothyroid within one year after 131I therapy. Forty-two percent of patients remained hyperthyroid one year after 131I therapy. Pretreatment values for serum T4, T3, and the estimated weight of the thyroid were significantly higher in the hyperthyroid group. The mean for the TRAb index of the hyperthyroid group was significantly higher than that of the euthyroid group. Life table analysis revealed a significant effect of the TRAb index on the rate of hyperthyroidism after 3 months or later. These results appear to suggest that the TRAb index is one of the factors which influence the outcome of 131I therapy for Graves' disease.


[Clinical experience with cefoxitin in the surgical field].
Cefoxitin (CFX) was administered to 49 hospitalized patients in the surgical field, and the following results were obtained: Clinical results of the 12 patients with surgical infections were excellent in 1 patient, good in 10, and poor in 1, with the efficacy rate of 91.7%. CFX was also administered to 37 patients for prophylaxis of postoperative infections, and the clinical efficacy rate was 91.7%. No side effects were seen besides mild eruption in 1 patient. The above results indicate that CFX is exceeding useful in surgical field.


Comparison of phenotypically indistinguishable but geographically distinct Neisseria meningitidis Group B isolates in a serum bactericidal antibody assay.
The "gold standard" assay for measuring serologic protection against Neisseria meningitidis group B (MenB) is the serum bactericidal antibody (SBA) assay. Of vital importance to the outcome of the SBA assay is the choice of the target strain(s), which is often chosen on the basis of phenotype or genotype. We therefore investigated the effect on the results produced by the SBA assay of using phenotypically indistinguishable but geographically distinct MenB isolates. Nine PorA P1.19,15 and 11 PorA P1.7-2,4 MenB isolates were incorporated into the SBA assay using human complement and were assayed against sera obtained either before or after outer membrane vesicle vaccination. Large differences in the results produced by the isolates in the SBA assay were demonstrated. These included differences as great as 5.8-fold in SBA geometric mean titers and in the proportions of subjects with SBA titers of >/=4. Ranges of as many as 9 SBA titers were achieved by individual sera across the panels of isolates. To determine the reasons for the differences observed, investigations into the expression of capsular polysaccharide, PorA, PorB, Opc, and lipooligosaccharide (LOS) and into LOS sialylation were completed. However, minor differences were found between strains, indicating similar expression and no antigen masking. These results have implications for the choice of MenB target strains for inclusion in future studies of MenB vaccines and highlight the requirement for standardization of target strains between laboratories.


Forced Air Warming System: Evaluation of Internal System Contamination.
Despite the indisputable need to prevent peri-operative hypothermia, some evidence in the literature questions the possible role of forced air warming systems as a risk factor for the occurrence of surgical site infection. The objective of the study is to evaluate the micro-biologic safety of a forced air warming systems (FAW) in relation to the risk of emission of micro-organisms in the surgical environment. A quantitative, descriptive-exploratory laboratory study performed in a large hospital. An evaluation of possible internal contamination was performed by collecting air from the hose onto plates containing Trypticase Soy Agar. The experiment was performed in triplicate two months after replacing the filter and again after six months using a random sample of 50% of the FAW in use, which corresponded to 13 FAW and a total of 75 samples. Among the 39 samples analyzed in the first stage, only nine (23.1%) plates presented microbial growth of one or two colonies, while only six (16.7%) plates of the 36 evaluated samples in the second phase had growth of one or two colonies. This study showed small microbial growth of culture after 48 hours after filter replacement. New investigations that correlate the findings of micro-biology analysis and the occurrence of surgical site infection should be conducted.


Oleic acid stimulates HC11 mammary epithelial cells proliferation and mammary gland development in peripubertal mice through activation of CD36-Ca2+ and PI3K/Akt signaling pathway.
This study aimed to investigate the effects of oleic acid (OA), a monounsaturated fatty acid, on HC11 mammary epithelial cells proliferation and peripubertal mammary gland development and explore the underlying mechanisms. HC11 cells and C57BL/6J mice were treated with OA. HC11 proliferation, peripubertal mammary gland development, and the involvement of CD36 and PI3K/Akt were assessed. In vitro, 100 μM OA significantly promoted HC11 proliferation by increasing Cyclin D1/3 and PCNA expression and decreasing p21 expression. Meanwhile, OA enhanced CD36 expression, elevated [Ca2+]i and activated PI3K/Akt signaling pathway. However, knockdown of CD36, chelation of [Ca2+]i or inhibition of PI3K eliminated the OA-induced promotion of HC11 proliferation and change in proliferative markers expression. In vivo, peripubertal exposure to diet containing 2% OA stimulated mammary duct development, with increased terminal duct end (TDE) and ductal branch. Moreover, dietary OA increased the serum levels of IGF-1 and E2, enhanced the expression of CD36 and Cyclin D1, and activated PI3K/Akt pathway in mammary glands. In conclusion, OA stimulated HC11 cells proliferation and mammary gland development in peripubertal mice, which was associated with activation of CD36-[Ca2+]i and PI3K/Akt signaling pathway. These data provided new insights into the stimulation of mammary gland development by dietary oleic acid.


