The isolation of natural killer (NK)-resistant variants of the K562 cell line by mutagenesis and selection with antibodies which inhibit NK cell-mediated lysis.
Mechanisms involved in the lysis of tumor cells by natural killer (NK) cells were investigated by using mutagenized K562 targets resistant to the effects of NK cells. K562 cells were treated with the mutagen methyl methanesulfonate (MMS) and, to select for resistant mutants, rabbit anti-idiotypic (anti-id) antibodies were used. This anti-id was raised to a monoclonal antibody 9.1C3 which itself blocked lysis by NK cells by binding to the effector cells; the anti-id inhibited killing by binding to the K562 targets, presumably to a cell surface protein relevant to a secondary event in the NK lytic pathway. MMS-derived mutants showed a heterogeneity of staining with the anti-id, allowing the antibody to be used with flow cytometry to select a population of K562 cells relatively negative in antigen expression. The degree of reactivity of K562 cultures with the anti-id antiserum and the resistance to lysis by NK cells were inversely related. Cultures of NK-resistant K562 cells with low expression of the anti-id structure were cloned by limiting dilution: 96 clones were analyzed and one subclone, C9/2, which was six-to sevenfold less sensitive to lysis than the parental K562 cell line, was used in further studies by cold target inhibition and single cell binding assays. The increased resistance to lysis of C9/2 was not due to a reduced expression of target recognition structures, and resistance could not be overcome by prolonging the time allowed for lysis to 18 hr nor by adding exogenous recombinant leukocyte interferon. Killing of the NK-resistant variant was inhibited by mannose-6-phosphate but not by the monoclonal antibody against which the anti-id antibody was raised. It is therefore suggested that the structure on the K562 cells recognized by the anti-id antibodies is a novel secondary receptor which is important in the later stages of the NK cell cytolytic cascade.


Quantitative freeze-fracture and filipin-binding study of retinal pigment epithelial-cell basal membranes in diabetic rats.
Breakdown of the blood-retinal barrier in diabetes may be related to alterations in the retinal pigment epithelial (RPE) cell layer. Morphological studies suggest increased permeability of diabetic RPE plasma membranes, and proliferation and flattening of the RPE basal infoldings have been observed in diabetic animals. In order to determine whether these phenomena are associated with changes in membrane protein or sterol composition, we have used quantitative electron-microscope freeze-fracture and filipin-binding techniques to study the RPE basal membrane in streptozotocin diabetic and 3-O-methyl glucose control rats. Perfusion-fixed retinas were processed for freeze-fracture and filipin-binding analysis. Filipin, a polyene antibiotic, binds specifically to 3-beta-hydroxy-sterols to produce membrane deformations recognizable by freeze-fracture. These analyses revealed an 11% increase in the density of intramembrane particles within the cytoplasmic (P-face) leaflet in diabetic rats as compared with the controls (P less than 0.01, t test). The increase occurred primarily in 6-9-nm particles, while smaller particles were decreased (P less than 0.001, chi-square test). Filipin binding was the same in both groups. These results suggest that alterations in intrinsic membrane proteins may contribute to permeability and surface area changes in the diabetic RPE but that RPE membrane sterols are not affected by diabetes.


Endovascular treatment with in-vitro fenestration and sac filling technique for ruptured thoracoabdominal aortic aneurysm with Behcet's disease.
We provided an endovascular strategy of treating ruptured aortic aneurysm with Behcet's Disease. A 25-year-old man was diagnosed ruptured thoracoabdominal aortic aneurysm with Behcet's Disease according to his eye damage history, high level of ESR and C-reactive protein and the imaging result. We used in-vitro fenestration of the stent-graft combined with in-stent technique to occlude the ruptured aortic aneurysm and preserve the blood supply from the aorta for visceral arteries in emergency. Sac filling technique was used to treat the endoleak to quickly prevent bleeding. The patient kept post-operative immunotherapy for 1 year. The patient had a good prognosis in the reduction of the cavity of aortic aneurysm to the normal size without any complications in a year follow-up.


Influenza Vaccine Effectiveness in the United States During the 2016-2017 Season.
In recent influenza seasons, the effectiveness of inactivated influenza vaccines against circulating A(H3N2) virus has been lower than against A(H1N1)pdm09 and B viruses, even when circulating viruses remained antigenically similar to vaccine components. During the 2016-2017 influenza season, vaccine effectiveness (VE) across age groups and vaccine types was examined among outpatients with acute respiratory illness at 5 US sites using a test-negative design that compared the odds of vaccination among reverse transcription polymerase chain reaction-confirmed influenza positives and negatives. Among 7083 enrollees, 1342 (19%) tested positive for influenza A(H3N2), 648 (9%) were positive for influenza B (including B/Yamagata, n = 577), and 5040 (71%) were influenza negative. Vaccine effectiveness was 40% (95% confidence interval [CI], 32% to 46%) against any influenza virus, 33% (95% CI, 23% to 41%) against influenza A(H3N2) viruses, and 53% (95% CI, 43% to 61%) against influenza B viruses. The 2016-2017 influenza vaccines provided moderate protection against any influenza among outpatients but were less protective against influenza A(H3N2) viruses than B viruses. Approaches to improving effectiveness against A(H3N2) viruses are needed.


Separable substrates for anticipatory and consummatory food chemosensation.
Perception of the smell of a food precedes its ingestion and perception of its flavor. The neurobiological underpinnings of this association are not well understood. Of central interest is whether the same neural circuits code for anticipatory and consummatory phases. Here, we show that the amygdala and mediodorsal thalamus respond preferentially to food odors that predict immediate arrival of their associated drink (FO+) compared to food odors that predict delivery of a tasteless solution (FO-) and compared to the receipt of the drink. In contrast, the left insula/operculum responds preferentially to the drink, whereas the right insula/operculum and left orbitofrontal cortex respond to FO+ and drink. These findings indicate separable and overlapping representation of anticipatory and consummatory chemosensation. Moreover, since ratings of perceived pleasantness of FO+, FO-, and drink were similar, the response in the amygdala and thalamus cannot reflect acquired affective value but rather predictive meaning or biological relevance.


Development and testing of a new framework for rapidly assessing legal and managerial protection afforded by marine protected areas: Mediterranean Sea case study.
The Convention on Biological Diversity (CBD) states the need to effectively conserve at least 10% of coastal and marine areas of particular importance for biodiversity by 2020. Here, a new indicator-based methodological framework to assess biodiversity protection afforded by marine protected areas' (MPA) was developed as a quick surrogate for MPAs' potential conservation effectiveness: the Marine Protected Area Protection Assessment Framework (MaPAF). The MaPAF consists of a limited number of headline indicators that are integrated in two indexes: Legal protection and Management effort, which eventually integrate in the overall MPA Protection super-index. The MaPAF was then tested in the Mediterranean MPA network as a case study. Spatial analyses were performed at three meaningful scales: the whole Mediterranean Sea, Mediterranean ecoregions and countries. The results of this study suggest that: 1) The MaPAF can serve as a useful tool for consistent, adaptive, quick and cost-effective MPA effectiveness assessments of MPAs and MPA networks in virtually any marine region, as the headline indicators used are commonly compiled and easy to retrieve; 2) The MaPAF proved usable and potentially relevant in the Mediterranean Sea where most indicators in the framework can be publicly accessed through the MAPAMED database and are planned to be regularly updated; 3) Protection afforded by MPAs is low across the whole Mediterranean, with only few MPAs having relatively high legal and managerial protection; and 4) Most Mediterranean countries need to devote substantially more work to improve MPA effectiveness mainly through increased management effort.


Signal transfer in haloarchaeal sensory rhodopsin- transducer complexes.
Membrane-inserted complexes consisting of two photochemically reactive sensory rhodopsin (SR) subunits flanking a homodimer of a transducing protein subunit (Htr) are used by halophilic archaea for sensing light gradients to modulate their swimming behavior (phototaxis). The SR-Htr complexes extend into the cytoplasm where the Htr subunits bind a his-kinase that controls a phosphorylation system that regulates the flagellar motors. This review focuses on current progress primarily on the mechanism of signal relay within the SRII-HtrII complexes from Natronomonas pharaonis and Halobacterium salinarum. The recent elucidation of a photoactive site steric trigger crucial for signal relay, advances in understanding the role of proton transfer from the chromophore to the protein in SRII activation, and the localization of signal relay to the membrane-embedded portion of the SRII-HtrII interface, are beginning to produce a clear picture of the signal transfer process. The SR-Htr complexes offer unprecedented opportunities to resolve first examples of the chemistry of signal relay between membrane proteins at the atomic level, which would provide a major contribution to the general understanding of dynamic interactions between integral membrane proteins.


Effect of mutations in SOS genes on UV-induced precise excision of Tn10 in Escherichia coli.
UV treatment increases the frequency of Tn10 precise excision from different sites of the Escherichia coli chromosome. UV induction of Tn10 excision is not evidenced in a lexA3 (ind-) mutant carrying either a recA+ or a recA730 allele. High levels of RecA synthesized by a recA+ gene not repressible by LexA do not relieve the non-inducibility of Tn10 excision in a lexA3 (ind-) background. This indicates that the expression of an SOS gene different from recA is necessary for the induction of Tn10 excision. In contrast to UV induction of point mutations, this induction does not depend on a functional umuC gene since umuC::Tn5 mutants show increased levels of Tn10 excision from leu, thr or gal after irradiation. MucAB+ plasmid pKM101 which renders cells more UV-mutable for point mutations decreases UV-induced Tn10 excision. These results show that UV-induced Tn10 precise excision requires SOS induction and that it involves a pathway different from point mutagenesis.


Systematic Evaluation of Smartphone Applications for the Medical Management of Nephrolithiasis.
Objective: The aim of this study was to comprehensively evaluate the quality of patient-oriented, mobile health applications (apps) available for medical management of kidney stone disease (KSD). Materials and Methods: A systematic search was performed to identify apps from the Apple and Google Play app stores. Eligible apps were divided into two categories: (1) fluid apps and (2) dietary apps. Descriptive characteristics were summarized and app quality was independently evaluated on the basis of technical quality, usability, clinical value, and privacy/security. The validated Mobile App Rating Scale (MARS) was the backbone for the evaluation framework. Results: In total, 31 apps (18 fluid apps and 13 dietary apps) were included. Fluid apps outscore dietary apps in all dimensions of MARS quality. All but one fluid app exceeded the minimum acceptable MARS score of 3.0 vs 46% of dietary management apps. Dietary apps feature less functionality and most are limited to noninteractive informational content. While most data-capturing apps provide a privacy policy and ensure that security measures are taken to protect user identity, only 36% offer users choice or consent for secondary use of their data after data have already been collected. Overall, only one app has undergone any form of real-world clinical investigation. Conclusions: Fluid management apps outperform dietary apps in technical quality, usability, and clinical value at the cost of greater privacy/security risks. Despite not being targeted to patients with KSD, they are higher in most quality measures and offer greater functionality to support long-term disease management compared with dietary apps.


Percutaneous coronary intervention for three vessels of chronic total occlusion complicated with huge left ventricular thrombus.
A 55-year-old woman with symptoms of heart failure appeared to have ischemic cardiomyopathy complicated with 3-vessel chronic total occlusion (CTO), severely deteriorated left ventricular (LV) function, and large LV thrombus. Because of high risk of surgical thrombectomy in addition to coronary artery bypass grafting surgery, we first confirmed significant reduction of LV thrombus and performed percutaneous coronary intervention against the 3-vessel CTO under preceding anticoagulation therapy with warfarin, aspirin, and clopidogrel. By means of antegrade/retrograde approach, we successfully implanted everolimus-eluting stents one by one, leading to complete revascularization, dramatically improved LV function, and disappearance of LV thrombus without post-procedural embolism.


Novel allele of the endothelial nitric oxide synthase gene polymorphism in Caucasian asthmatics.
The purpose of the present study was to investigate an association of the endothelial nitric oxide synthase (eNOS) gene polymorphism with asthma. eNOS intron4 variable number of tandem repeats (27 bp repeats) genotypes were determined in asthma patients and control subjects using microplate PCR. The Caucasian Asthma patients displayed three alleles while Caucasian controls displayed only two alleles. However, African Americans displayed all three alleles in both asthma patients and controls. African American controls displayed significantly higher a allele, significantly lower b allele, as well as c allele than those in Caucasians. This was the first report to show that there was a novel eNOS genotype present in asthmatic patients but not in control subjects in the Caucasian Community. The novel allele which we termed the c allele, could be a significant risk factor in the etiology of asthma in Caucasians. This novel allele could be involved in higher levels of NO in our Caucasian asthmatics.


Modulation of the maitotoxin response by intracellular and extracellular cations.
The aim of the present study was to characterize the role played by intracellular and extracellular calcium and sodium on the maitotoxin (MTX) response in Chinese hamster ovary (CHO) cells. The results presented here indicated that: (1) MTX activates calcium and sodium influx in a concentration-dependent manner; (2) extracellular calcium is required for the sodium influx; (3) removal of the extracellular sodium did not prevent the MTX-induced calcium influx; (4) elevation in the intracellular calcium concentration potentiates the MTX response; and (5) MTX, at the concentrations tested, did not compromise cell viability.


Incorporation of polyunsaturated fatty acids into phospholipids of hemocytes from the tobacco hornworm, Manduca sexta.
We examined the incorporation of four radioactive fatty acids, 18:1n-9, 18:2n-6, 20:4n-6 and 20:5n-3, into cellular lipids of hemocytes from tobacco hornworms, Manduca sexta. Most of the radioactivity associated with 18:1n-9 was recovered from triacylglycerols (TGs), and the radioactivity associated with 18:2n-6 was heavily incorporated into phospholipids (PLs) and TGs. Most of the radioactivity associated with the two eicosanoid-precursor polyunsaturated fatty acids (PUFAs), 20:4n-6 and 20:5n-3, was incorporated into PLs. The incorporated fatty acids were redistributed among the lipid classes during 2 h incubations. The two C20 PUFAs were moved from PLs to TGs. While 18:2n-6 underwent little change, 18:1n-9 was redistributed from TGs to PLs. Within PLs, each of the fatty acids were incorporated into phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PtG) and phosphatidylserine/inositol (PS/PI). The incorporation patterns changed over time, indicating that the incorporated fatty acids were redistributed among the four PL fractions. The radioactivity associated with 18:1n-9 was mostly recovered from the sn-1 position of PC (59%) and PE (83%). Most of the radioactivity associated with 18:2n-6 was found in the sn-2 position of PC (88%) and PE (67%). Over 90% of the radioactivity associated with 20:5n-3 was recovered from the sn-2 position of PC and PE. Incorporation of 20:4n-6 differed from 20:5n-3 because more radioactivity was recovered from the sn-2 position of PC (93%) than PE (69%). These findings are in line with the general background of lipid biochemistry, from which incorporation of 20:4n-6 into PE marks a notable departure: 31% of the radioactivity associated with this acid was recovered from the sn-1 position of PE. These findings indicate that hemocytes from the tobacco hornworm elaborate a fatty acid incorporation system, which exhibits specificity with respect to fatty acid structure and lipid class.


A meta analysis on the utility of Anakinra in severe COVID-19 disease.
The most important presentation of COVID-19 is hyper inflammatory condition and cytokine storm that occurs due to excessive increase of the inflammatory mediators specially, pro-inflammatory interleukins such as IL-1β, IL-6 and tumor necrosis factor-α which have an important role in the cytokine storm pathway. Up till now there is not a definitive treatment for COVID-19 disease, but according to the pathophysiology of the disease, Anakinra (Interleukin- 1 inhibitor) is an adjuvant treatment option in patients with severe COVID-19 by blocking the effect of IL-1. So, we aimed to summarize the studies that evaluated the safety and efficacy of Anakinra in patients diagnosed with COVID-19. We performed a search in PubMed, Cochrane Library, Scopus, and Web of Science (WOS) databases from inception till 7 Jan 2022. Additionally, we searched randomized and non-randomized clinical trials, cohort, case series, case control, case report more than 3 patients which contain confirmed cases of COVID-19 who received Anakinra (Interleukin- 1 inhibitor) for the management of hyper-inflammatory condition associated with COVID-19 disease. A meta-analysis was conducted using review manager 5.4. We included 44 articles in the systematic review. Ultimately, 23 studies were incorporated in the meta-analysis with a total number of 3179 patients. Our analysis showed statistically significant difference in the following outcomes: duration of ICU stays [MD = -0.65, 95% CI (-1.09, -0.03), p = 0.04], the number of patients who needed invasive mechanical ventilation [RR = 0.57, 95% CI (0.39, 0.84), p = 0.004], and number of deaths [RR = 0.80, 95% CI (0.66, 0.99), p = 0.04]. Our analysis showed no statistically significant difference in the following outcomes: length of hospital stays [MD = -0.16, 95% CI (-0.42, 0.11), p = 0.26], oxygen-free days [MD = -0.81, 95% CI (-3.81, 2.20), p = 0.60], and the number of patients who needed non-invasive mechanical ventilation [RR = 1.09, 95% CI (0.47, 2.52), p = 0.84]. Anakinra showed some promising results in important outcomes related to COVID-19 as it significantly reduced the rate of mortality and the need of invasive mechanical ventilation. It should be used in severe cases more than mild and moderate cases to avoid possible immunosuppression complications. Anakinra use is safe in cases of COVID-19 at dose less than 100 mg. Another important outcome was significant reduction is the D-dimer level. Anakinra may be effective in the treatment of specific immunocompromised cases, but it should be used cautiously.