Effects of novel phenoxazine compounds, 2-amino-4, 4a-dihydro-4alpha, 7-dimethyl-3H-phenoxazine-3-one and 3-amino-1, 4alpha-dihydro-4alpha, 8-dimethyl-2H-phenoxazine-2-one on proliferation of phytohemagglutinin- or anti-human IgM-activated human peripheral blood mononuclear cells.
We examined the in vitro effects of 2-amino-4, 4alpha-dihydro-4alpha, 7-dimethyl-3H-phenoxazine-3-one(Phx 1)and 3-amino-1, 4alpha-dihydro-4a, 8-dimethyl-2H-phenoxazine-2-one (Phx 2) on the proliferation of phytohemagglutinin (PHA)- or anti-human IgM-activated human peripheral blood mononuclear cells (PBMC). Phx 1 and Phx 2 inhibited the incorporation of 3H-thymidine of PHA-activated PBMC by as much as 75% and 40%, respectively, at a concentration of 40 microM. The inhibition was dependent on the dose of Phx 1 and Phx 2. These results strongly suggest that Phx 1 and Phx 2 inhibit proliferation of T cells, because PHA specifically activates the T cells among PBMC. On the other hand, when PBMC were activated by anti-human IgM, which specifically stimulates B cells, the incorporation of 3H-thymidine was rather increased in the presence of 15.8 microM Phx 1 or Phx 2. However, at a higher concentration of Phx 1 or Phx 2 (50 microM), the incorporation of 3H-thymidine was increased by Phx 1, but was inhibited by Phx 2. These results suggest different effects of Phx 1 and Phx 2 on proliferation of human T and B cells.


Regularized orbital-optimized second-order perturbation theory.
Orbital-optimized second-order perturbation theory (OOMP2) optimizes the zeroth order wave function in the presence of correlations, removing the dependence of the method on Hartree-Fock orbitals. This is particularly important for systems where mean field orbitals spin contaminate to artificially lower the zeroth order energy such as open shell molecules, highly conjugated systems, and organometallic compounds. Unfortunately, the promise of OOMP2 is hampered by the possibility of solutions being drawn into divergences, which can occur during the optimization procedure if HOMO and LUMO energies approach degeneracy. In this work, we regularize these divergences through the simple addition of a level shift parameter to the denominator of the MP2 amplitudes. We find that a large level shift parameter of 400 mEh removes divergent behavior while also improving the overall accuracy of the method for atomization energies, barrier heights, intermolecular interactions, radical stabilization energies, and metal binding energies.


Psychological Impact of COVID-19 Patient Care on Families of Healthcare Workers.
Objective: The Coronavirus‑19 (COVID‑19) pandemic has perpetrated a significant mental health burden amongst health care workers (HCWs) and their families. Families of HCWs are at significant risk of various mental health problems due to concerns regarding the wellbeing of the HCW, fear of contacting the virus from HCWs, staying isolated during periods of quarantine and being the passive recipient of public stigma directed towards HCWs. The objective of this study was to assess the perceived stress, resilience and coping tendencies of families of HCWs managing the COVID 19 crisis. Method : A cross sectional study was conducted at a tertiary care hospital in India, where close relatives of 150 HCWs managing COVID-19 patients (directly or indirectly involved in patient care) were selected using systematic random sampling. They were evaluated using the Perceived Stress Scale, Brief Resilience Coping Scale, and the Hospital Anxiety and Depression Scale. Pearson's correlation and Independent t-test were used for statistical analysis using the SPSS software. Results: Majority (75%) of the family members associated with frontline HCWs had moderate to high level of perceived stress, while23% and 17% had clinically significant anxiety and depressive symptoms, respectively. More than 50% had low resilience and coping scores. High levels of stress, anxiety and depression were seen in female respondents, those less than 40 years old, having a child or an elderly family member and when the HCW had direct involvement in COVID-19 patient care. Conclusion: Families of HCWs suffer from significant psychological burden due to the COVID-19 crisis. They face significant risk of depression and anxiety associated problems, which are augmented by low resilience and impaired coping. Mental health of HCWs and their family members should be given due attention with a focus on early identification, providing psychological support and improving resilience.