Anti-Neutrophilic Cytoplasmic Antibody (ANCA) Vasculitis Presented as Pulmonary Hemorrhage in a Positive COVID-19 Patient: A Case Report.
Granulomatosis with polyangiitis is a small vessel vasculitis with a wide spectrum of presentation, ranging from limited disease to life-threatening situation such as alveolar hemorrhage. Immunosuppression is the corner stone of the treatment and if left untreated, the death toll increases dramatically. We presented a case of granulomatosis with polyangiitis, presented with alveolar hemorrhage associated with COVID-19 infection. The patient admitted to the intensive care unit, received pulse steroids, plasmapheresis and intravenous immunoglobulin. She was not given further immunosuppression because of the coexisting COVID-19. Up to our knowledge, this is the first reported case of alveolar hemorrhage secondary to granulomatosis with polyangiitis coexisting with COVID-19 infection.


Age-related histopathological findings in temporal arteries.
Giant cell arteritis (GCA) is a systemic vasculitis affecting medium and large arteries in patients aged over 50 years. Involvement of temporal arteries (TA) can lead to complications such as blindness and stroke. While the diagnostic gold standard is temporal artery biopsy (TAB), comorbidities and age-related changes can make interpretation of such specimens difficult. This study aims to establish a baseline of TA changes in subjects without GCA to facilitate the interpretation of TAB. Bilateral TA specimens were collected from 100 consecutive eligible postmortem examinations. Subjects were divided into four age groups and specimens semiquantitatively evaluated for eccentric intimal fibroplasia, disruption and calcification of the internal elastic lamina (IEL), medial attenuation and degree of lymphocytic inflammation of the peri-adventitia, adventitia, media and intima. The individual scores of intimal fibroplasia, IEL disruption and medial attenuation were added to yield a 'combined score (CS)'. Seventy-eight 78 decedents were included in the final analysis following exclusion of 22 individuals for either lack of clinical information or inability to collect TA tissue. A total of 128 temporal artery specimens (50 bilateral from individual decedents, 28 unilateral) were available for examination. Intimal proliferation, IEL loss, IEL calcification and CS increased with age in a statistically significant fashion. Comparison of the oldest age group with the others showed statistically significant differences, although this was not uniformly preserved in comparison between the three youngest groups. Senescent arterial changes and healed GCA exhibit histological similarity and such changes increase proportionally with age. The CS demonstrates significant association with age overall and represents a potential avenue for development to 'normalise' TA biopsies from older individuals.


Differential hypothalamic self-stimulation behaviour in Roman high-avoidance and low-avoidance rats.
Monopolar electrodes were implanted bilaterally and symmetrically into a small well defined region of the posterior lateral hypothalamus in 11 rats of the selected strains Roman high-avoidance (RHA/Verh, 6 animals) and Roman low-avoidance (RLA/Verh, 5 animals). Intracranial electrical stimulation (ICS) was used to study possible strain difference with respect to self-stimulation behaviour. Using a shuttle-box, the experimental set-up allowed measurements of the animal's preference, aversion, neutrality or ambivalence towards the stimulus. For both rat strains 10 electrode sites were tested with identical stimulation currents (100-600 microA in increments of 100 microA). Significant differences between the selected strains were found for preference (self-stimulation) and aversion: RLA/Verh-rats showed self-stimulation at lower stimulation currents than RHA/Verh-rats. At higher currents, both strains showed similar self-stimulation performance. RLA/Verh-rats escaped more often from ICS yielding self-stimulation at five of the six current levels. It is concluded that, in comparison with the RHA strain, RLA/Verh-rats are more sensitive to aversive effect of lateral hypothalamic stimulation. This seems to be associated with an increased sensitivity to the rewarding effects of such stimulation.


Role of Bcl-2 family members in immunity and disease.
The different members of the Bcl-2 family are essential regulators of programmed cell death. These different members share one or more Bcl-2 homology domains, required for their ability to regulate each other. In this review, we describe current knowledge of the functions of different Bcl-2 members and their potential roles in disease and immunity.


Hypertensive Emergency in Heart Failure: Trends, Risk factors and Outcomes from a Nationwide Analysis 2005-2014.
Heart failure (HF) patients may be susceptible to complications of hypertensive emergency (HTNE). Large registries have found that these patients are not on optimal antihypertensive therapy. To date, little investigation has been done on HF patients with HTNE, and their clinical risk factors/outcomes have not been well defined. We reviewed the National Inpatient Sample database to collect data on HF patient hospitalizations from 2005 to 2014. HF patients with and without a primary diagnosis of HTNE were included in the analysis. Risk factors and outcomes of HF patients with a primary diagnosis of HTNE were compared to those without HTNE. HF patients with a primary diagnosis of shock of any etiology were excluded. Of 8,265,792 patients hospitalized with HF, 39,170 (0.5%) had HTNE. There was a threefold increase in the incidence of HTNE in HF patients over a 10-year period. The increase was more evident in females, Blacks and those < 40 years of age. There was also an increasing trend in modifiable risk factors. HF patients with HTNE had significantly higher major in-hospital complications compared to those without HTNE. However, this association did not reflect an increase in short-term in-hospital mortality, irrespective of age. HF patients with HTNE represent a unique population that requires a different approach to treatment. Further research is needed to identify barriers preventing adequate therapy of hypertension and other modifiable risk factors in HF patients and assess their effects on long-term outcomes.


Phospholipase A2 inhibitors. Differential inhibition of fatty acid acylation into kidney lipids by mepacrine and p-bromophenacyl bromide.
Mepacrine and p-bromophenacyl bromide, in addition to their inhibitory effect on lipolysis, are also potent inhibitors of fatty acid acylation into renal medullary lipids. Significant qualitative and quantitative differences in the inhibition by the two drugs were seen. p-Bromophenacyl bromide exerted a non-selective effect inhibiting the incorporation of saturated and unsaturated fatty acids into all phospholipid classes and triacylglycerols. In contrast, mepacrine selectively inhibited the incorporation of both saturated and unsaturated acids into phosphatidylcholine, phosphatidylethanolamine and triglycerides, and concurrently markedly enhanced their incorporation into phosphatidylinositol. Quantitative analysis of these mepacrine effects, together with the known inhibitory effects of this compound on phospholipase A2 and phosphatidylinositol-specific phospholipase C, suggests that mepacrine also inhibits phosphatidic acid phosphatase, thereby shunting the flux of phosphatidic acid away from diglyceride formation and into synthesis of phosphatidylinositol.


Experimental schizophrenia models in rodents established with inflammatory agents and cytokines.
Immune inflammatory processes in prenatal and perinatal stages are suggested to play crucial roles in the vulnerability to schizophrenia. Based upon this immune inflammatory hypothesis for schizophrenia, we have established animal models for this illness by subcutaneously administering cytokines or proinflammatory agents to rodent neonates. These models exhibit various schizophrenia-like behavioral abnormalities after puberty, most of which are sensitive to various antipsychotics. The experimental procedures are all simple and easily utilized by researchers unfamiliar with these models. The behavioral changes are reproducible and remarkable but do not accompany learning deficits. The molecular and cellular targets of these agents have also been investigated and partially characterized, such as the cortical GABAergic system, midbrain dopaminergic system and hippocampal glutamate system. In this chapter, we introduce the details of the procedure and discuss the potential application of these animal models to drug development for schizophrenia.


Tranexamic acid accelerates skin barrier recovery and upregulates occludin in damaged skin.
Tranexamic acid (TA) is a traditional plasmin inhibitor, and its role in the renovation of damaged skin has become the topic of a lot of research. The aim of this study is to determine whether TA could repair the skin barrier by means of tight intercellular junctions. Two kinds of damaged skin models were set up and subjected to repeated application of sodium lauryl sulfate and irradiation of ultraviolet B. Through bioengineering technology and immunohistochemistry tests, TA-induced changes in skin were detected. After 1, 3, 7, and 14 days of application, TA can significantly accelerate barrier recovery and decrease the melanin index values of ultraviolet B irritation skin. The mean optic density of occludin from TA treatment is higher than from self-repair. These experiments suggest that TA can accelerate skin barrier recovery and upregulate occludin induced by physicochemical damages of human skin, but it is advisable to perform more research on the upregulation of occludin in molecular mechanism in the future.


Investigation of active pharmaceutical ingredient loss in pharmaceutical compounding of capsules.
Pharmaceutical compounding of capsules is still an important corner stone in today's health care. It allows for a more patient specific treatment plan as opposed to the "one size fits all"-approach, used by the pharmaceutical industry when producing fixed dose finished drug products. However, loss of active pharmaceutical ingredient (API) powder during pharmaceutical capsule compounding can lead to under-dosed finished drug products and annul the beneficiary therapeutic effects for the patient. The amount and location of API loss was experimentally determined during capsule compounding of five different preparations: 10 and 20mg hydrocortisone capsules, 4mg triamcinolone capsules and 0.25mg dexamethasone capsules, using a 10% m/m self-made or commercial trituration. The total API amount present in the five capsule preparations varied between 90.8% and 96.6%, demonstrating that for certain preparations, significant API mass loss occurred during the pharmaceutical compounding of capsules. Swabbing results of the different compounding equipment and working areas indicated the mortar surface as the largest API loss location. An agate mortar accounted for the least amount of API loss, whereas an extensively used porcelain mortar accounted for the highest amount of API loss. Optical microscopy and roughness (Ra) determination by profilometry of the different mortar surfaces revealed a significant influence of the mortar surface wear and tear on the observed API loss. This observation can be explained by physical deformation, or scratch formation, of the relatively soft porcelain mortar surface, in which the API particles can become adsorbed. Furthermore, a small effect of the capsulation device material on the API loss was also observed. The presence of a chemical molecule effect on the API loss was demonstrated through data mining using a set of assay results containing 17 different molecules and 1922 assay values. The 17 median assay values were modeled in function of corresponding molecular descriptors, using stepwise multiple linear regression. The obtained MLR model, containing RDF060m, R6e(+) and R3m(+) variables, explained 92.5% of the observed variability between the 17 median assay values.


Mumic acids A-E: new diterpenoids from mumiyo.
Five new diterpenoids belonging to labdane and isopimarane skeletons, mumic acids A-E (1-5), have been isolated from mumiyo. Their structures and absolute configurations were elucidated on the basis of spectroscopic data and chemical derivatization.


The reliability of the detection of an early diastolic notch with uterine artery Doppler velocimetry.
This study evaluates the ability of two reviewers to detect independently an early diastolic notch in 1371 uterine artery Doppler velocity waveform recordings. Agreement between the two reviewers for the detection of uterine artery notching was assessed by using the Kappa statistic. The inter-rater reliability for the detection of unilateral notching was 0.75 (95% CI 0.70-0.80), whereas that for the presence or absence of bilateral notching was 0.66 (95% CI 0.60-0.71). The results suggest that there was good reviewer agreement for the presence or absence of a notch on uterine artery Doppler velocimetry.


Effect of desflurane/remifentanil anaesthesia on glucose metabolism during surgery: a comparison with desflurane/epidural anaesthesia.
The aim of this study was to investigate the effect of general anaesthesia combined with remifentanil or epidural blockade on glucose metabolism during surgery. We randomly assigned patients undergoing elective colorectal surgery to receive either desflurane anaesthesia supplemented with intravenous remifentanil (n = 7) or desflurane anaesthesia supplemented with epidural bupivacaine (n = 7). Plasma concentrations of glucose, lactate, free fatty acids (FFA), insulin, glucagon and cortisol were measured before and after 2 h of surgery. Pre- and intraoperative whole body glucose production and glucose clearance, an indicator of glucose uptake, were determined by an isotope dilution technique using [6,6-2H2]glucose. In both groups intraoperative glucose production ( P< 0.05) and uptake ( P< 0.05) decreased. Plasma glucose concentrations ( P< 0.05) increased during surgery but did not exceed the normal range (remifentanil group: 5.7 +/- 0.7 mmol l-1, epidural group: 5.8 +/- 0.4 mmol l-1). The plasma concentrations of lactate, FFA, insulin and glucagon remained unchanged during the operation. The plasma cortisol concentration in both groups increased intraoperatively (P< 0.05). Both desflurane/remifentanil and desflurane/epidural anaesthesia decrease the intraoperative rate of whole body glucose production, thereby attenuating the hyperglycaemic response to colorectal surgery.


Human Milk Lipids Induce Important Metabolic and Epigenetic Changes in Neonates.
Lipids are a major source of energy during the fetal/neonatal period. Most are received from the mother, transplacentally during the intrauterine period or via maternal milk after birth. However, in addition to the known nutritional roles, lipids are now known to bind a variety of cellular receptors to regulate specific patterns in metabolism and gene expression. The expression of these receptors is regulated by various genetic and environmental stimuli, and ligation can activate positive-feedback loops in the expression and the activity of downstream signaling pathways. The authors summarize the role of lipid ligands, cognate receptors, epigenetic regulation, and downstream signaling.


Extremes in second molar eruption in two oriental patients.
Two cases are presented where the maxillary second molars were just erupting. The first was a Chinese boy of 8 years 2 months and the other, a Chinese boy of 17 years 4 months. These extremes in eruption time are unusual and do not appear to have been reported in the literature.


The prevalence and clinical relevance of hyperhomocysteinemia suggesting vitamin B12 deficiency in presumed healthy infants.
Previous studies have demonstrated a high prevalence of biochemical vitamin B12 deficiency in infants in Norway. Increased total homocysteine (tHcy) is the most important marker of B12 deficiency in infants. There is a need to evaluate its clinical relevance. To investigate the prevalence of hyperhomocysteinemia (S-tHcy > 8 μmol/L) suggestive of suboptimal B12 status and the prevalence of clinically relevant hyperhomocysteinemia in presumed healthy infants in Norway. Further, to evaluate risk factors, presence of symptoms and psychomotor development in these children. In a prospective study we clinically examined 252 infants aged 3-7 months using standardized neurological and psychomotor tests prior to analyzing biochemical B12 deficiency markers in 250 infants. Twenty-five of 250 (10%) infants had hyperhomocysteinemia combined with clinically relevant symptoms suggestive of B12 deficiency. Hyperhomocysteinemia was associated with tremor, excessive sleep, and sub-normal scores in the fine motor section of the Ages and Stages Questionnaire. One-hundred and fourteen of 250 (46%) infants had hyperhomocysteinemia. Multiple regression analysis showed months of infant formula use as the strongest negative predictor for hyperhomocysteinemia. We have demonstrated associations between symptoms suggestive of infant B12 deficiency and increased levels of tHcy in presumed healthy infants The combination of hyperhomocysteinemia and associated relevant symptoms suggestive of B12 deficiency was a common finding, albeit most infants with hyperhomocysteinemia did not show symptoms.


Quantification of the electrochemical proton gradient and activation of ATP synthase in leaves.
We have developed a new method to quantify the transmembrane electrochemical proton gradient present in chloroplasts of dark-adapted leaves. When a leaf is illuminated by a short pulse of intense light, we observed that the light-induced membrane potential changes, measured by the difference of absorption (520 nm-546 nm), reach a maximum value (approximately 190 mV) determined by ion leaks that occur above a threshold level of the electrochemical proton gradient. After the light-pulse, the decay of the membrane potential follows a multiphasic kinetics. A marked slowdown of the rate of membrane potential decay occurs approximately 100 ms after the light-pulse, which has been previously interpreted as reflecting the switch from an activated to an inactivated state of the ATP synthase (Junge, W., Rumberg, B. and Schröder, H., Eur. J. Biochem. 14 (1970) 575-581). This transition occurs at approximately 110 mV, thereby providing a second reference level. On this basis, we have estimated the Delta micro (H(+)) level that pre-exists in the dark. Depending upon the physiological state of the leaf, this level varies from 40 to 70 mV. In the dark, the Delta micro (H(+)) collapses upon addition of inhibitors of the respiratory chain, thus showing that it results from the hydrolysis of ATP of mitochondrial origin. Illumination of the leaf for a period longer than several seconds induces a long-lived Delta micro (H(+)) increase (up to approximately 150 mV) that reflects the light-induced increase in ATP concentration. Following the illumination, Delta micro (H(+)) relaxes to its dark-adapted value according a multiphasic kinetics that is completed in more than 1 h. In mature leaf, the deactivation of the Benson-Calvin cycle follows similar kinetics as Delta micro (H(+)) decay, showing that its state of activation is mainly controlled by ATP concentration.


Clinical and biochemical characteristics of patients with ornithine transcarbamylase deficiency.
Ornithine transcarbamylase deficiency (OTCD) is pleomorphic congenital hyperammonemia, in which the prognosis of the patient is determined both by genotype and environmental factors. This study investigated the clinical and biochemical characteristics of OTCD patients with different prognosis. Of 35 OTCD patients, six males deceased at the first disease-onset, 17 males survived and had controllable ammonia levels after treatment, and 12 females survived through the first disease-onset but had intractable hyperammonemia and high mortality. Fasting blood samples from patients collected at three disease stages were used for the analysis of amino acid (AA) profile, acylcarnitine profile, and micronutrients. Differences in profiles between patients and healthy controls and within patient groups were studied. All OTCD patients had accumulation of glutamine, homocitrulline, lysine, glutamate, cystathionine, and pipecolic acid, as well as deficiency of citrulline, tryptophan, threonine, and carnitine. For male non-survivors, most other AAs and long-chain acylcarnitines were elevated at disease onset, of which the levels of creatine, N-acetylaspartic acid, and homoarginine were remarkably high. Male survivors and female patients had most other AAs at low to normal levels. Compared with male survivors, female patients had much lower protein-intolerance, as indicated by significantly lower levels of protein consumption indicators, including essential AAs, 1-methylhistidine, acylcarnitines et al., but high levels of ammonia. Female patients still had significantly higher levels of citrulline, homocitrulline, and citrulline/arginine compared to male survivors. Unique profiles were observed in each group of OTCD patients, indicating specific physiological changes that happened to them.