In Silico Study, Synthesis, and Cytotoxic Activities of Porphyrin Derivatives.
Five known porphyrins, 5,10,15,20-tetrakis(p-tolyl)porphyrin (TTP), 5,10,15,20-tetrakis(p-bromophenyl)porphyrin (TBrPP), 5,10,15,20-tetrakis(p-aminophenyl)porphyrin (TAPP), 5,10,15-tris(tolyl)-20-mono(p-nitrophenyl)porphyrin (TrTMNP), 5,10,15-tris(tolyl)-20-mono(p-aminophenyl)porphyrin (TrTMAP), and three novel porphyrin derivatives, 5,15-di-[bis(3,4-ethylcarboxymethylenoxy)phenyl]-10,20-di(p-tolyl)porphyrin (DBECPDTP), 5,10-di-[bis(3,4-ethylcarboxymethylenoxy)phenyl]-15,20-di-(methylpyrazole-4-yl)porphyrin (cDBECPDPzP), 5,15-di-[bis(3,4-ethylcarboxymethylenoxy)phenyl]-10,20-di-(methylpyrazole-4-yl)porphyrin (DBECPDPzP), were used to study their interaction with protein targets (in silico study), and were synthesized. Their cytotoxic activities against cancer cell lines were tested using 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolium bromide (MTT) assay. The interaction of porphyrin derivatives with carbonic anhydrase IX (CAIX) and REV-ERBβ proteins were studied by molecular docking and molecular dynamic simulation. In silico study results reveal that DBECPDPzP and TrTMNP showed the highest binding interaction with REV- ERBβ and CAIX, respectively, and both complexes of DBECPDPzP-REV-ERBβ and TrTMNP-CAIX showed good and comparable stability during molecular dynamic simulation. The studied porphyrins have selective growth inhibition activities against tested cancer cells and are categorized as marginally active compounds based on their IC50.


Effectiveness of coral relocation as a mitigation strategy in Kāne'ohe Bay, Hawai'i.
Coral reef restoration and management techniques are in ever-increasing demand due to the global decline of coral reefs in the last several decades. Coral relocation has been established as an appropriate restoration technique in select cases, particularly where corals are scheduled for destruction. However, continued long-term monitoring of recovery of transplanted corals is seldom sustained. Removal of coral from a navigation channel and relocation to a similar nearby dredged site occurred in 2005. Coral recovery at the donor site and changes in fish populations at the receiving site were tracked periodically over the following decade. Coral regrowth at the donor site was rapid until a recent bleaching event reduced coral cover by more than half. The transplant of mature colonies increased spatial complexity at the receiving site, immediately increasing fish biomass, abundance, and species that was maintained throughout subsequent surveys. Our research indicates that unlike the majority of historical accounts of coral relocation in the Pacific, corals transplanted into wave-protected areas with similar conditions as the original site can have high survival rates. Data on long-term monitoring of coral transplants in diverse environments is central in developing management and mitigation strategies.


Molecular identification of the potentially forensically relevant cluster flies Pollenia rudis (Fabricius) and Pollenia vagabunda (Meigen) (Diptera: Polleniidae) - non-recorded species in Algeria.
Cluster flies are represented by the genus Pollenia Robineau-Desvoidy, 1830 of the family Polleniidae Brauer and Bergenstamm, 1889. Their larvae are known to be internal parasites or predators of earthworms. Herein, we report for the first time the occurrence of the cluster flies Pollenia rudis Fabricius, 1794 and Pollenia vagabunda (Meigen, 1826) (Diptera: Polleniidae) on carcasses in Algeria and identify them through DNA barcoding. A region of the mitochondrial cytochrome c oxidase I gene (COI) was amplified and sequenced. Genetic distances were determined. A phylogenetic tree was constructed with the maximum parsimony method using 10 000 bootstrap replicates. A total number of 157 adults of P. rudis were collected together with 325 adults of Pollenia vagabunda. The occurrence of Pollenia on animal carcasses does not seem to be correlated with a particular stage of decomposition. All the sequences were correctly identified using the BLASTn tool from the GenBank database and the BOLD identification engine. Intra- and interspecific sequence divergence values were less than 1% and greater than 3%, respectively. COI barcodes obtained from this study were robust enough to identify and distinguish unambiguously between P. rudis and P. vagabunda. In the tree-based analysis, the cluster flies were all assigned to their respective species separately from each other confirming the morphological identification. These results provide DNA barcodes that contribute to the growth of reference databases and allow fast and accurate identification.


Crystallization of a genetically engineered water-soluble primary penicillin target enzyme. The high molecular mass PBP2x of Streptococcus pneumoniae.
A genetically engineered water-soluble derivative of PBP2x of Streptococcus pneumoniae has been produced, purified and crystallized in a form suitable for X-ray diffraction analysis. The best crystals have been grown at 15 degrees C, from solutions containing 8% polyethylene glycol 10,000 at pH values ranging from 3.9 to 6.0. These crystals diffract to a resolution of 3.5 A and have a space group P6(1)22 (or enantiomorph) with unit cell dimensions of a = b = 162.2 A, c = 171.8 A, alpha = beta = 90 degrees, gamma = 120 degrees. The molecular mass and cell dimensions suggest that there is one molecule of enzyme per asymmetric unit. The breakdown of a chromogenic cephalosporin derivative diffused into a crystal reveals clearly that the enzyme is active in the crystalline state.


The impact of one session of HF-rTMS on salivary cortisol in healthy female subjects.
Previous studies in healthy volunteers reported a possible impact of high frequency repetitive transcranial magnetic stimulation (HF-rTMS) on stress hormones, like cortisol. In this sham-controlled, "single blind", crossover study, we examined whether HF-rTMS had an effect on the hypothalamic-pituitary-adrenal (HPA) axis, by analysing salivary cortisol levels. Two studies were conducted. First, HF-rTMS on the left dorsolateral prefrontal cortex (DLPFC) was performed in 28 young healthy female volunteers. Second, in a comparable, but different group of 26 healthy females, HF-rTMS was performed on the right DLPFC. Salivary cortisol levels were assessed before, immediately after and 30 min after real and sham HF-rTMS. We found no support for the hypothesis that one single session of HF-rTMS on the left or the right DLPFC has an immediate or delayed impact on the HPA-axis, as measured by salivary cortisol. Although we controlled for several methodological problems in HF-rTMS research, the hypothesis that one single session of HF-rTMS on the left or on the right DLPFC can influence the HPA-axis in healthy volunteers was not supported.


[Action of hyperoxia on the total electrical activity of the olfactory epithelium of the frog].
Olfactory mucosa of the frog was stimulated with odours of amylacetat , butanol, alcohol, naphthalene, acetic acid in air and oxygen media under normal and high oxygen pressure (0.2 MPa and 0.7 MPa). The electroolfactogram (EOG) amplitude was higher in oxygen due, probably, to the olfactory mucosa oxygen saturation on concentration gradient as well as to the activation of processes related to the EOG generation. 0.2 MPa pressure as well as 0.7 MPa pressure applied for 60 min decreased the EOG amplitude.


[Co-existence and co-release of gastrin 34 N-terminal fragment with gastrin 17 in rat stomach].
Big gastrin comprising 34 amino acid residues (G34) consists of an N-terminal pentadecapeptide linked via two lysine residues to a C-terminal heptadecapeptide identical with little gastrin (G17). Both G17 and G34 have now been established as the principal active forms of gastrin. In this study, release of G34 N-terminal peptide fragment of methacholine and porcine gastrin releasing peptide (pGRP) stimulation in isolated rat stomach perfusion system was investigated by radioimmunoassay with use of an antiserum specific to the N-terminal portion of G34. G34 N-terminal immunoreactivity (IR-G34-N) was detected in rat stomach and proximal duodenum, and the highest concentration was found in extract of the antral mucosa. The concentration of IR-G34-N was constantly lower than that of IR-G17. By gel-filtration study, IR-G34-N in antral mucosa extract was attributed mostly to the G34 N-terminal pentadecapeptide-like component, and the concentration of G34 was about one tenth of G17. Methacholine 10(-8)-10(-3) M produced a biphasic dose-dependent release of IR-G34-N from the vascularly perfused isolated rat stomach. The maximal release was shown by 10(-5) M of methacholine. The release was concomitant with that of IR-G17 during methacholine stimulation. Stimulation of pGRP (14-27) (10(-7) M) produced a monophasic release of IR-G34-N from the vascularly perfused isolated rat stomach. The release was concomitant with that of G17 during the stimulation. The integrated IR-G34-N release was not stoichiometric with that of IR-G17, and IR-G34-N was constantly low. Gel-filtration of the perfusate from rat stomach revealed the presence of the G34 N-terminal pentadecapeptide-like component as a sole major component. The present results demonstrate that post-translational processing of the gastrin precursor in the rat antrum did not necessarily produce G34, which is further converted in the tissue to G17-related peptide(s) and that the G34 N-terminal fragment formed in the G34 conversion is stored and released concomitantly with G17-related peptide(s).