Nicotine administration decreases nitric oxide synthase expression in the hypothalamus of food-deprived rats.
Effect of nicotine on nitric oxide synthase (NOS) expression in various hypothalamic regions was investigated in rats via nicotineamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry. Sprague-Dawley rats were divided into the fed group, the fed and nicotine-treated group, the food-deprived group, and the food-deprived and nicotine-treated group. The fed groups received abundant food and water, while food was withheld from the food-deprived groups for 48 h. The nicotine-treated groups were injected with nicotine. Following food deprivation, enhanced NAPDH-d expression was detected in the paraventricular nucleus, ventromedial hypothalamic nucleus, and lateral hypothalamic area of the hypothalamus. Nicotine administration to the food-deprived rats resulted in decreased NADPH-d positivity. The present results indicate that nicotine administration is effective in limiting the enhancement in NOS expression following food restriction.


Chemical constituents from Sonneratia ovata Backer and their in vitro cytotoxicity and acetylcholinesterase inhibitory activities.
Sonneratia ovata Backer, Sonneratiaceae, is a widespread plant in mangrove forests in Vietnam, Cambodia, Thailand, Indonesia. Sonneratia ovata's chemical composition remains mostly unknown. Therefore, we now report on the structural elucidation of three new phenolics, sonnerphenolic A (1), sonnerphenolic B (2), and sonnerphenolic C (23), a new cerebroside, sonnercerebroside (3) together with nineteen known compounds, including nine lignans (5-13), two steroids (14, 15), two triterpenoids (16, 17), three gallic acid derivatives (18-20), two phenolic derivatives (4, 22) and a 1-O-benzyl-β-d-glucopyranose (21) isolated from the leaves of Sonneratia ovata. Their chemical structures were established by spectroscopic data, as well as high resolution mass spectra and comparison with literature data. The in vitro acetylcholinesterase (AChE) inhibition and cytotoxic activities against HeLa (human epithelial carcinoma), NCI-H460 (human lung cancer), MCF-7 (human breast cancer) cancer cell lines and PHF (primary human fibroblast) cell were evaluated on some extracts and purified compounds at a concentration of 100 μg/mL. Compounds (5, 6, 23) exhibited cytotoxicity against the MCF-7 cell line with the IC50 values of 146.9±9.0, 114.5±7.2, and 112.8±9.4 μM, respectively, while they showed nontoxic with the normal cell (PHF) with IC50s >277 μM. Among 15 tested compounds, (S)-rhodolatouchol (22) showed inhibition against AChE with an IC50 value of 96.1±14.5 μM.


Outcome evaluation of minimally invasive surgery versus extensile lateral approach in management of displaced intra-articular calcaneal fractures: A randomised control trial.
Minimally invasive surgery has a significant and evolving role in the treatment of DIACFs, but there is limited literature on this topic. To compare the clinico-radiological outcomes of DIACFs fixed with MIS technique with ORIF. This randomised control trial (RCT) included 21 closed Sanders type 2 and 3 DIACFs which were selected from 70 who presented. Extensile lateral approach was used in the ORIF group; while MIS techniques included either percutaneous reduction or small incisions with indirect fragment manipulation. AOFAS hindfoot score, pre-operative and postoperative radiology. There were 9 fractures in the Extensile group and 12 in the MIS group, with both groups having comparable demography, fracture classification, surgery delay & initial radiology. Bohler's angle improved after surgery by an average of 18.44 with ORIF and 14.67 with MIS (p=0.28). Mean AOFAS in the MIS group was 82.58 (66.67% good and 33.33% fair) and was 89.56 (44.44% excellent, 55.56% good, 0 fair/poor) with ORIF (p=0.034). Two patients who underwent ORIF had a major complication (one deep infection and Sural nerve injury each) while none in the MIS group. MIS methods minimized soft tissue complications and achieved comparable radiological reductions but clinical outcomes were poorer, with percutaneous methods having the worst outcomes.


Partial urogenital mobilization: a limited proximal dissection.
The treatment of urogenital sinus malformations is complex and controversial. Despite numerous and significant contemporary surgical advances, the dissection of the urogenital sinus remains technically challenging. Based on total urogenital mobilization, we describe a technique whereby this dissection is limited to the pubourethral ligament. Our short-term results with partial urogenital mobilization (PUM) performed on 15 patients are retrospectively reviewed. There were no intraoperative complications and the short-term cosmetic results are excellent. No patients have developed voiding dysfunction or urinary tract complications. While total urogenital mobilization is a very effective procedure, we believe that the PUM approach limits potential morbidity in the reconstruction of these complex problems.


Utility of a public-available artificial intelligence in diagnosis of polypoidal choroidal vasculopathy.
To investigate the feasibility of training an artificial intelligence (AI) on a public-available AI platform to diagnose polypoidal choroidal vasculopathy (PCV) using indocyanine green angiography (ICGA). Two methods using AI models were trained by a data set including 430 ICGA images of normal, neovascular age-related macular degeneration (nvAMD), and PCV eyes on a public-available AI platform. The one-step method distinguished normal, nvAMD, and PCV images simultaneously. The two-step method identifies normal and abnormal ICGA images at the first step and diagnoses PCV from the abnormal ICGA images at the second step. The method with higher performance was used to compare with retinal specialists and ophthalmologic residents on the performance of diagnosing PCV. The two-step method had better performance, in which the precision was 0.911 and the recall was 0.911 at the first step, and the precision was 0.783, and the recall was 0.783 at the second step. For the test data set, the two-step method distinguished normal and abnormal images with an accuracy of 1 and diagnosed PCV with an accuracy of 0.83, which was comparable to retinal specialists and superior to ophthalmologic residents. In this evaluation of ICGA images from normal, nvAMD, and PCV eyes, the models trained on a public-available AI platform had comparable performance to retinal specialists for diagnosing PCV. The utility of public-available AI platform might help everyone including ophthalmologists who had no AI-related resources, especially those in less developed areas, for future studies.


Brain-specific Pd1 deficiency leads to cortical neurogenesis defects and depressive-like behaviors in mice.
Embryonic neurogenesis is tightly regulated by multiple factors to ensure the precise development of the cortex. Deficiency in neurogenesis may result in behavioral abnormalities. Pd1 is a well-known inhibitory immune molecule, but its function in brain development remains unknown. Here, we find brain specific deletion of Pd1 results in abnormal cortical neurogenesis, including enhanced proliferation of neural progenitors and reduced neuronal differentiation. In addition, neurons in Pd1 knockout mice exhibit abnormal morphology, both the total length and the number of primary dendrites were reduced. Moreover, Pd1cKO mice exhibit depressive-like behaviors, including immobility, despair, and anhedonia. Mechanistically, Pd1 regulates embryonic neurogenesis by targeting Pax3 through the β-catenin signaling pathway. The constitutive expression of Pax3 partly rescues the deficiency of neurogenesis in the Pd1 deleted embryonic brain. Besides, the administration of β-catenin inhibitor, XAV939, not only rescues abnormal brain development but also ameliorates depressive-like behaviors in Pd1cKO mice. Simultaneously, Pd1 plays a similar role in human neural progenitor cells (hNPCs) proliferation and differentiation. Taken together, our findings reveal the critical role and regulatory mechanism of Pd1 in embryonic neurogenesis and behavioral modulation, which could contribute to understanding immune molecules in brain development.


Differential effects of combinations of phospholipase A2 and phospholipase C on the activity of rat epididymal nuclear and microsomal 4-ene steroid 5 alpha-reductase.
Epididymal 4-ene steroid 5 alpha-reductase converts testosterone to 5 alpha-dihydrotestosterone. The enzyme is localized to the nuclear and microsomal fractions, and the activity can be altered by modifying the phospholipids in the membrane environment. To investigate the membrane dependence of 4-ene steroid 5 alpha-reductase, we have treated nuclear and microsomal membranes with combinations of phospholipase A2 and phospholipase C, and examined the effects on 4-ene steroid 5 alpha-reductase activity. Sequential addition of phospholipase A2 and phospholipase C to the nuclear fraction, reduced the 4-ene steroid 5 alpha-reductase activity to approx 25% of the control level. Neither the nature of the phospholipase, nor the sequence of addition altered the inhibition. When both phospholipases were added simultaneously, nuclear 4-ene steroid 5 alpha-reductase activity was inhibited in a linear fashion, and in tests for cooperativity, the effects of phospholipase A2 and phospholipase C were clearly additive. The microsomal enzyme responded differently to sequential phospholipase treatments; if phospholipase A2 was followed by phospholipase C, or phospholipase C followed by phospholipase A2, the 4-ene steroid 5 alpha-reductase activity was, respectively, 13 and 27% of the control. In contrast, sequential addition of the same phospholipase reduced the activity of 4-ene steroid 5 alpha-reductase to approx 40% of the control level. Furthermore, simultaneous addition of phospholipase A2 and phospholipase C to the microsomal fraction, resulted in non-linearity of 4-ene steroid 5 alpha-reductase activity with time, whereas when added individually, linearity of 4-ene steroid 5 alpha-reductase was maintained. Consequently, it was not possible to test for cooperative effects of phospholipases on the microsomal 4-ene steroid 5 alpha-reductase. These findings suggest that for the nuclear 4-ene steroid 5 alpha-reductase, the polar and non-polar regions of the membrane environment have similar functions, which are most likely involved in the maintenance of the structural integrity of the enzyme. For the microsomal enzyme, the polar and non-polar regions of the membrane appear to have different functions, not only for the maintenance of enzyme integrity, but also in the mechanism at the active site.


Solid phase microextraction of polycyclic aromatic hydrocarbons from water samples by a fiber coated with covalent organic framework modified graphitic carbon nitride.
A covalent organic framework modified graphitic carbon nitride (g-C3N4@TpBD) was synthesized by modifying the graphitic carbon nitride (g-C3N4) with a covalent organic framework (COF-TpBD). The synthesis conditions including the mass ratio between g-C3N4 and benzidine (BD), solvent type, reaction temperature and reaction time were optimized. Under the optimal synthetic conditions, a novel spiny dendritic g-C3N4@TpBD adsorbent was obtained. The g-C3N4@TpBD was then coated on stainless-steel wire by sol-gel technique and the coated fiber was used for the solid phase microextraction of polycyclic aromatic hydrocarbons prior to gas chromatography-mass spectrometric detection. The established method was successfully applied to determine eight PAHs in six environmental water samples. Under the optimal extraction conditions, a wide linear quantification range for the analytes was obtained from 0.07 to 60.0 ng mL-1 with the coefficients of determination varying from 0.9979 to 0.9998, and the limits of detection (S/N = 3) ranged from 0.02 to 0.05 ng mL-1. The relative recoveries of the analytes for the six environmental water samples at the spiked concentrations of 0.2, 0.5, 3.0 and 30.0 ng mL-1 were between 83.6% and 118% with the relative standard deviations ranging from 2.4% to 11.3%.


Mortality in Patients With Chronic Renal Disease Without Health Insurance in Mexico: Opportunities for a National Renal Health Policy.
Despite a systematic increase in the coverage of patients with end-stage renal disease (ESRD) who have received dialytic therapies and transplantation over the past 2 decades, the Mexican health system currently still does not have a program to provide full coverage of ESRD. Our aim was to analyze mortality in patients with ESRD without health insurance. This was a prospective cohort study of 850 patients with advanced chronic kidney disease (CKD). Risk factors associated with death were calculated using a Cox's proportional hazards model. We used the statistical package SPSS version 22.0 for data analysis. The mean age of patients was 44.8 ± 17.2 years old. At the time of hospital admission, 87.6% of the population did not have a social security program to cover the cost of renal replacement treatment, and 91.3% of families had an income below US$300 per month. During the 3 years of the study, 28.8% of the cohort patients were enrolled in 1 of Mexico's social security programs. The 3-year mortality rate was of 56.7% among patients without access to health insurance, in contrast to 38.2% of patients who had access to a social security program that provided access to renal replacement therapy (P < 0.001). Risk factor analysis revealed that not having health insurance increased mortality (risk ratio: 2.64, 95% confidence intervals: 1.84-3.79; P = 0.001). Mexico needs a coordinated National Kidney Health and Treatment Program. A program of this nature should provide the basis for an appropriate educational and intervention strategy for early detection, prevention, and treatment of patients with advanced chronic kidney disease.


Reversible cerebral vasoconstriction syndrome after levonorgestrel-releasing intrauterine device placement-Case report.
Reversible cerebral vasoconstriction syndrome (RCVS) is a disease characterized by reversible multifocal narrowing of the cerebral arteries with clinical manifestations that typically include thunderclap headache and occasionally brain edema, stroke, or seizure. The exact pathophysiology of RCVS is not well known. A 46-year-old female with history of episodic migraine presented with 1-month duration of worsening headaches that had become more severe over the past 2 weeks. The headaches were episodic and thunderclap in onset and aggravated by physical exertion or emotional situations. A neurological examination was unremarkable including initial head computed tomography (CT). A CT angiogram of the head showed multifocal stenosis in the right anterior cerebral artery, bilateral middle cerebral arteries, and right posterior cerebral artery. Cerebral angiogram confirmed the CT angiogram findings. A repeated CT angiogram a few days later showed improvement in the multifocal cerebral arterial stenosis. Lumbar puncture and autoimmune workup were not suggestive of neuroinflammatory etiology. She had one generalized tonic-clonic seizure during her second day of hospitalization. The patient's thunderclap onset headaches resolved in 1 week after she was managed with blood pressure control and pain medication. She denied any illicit drug use or any new medications other than the placement of a levonorgestrel-releasing intrauterine device (IUD) about 6 weeks prior to her presentation. Our case suggests a possible link between RCVS and levonorgestrel-releasing IUDs.


Modulation of thyroid-specific gene expression in normal and nodular human thyroid tissues from adults: an in vivo effect of thyrotropin.
Evidence from in vitro studies or animal models has shown that TSH affects thyrocytes by thyroid-specific expression modulation. The objective of our study was to analyze the role of TSH in human thyroid gene expression in vivo. Thirty-nine normal thyroid tissues were collected at the same center. Patients were divided into two groups based on serum TSH levels: 17 with normal TSH levels (1-4 mU/liter; group 1) and 22 with TSH levels below 0.5 mU/liter (group 2). Group 2 underwent thyroidectomy after suppressive L-T4 therapy. mRNA levels of thyroid genes such as sodium/iodide symporter (NIS), apical iodide transporter, pendrin, thyroglobulin, thyroperoxidase, TSH receptor, paired box transcription factor 8, and thyroid transcription factor-1 were evaluated by quantitative PCR. The reduction of TSH stimulation causes decreases in NIS and apical iodide transporter gene expression in normal tissues and more limited reductions in thyroglobulin, thyroperoxidase, and paired box transcription factor 8, but it has no significant effect on TSH receptor, pendrin, or thyroid transcription factor-1. Comparison of NIS levels in normal and nodular tissues from the same patient confirmed that it is differentially expressed in nodules only in the presence of normal TSH (P < 0.01). In patients with suppressed TSH, nodular NIS levels were similar to those in normal tissues. Our data represent the first demonstration in human thyroid tissues that TSH contributes to the regulation of thyrocyte differentiation by modulating thyroid gene levels. It exerts a particularly important effect on the transcription of NIS, which becomes very low after prolonged TSH suppression.


Overexpression of the Drosophila ATR homologous checkpoint kinase Mei-41 induces a G2/M checkpoint in Drosophila imaginal tissue.
DNA damage generally results in the activation of ATM/ATR kinases and the downstream checkpoint kinases Chk1/Chk2. In Drosophila melanogaster, the ATR homologue meiotic 41 (mei-41) is pivotal to DNA damage repair and cell cycle checkpoint signalling. Although various mei-41 mutant alleles have been analyzed in the past, no gain-of-function allele is yet available. To fill this gap, we have generated transgenic flies allowing temporal and tissue-specific induction of mei-41. Overexpression of mei-41 in wing and eye anlagen affects proliferation and a G2/M checkpoint even in the absence of genomic stress. Similar consequences were observed following the overexpression of the downstream kinase Grapes (Grp) but not of Loki (Lok), encoding the respective Drosophila Chk1 and Chk2 homologues, in agreement with their previously reported activities. Moreover, we show that irradiation induced cell cycle arrest was prolonged in the presence of ectopic mei-41 expression. Similar to irradiation stress, mei-41 triggered the occurrence of a slower migrating form of Grp, implying specific phosphorylation of Grp in response to either signal. Using a p53R-GFP biosensor, we further show that overexpression of mei-41 was sufficient to elicit a robust p53 activation in vivo. We conclude that overexpression of the Drosophila ATR homologue mei-41 elicits an effectual DNA damage response irrespective of irradiation.


Spontaneous spinal epidural hematoma in an untreated hypertensive patient.
An insufficiently treated hypertensive woman developed severe neck pain, followed by a partial Brown-Sequard syndrome, due to a spontaneous spinal epidural hematoma compressing the cervical spinal cord at the levels C4-C7. We discuss the diagnostic difficulties in the acute setting and underline the importance of a careful and detailed neurological evaluation in all patients with any kind of acute neurological deficit. Application of modern imaging techniques in such selected acute cases is helpful in order to establish a diagnosis and initiate therapy. Although rare, clinicians should keep spinal epidural hematoma in mind and consider it as a highly uncommon, yet possible, complication of untreated hypertension.


Cesarean delivery and respiratory distress syndrome: does labor make a difference?
The purpose of this study was to determine if cesarean delivery is a risk factor for respiratory distress syndrome (RDS) and if this risk is modified by labor before cesarean. This population-based case-control study compared 4778 cases of RDS to 5 times as many controls. Unadjusted, cesarean delivery was associated with RDS, odds ratio (OR) 3.5 (95% CI 3.2-3.8). After controlling for potential confounding variables, cesarean remained an independent risk factor, OR 2.3 (95% CI 2.1-2.6). Labor modified this risk significantly (P = .02)--with labor, cesarean delivery had an OR of 1.9 (95% CI 2.2-2.9), without labor, the OR was 2.6 (95% CI 1.3-2.8). Cesarean delivery was an independent risk factor for RDS. The risk was reduced with labor before cesarean, but still elevated. This supports the importance of being certain of fetal lung maturity before cesarean delivery, particularly when done before labor.