Communication about end-of-life care between language-discordant patients and clinicians: insights from medical interpreters.
Communication about health care and especially end-of-life care is difficult for clinicians and patients when they do not speak the same language. Our purpose was to improve understanding of how to approach discussions between language-discordant patients and clinicians about terminal or life-threatening illness. We conducted a qualitative study with 4 focus groups with 43 professional medical interpreters. We asked open-ended questions concerning physician and interpreter communication about end-of-life care. Focus groups were audiotaped, transcribed, and analyzed using principles of grounded theory. Results were presented to an additional 3 focus groups with 25 medical interpreters to ensure that analyses represented interpreters' perspectives. We developed 3 frameworks for understanding high-quality language-discordant communication about end-of-life care. The first framework addresses physician and interpreter professionalism, including humanistic qualities and emotional support capabilities important for high quality care. The second framework is physician-centered and highlights communication skills, as well as coordination with other providers and cultural sensitivity. The third framework is interpreter-centered, focusing on role conflicts, including struggles concerning expectations to provide strict interpretation versus being a cultural broker. Interpreters' recommendations for improving quality of this care include pre-meetings with interpreters before encounters involving delivery of bad news and explicit discussions with interpreters about whether the clinician expects strict interpretation or cultural brokering. These results provide insights for physicians about how to improve end-of-life discussions with language-discordant patients and their families. Interpreter recommendations provide physicians and health care organizations with specific tools that may improve quality of communication about end-of-life discussions.


Chronic hepatitis C in childhood: an 18-year experience.
The long-term outcome of chronic hepatitis C (CHC) has not been well studied, both for untreated and interferon-treated children. The aim of this study was to evaluate the long-term outcome of disease in a large series of children with CHC. Clinical, biochemical, virological, and histological features were evaluated in all children (age, 2-18 years) with CHC who did not have concomitant disease and who attended at our hospital's liver unit during the period of 1986-2004. One hundred twenty-five children with CHC were studied. All patients remained free of symptoms throughout the period of observation. On the basis of transaminase levels during the first year of positivity for antibodies to hepatitis C virus (HCV), children were divided into 2 groups: patients with hypertransaminasemia (100 patients, all of whom had detectable HCV RNA), and those with normal transaminases (25 patients; 16 had viremia and 9 did not have viremia). Sustained clearance of viremia was achieved in 38% of the patients treated with interferon, compared with 12% of untreated children (P<.05). A sustained response to therapy was obtained in 64.7% of children infected with an HCV genotype other than genotype 1 and in 24.2% of those infected with HCV genotype 1 (P<.05). Histological lesions were mild in all 64 patients who underwent liver biopsy. No linear correlation was found between duration of disease and progression of fibrosis. Examination of a follow-up liver biopsy specimen revealed cirrhosis only in 1 (4.7%) of 21 children. Children with CHC were symptom free and had a morphologically mild liver disease. Interferon therapy may be effective for patients infected with HCV genotypes other than genotype 1, whereas lower response rates are expected for HCV genotype 1-infected children. The real impact of therapy on long-term outcome remains to be established.


Optimization of batch processes involving simultaneous enzymatic and microbial reactions.
Two general models for batch simultaneous enzymatic and microbial reaction (SEMR) processes are presented, the second derived from and simpler than the first and accounting for enzyme denaturation. Using the second model and parameter values from the literature, simulation was used to examine a range of enzyme addition rate strategies (in which the rate was a linear function of time) for a relatively fast ethanol fermentation and for a longer duration citric acid fermentation, both using cellulose as the substrate. For the ethanol process it is optimal (for a specific objective function which accounts for product value and enzyme cost) to add all the enzyme at the beginning of the process. But for the citric acid process a linearly decreasing enzyme addition rate, coupled with the addition of a small fraction of the enzyme at time zero, is better than pure batch operation or operation with the best constant enzyme feed rate.