Protection afforded by allopurinol in the first 24 hours of coronary occlusion is diminished after 48 hours.
Experiments were performed to test whether the reduction in infarct size afforded by allopurinol following 24 h of permanent coronary artery occlusion is sustained over the subsequent 24 h. A dog's coronary artery was occluded with an embolus followed by injection of radiomicrospheres into the left ventricle to mark the ischemic region and to measure regional blood flow. Dogs were sacrificed either 24 h or 48 hours after embolization. The infarcts were delineated by failure to stain with triphenyl tetrazolium chloride and the ischemic zones were visualized by autoradiography of the heart slices. Dogs in the treatment groups received 600 mg of allopurinol PO 18 h before surgery, and a 10 mg/kg IV bolus 15 minutes before embolization followed by constant IV infusion of 55 mg/kg/24 h until sacrifice. A close correlation in the control animals between the percent of the ischemic zone which infarcted and collateral blood flow was used to predict a nonintervention infarct size in each treatment animal. Allopurinol treatment caused 17.9 +/- 3.3% less of the risk zone to be tetrazolium negative after 24 hours of ischemia than that seen in untreated animals. Less allopurinol induced salvage was observed in the 48 hour drug group with only a 11.1 +/- 3.3% limitation in infarct size. Furthermore, the effect was inconsistent at 48 h with only 2 dogs showing salvage. We conclude that allopurinol delays but does not prevent infarction in the permanent occlusion model.


[Determination of sex differentiated ossification patterns of costicartilage pairs II to VI: post mortem study of radiograms of the anterior chest wall].
On the basis of widely laterally resected anterior chest walls, ossification specimens of femal (n = 95) and male (n = 112) individuals of different age have been examined. An unexpected high significance of the already described sex-dimorphism could be secured between the 20th and 50th year of age. The estimated epiphenomenological changes impress as a female calcification type with centrally in the costal cartilage situated ossifications and as a male calcification type with sheath-like calcifications situated at the cranial and caudal edge of the costicartilage. Isolatedly observed contra-sexual ossifications specimens are expressively hinted at.


Donor Heterogeneity in the Human Macrophage Response to a Biomaterial Under Hyperglycemia In Vitro.
Macrophages have a commanding role in scaffold-driven in situ tissue regeneration. Depending on their polarization state, macrophages mediate the formation and remodeling of new tissue by secreting growth factors and cytokines. Therefore, successful outcomes of material-driven in situ tissue vascular tissue engineering depend largely on the immuno-regenerative potential of the recipient. A large cohort of patients requiring vascular replacements suffers from systemic multifactorial diseases, such as diabetes, which gives rise to a hyperglycemic and aggressive oxidative inflammatory environment that is hypothesized to hamper a well-balanced regenerative process. Here, we aimed at fundamentally exploring the effects of hyperglycemia, as one of the hallmarks of diabetes, on the macrophage response to three-dimensional (3D) electrospun synthetic biomaterials for in situ tissue engineering, in terms of inflammatory profile and tissue regenerative capacity. To simulate the early phases of the in situ regenerative cascade, we used a bottom-up in vitro approach. Primary human macrophages (n = 8 donors) were seeded in two-dimensional (2D) culture wells and polarized to pro-inflammatory M1 and anti-inflammatory M2 phenotype in normoglycemic (5.5 mM glucose), hyperglycemic (25 mM), and osmotic control (OC) conditions (5.5 mM glucose, 19.5 mM mannitol). Unpolarized macrophages and (myo)fibroblasts were seeded in mono- or co-culture in a 3D electrospun resorbable polycaprolactone bisurea scaffold and exposed to normoglycemic, hyperglycemic, and OC conditions. The results showed that macrophage polarization by biochemical stimuli was effective under all glycemic conditions and that the polarization states dictated expression of the receptors SCL2A1 (glucose transporter 1) and CD36 (fatty acid transporter). In 3D, the macrophage response to hyperglycemic conditions was strongly donor-dependent in terms of phenotype, cytokine secretion profile, and metabolic receptor expression. When co-cultured with (myo)fibroblasts, hyperglycemic conditions led to an increased expression of fibrogenic markers (ACTA2, COL1, COL3, IL-1β). Together, these findings show that the hyperglycemic and hyperosmotic conditions may, indeed, influence the process of macrophage-driven in situ tissue engineering, and that the extent of this is likely to be patient-specific. Impact Statement Success or failure of cell-free bioresorbable in situ tissue-engineered vascular grafts hinges around the immuno-regenerative response of the recipient. Most patients requiring blood vessel replacements suffer from additional multifactorial diseases, such as diabetes, which may compromise their intrinsic regenerative potential. In this study, we used a bottom-up approach to study the effects of hyperglycemia, a hallmark of diabetes, on important phases in the in situ regenerative cascade, such as macrophage polarization and macrophage-myofibroblast crosstalk. The results demonstrate a relatively large donor-to-donor variation, which stresses the importance of taking scaffold-independent patient-specific factors into account when studying in situ biomaterial-driven tissue engineering.


Functional organization of the ventral lateral geniculate complex of the tree shrew (Tupaia belangeri): II. Connections with the cortex, thalamus, and brainstem.
Connections of the ventral lateral geniculate complex (GLv) in the tree shrew were traced by anterograde and retrograde transport of WGA-HRP. The results buttress earlier findings that GLv in this species is composed of two main divisions, lateral and medial, each of which differs in its connections with the brainstem and cerebral cortex. The connections of the lateral division (GLv) suggest that it participates in visuosensory functions: it receives input from the retina, striate cortex, pretectum, and retino-recipient layers of the superior colliculus. These connections help clarify the identification of the internal and external subdivisions of GLv inasmuch as projections from both the superior colliculus and pretectum terminate in the external subdivision and each, in turn, receives a projection from the internal subdivision. Connections of the medial division suggest that this part of the nucleus is involved with visuomotor functions. Thus, the medio-caudal subdivision projects to the pontine nuclei, the prerubral field and the central lateral nucleus. The medio-caudal subdivision also receives projections from the lateral cerebellar nucleus, so that the GLv-ponto-cerebello-GLv loop involves mainly one subdivision of GLv. The medio-rostral subdivision receives projections from the pretectum and parietal cortex. Its output is directed primarily at the intermediate and deep layers of the superior colliculus. All of these targets of GLv, the pons, prerubral field, and deep layers of the superior colliculus, are known to play a role in the coordination of head and eye movements. Additional connections of GLv with the vestibular nuclei, intralaminar nuclei, hypothalamus, and facial motor nucleus are also described.


Toll-like receptor 4 modulates myocardial ischaemia-reperfusion injury: Role of matrix metalloproteinases.
Toll-like receptor 4 (TLR4) mediates innate immune responses following endotoxemia and myocardial ischaemia-reperfusion (I/R) injury. Pre-treatment with the major TLR4 ligand lipopolysaccharide (LPS) reduces infarct size. Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) play a crucial role in endotoxemia possibly also determining I/R injury. We investigated the influence of TLR4 on infarct size and assessed the influence of MMP and TIMP regulation on I/R injury. Left anterior descending artery (LAD) occlusion was performed on wild-type (C3H/HeN) and TLR4-deficient (C3H/HeJ) mice. Animals were stimulated with LPS (1 mg/kg) or PBS 16 h ahead of 60 min LAD ligation. After 24 h of reperfusion, triphenyltetrazolium chloride staining was performed and infarct size was measured by planimetry. MMP- and TIMP-mRNA expression were determined by RPA after 3 h of reperfusion. MMP zymographic activity was monitored 6 h after occlusion. TLR4-deficient mice and LPS-treated wild-type mice showed significantly reduced infarct areas. LPS-stimulation significantly increased the overall MMP/TIMP mRNA expression ratio due to elevated MMP-3, -8, -9, and TIMP-1 in wild-type mice. I/R overall reduced the MMP/TIMP ratio due to increased MMP-1, TIMP-1, and -3 mRNA expression. LPS pre-treatment and TLR4-deficiency led to a decreased infarct size. However, infarct area and MMP/TIMP ratio were not correlated. This means that in TLR4-deficient mice MMP/TIMP ratios are not determining the infarct size.


Ion homeostasis and the functional roles of SERCA reactions in stimulus-secretion coupling of the pancreatic beta-cell: A mathematical simulation.
The present paper concerns with ion homeostatic reactions in view of stimulus-secretion coupling of the beta-cell, including Ca2+ fluxes of the endoplasmatic reticulum (ER). A steady state of cytosolic sodium and potassium ion concentrations ([Na+]c and [K+]c, respectively), and of the membrane potential (Delta c phi) can be attained only, if the flux through the electrogenic Na-K pump (JNaK) is balanced electrically, and if JNaK is rather high (about 25% of total ATP consumption at 10 mM glucose). Metabolically caused changes of cellular pH are unlikely, because, on the one hand, CO2 can rapidly leave the cell through cellular membranes, and because ATP cycling cannot produce nor consume protons. A slight decrease of pHc during cellular activity is caused mainly by an increased Ca-H exchange flux through the plasma membrane Ca2+ pump (J PMCA), which might be overcome, however, by H+ transport into secretory granules. The present simulations show that the conductance of ATP-sensitive K+ channels (K ATP) is highly susceptible to changes of [Mg2+]c. As a physical link between the Ca2+ filling state of the ER and the initiation of a depolarising, Ca2+ release-activated current (I CRAN), a metabolite (inositol 1,4,-diphosphate (IP2)) of the inositol 1,4,5-triphosphate (IP3) cycle is introduced. Sufficient ATP for insulin secretion is made available during glucose activation by [IP2] inhibition of a parallel [ATP]c consuming flux through protein biosynthesis (J Pbs). This leads to fast oscillations with a triphasic patterns of [Ca2+]c oscillations. Slow oscillations are initiated by including a Ca2+ leak current through highly uncoupled SERCA3 pumps. Both types of oscillations may superimpose yielding compound bursting and mixed oscillations of [Ca2+]c.


Neuroprotective effect of Convolvulus pluricaulis Choisy in oxidative stress model of cerebral ischemia reperfusion injury and assessment of MAP2 in rats.
Convolvulus pluricaulis Choisy commonly known as Shankhapushpi, is traditionally prescribed for nerve debility, loss of memory, epilepsy and as nervine tonic. Plant also proved to have diverse pharmacological activity but the neuroprotection in ischemic stroke were not found. To investigate the effect of Convolvulus pluricaulis against bilateral common carotid artery (BCCA) occlusion induced cerebral ischemic reperfusion injury. The neuroprotective activity of Convolvulus pluricaulis against bilateral common carotid artery (BCCA) occlusion induced cerebral ischemic reperfusion (I/S) injury. Sprague-Dawley rats of either sex (200-250 g) were divided into nine groups of 8 rats each. Sham and control group, saline treated 10 ml/kg orally. Third group treated with Quercetin 25 mg/kg orally and fourth to ninth groups treated with chloroform and ethanol extract of Convolvulus pluricaulis 100, 200, and 400 mg/kg (p.o.) respectively. Control, Quercetin and extract treated groups underwent 30 min BCCA occlusion and 24 h reperfusion on 10th day but sham underwent same surgery without BCCA occlusion and 24 h reperfusion on 10th day. The antioxidant enzymatic and non-enzymatic levels were estimated by UV spectroscopic method and cerebral infarction area, Blood brain barrier disruption, microtubule-associated protein 2 immunohistochemical and histopathological studies were carried out. The results of the study indicate that the chloroform and ethanol extract of Convolvulus pluricaulis showed neuroprotective activity by a significant decrease in lipid peroxidation (p < 0.001) and an increase in superoxide dismutase (p < 0.01, p < 0.001), catalase (p < 0.01, p < 0.001), glutathione (p < 0.001), and total thiol (p < 0.001) levels in extract-treated groups as compared to control group. Measurement of cerebral infarction area, blood brain barrier disruption, microtubule-associated protein 2 immunohistochemical and histopathological studies further supported the protective effect of the extract. Present study revile that Convolvulus pluricaulis has potent neuroprotection against bilateral common carotid artery (BCCA) occlusion induced cerebral ischemic reperfusion injury.


Multiple colorectal neoplasms in X-linked agammaglobulinemia.
X-linked agammaglobulinemia (XLA) is a primary immunodeficiency disorder caused by germline mutation of the Bruton tyrosine kinase (BTK) gene. It is characterized by disturbed B-cell development, decreased immunoglobulin levels, and increased patient susceptibility to infection. An increased risk of cancer has been suggested, but most reports were described before the identification of BTK gene mutation as the cause of XLA. Here we describe 2 patients with genetically ascertained XLA and multiple colorectal neoplasms, supporting an increased risk of colorectal cancer in XLA and highlighting the potential importance of colorectal surveillance in these patients.


Controllable growth of one-dimensional chiral nanostructures from an achiral molecule.
One achiral molecule of thioxanthone-anthracene (TX-A) is able to self-assemble into chiral nanowires by tuning the solvent with different polarity. Left- and right-handed nanowires with helix structure can be produced from chloroform and acetic ester solution of TX-A respectively. The SEM and TEM images confirm the formation of the helical nanowires. The results indicate that the solvent polarity has great influence in controlling of the self-assembly of organic molecules. The growth process demonstrates a promising pathway to investigate the self-assembling from achiral organic molecules to chiral aggregations.


Voltage noise observed in rods of the turtle retina.
1. Intracellular voltage was recorded from rods in isolated retinae of the snapping turtle, Chelydra serpentina. The voltage was observed during darkness or during uniform illumination of a large retinal area. During darkness the voltage fluctuated continuously about a mean level. The spontaneous fluctuation is termed ;noise'. During illumination the amplitude of the noise was reduced.2. The noise observed during darkness could also be reduced by injecting a hyperpolarizing current into the impaled rod. The noise could be increased by a depolarizing current. The component of the noise that could be altered by polarizing the rod is termed ;voltage-sensitive noise'.3. When voltage-sensitive noise was first minimized by a continuous hyperpolarizing current, bright light produced an additional decrease in the noise. The component of the noise that was eliminated by light, but not eliminated by the injection of current, is termed ;light-sensitive noise'.4. The power density spectrum of voltage-sensitive noise, G(v)(f), could be described by an equation of the form [Formula: see text] tau(M) was approximately 7 msec, which is in good agreement with an apparent membrane time constant of 5-8 msec. The largest value of alpha(v) was 2.1 x 10(-9) V(2) sec.5. The power density spectrum of light-sensitive noise could be described by an equation of the form [Formula: see text] tau(L) was approximately 200-300 msec. The largest value of alpha(L) was 8.0 x 10(-9) V(2) sec.6. The potential maintained during darkness could be altered by superfusing a retina with artificial media of different compositions. Depolarizing the rods by changing the extracellular calcium concentration from 1 to 5 mM increased the voltage-sensitive noise. A similar effect was observed after adding 2 mM lanthanum.7. In contrast, 5 mM cobalt produced a small hyperpolarization and suppressed the voltage-sensitive noise. Injecting a depolarizing current, after exposure to cobalt, re-initiated the voltage-sensitive noise. The ability to elicit voltage-sensitive noise in the presence of cobalt indicates that it was not of synaptic origin.8. The results are consistent with the noise present during dark being produced by two types of channel in the rod membrane. One is controlled by the phototransduction process; each individual channel of this type may be described as having a mean open time of 200-300 msec and a conductance of approx. 6 x 10(-13) Omega(-1). The absorption of one photon closes approx. 100-300 of these channels. The other type of channel is controlled by membrane potential; each individual channel of this type has a mean open time which is less than the membrane time constant of 8 msec.


Reparative strand incision in saponin-permeabilized human fibroblasts.
The damage-directed strand incision step in the nucleotidyl DNA excision-repair pathway (NDERP) was characterized in quiescent monolayer cultures of human fibroblasts in which the plasma membrane was selectively permeabilized with saponin. When permeable normal human fibroblasts (NHF) were incubated in a DNA-repair assay mixture lacking the deoxyribonucleoside triphosphate precursors, the numbers of UV-dependent DNA-strand breaks were increased by about 9-fold consistent with the uncoupling of incision from gap-filling DNA synthesis and ligation. In uncoupled NHF omission of ATP reduced the numbers of UV-dependent strand breaks by 84% confirming the requirement for ATP for reparative strand incision. Time-course experiments indicated that the maximum rate of strand incision occurred in the first 10 min of incubation of permeable cells and diminished to 16-28% of this rate between 30 and 60 min of incubation. The initial rate of incision in permeable NHF was estimated to be 20% of that seen in intact fibroblasts. Dose-response studies indicated an initial saturation of strand incision activity at fluences between 10 and 25 J/m2. In permeable group A xeroderma pigmentosum fibroblasts (XPA) few UV-dependent incisions were produced after 10-25 J/m2. In the xeroderma pigmentosum variant (XPV) strain that we studied, strand incisions saturated at a plateau level that was about twice that seen in the NHF strain suggesting the preservation of a higher level of incision activity after permeabilization. After fluences above 50 J/m2 additional strand incision was observed in all cell strains reflecting the activity of a damage-dependent endodeoxyribonuclease that is independent of the NDERP. Saponin-treated fibroblasts were also permeable to pancreatic deoxyribonuclease I and the UV-DNA endonuclease from M. luteus indicating that these preparations may be used for in vitro complementation.


Biochemical Characterization of VIM-39, a VIM-1-Like Metallo-β-Lactamase Variant from a Multidrug-Resistant Klebsiella pneumoniae Isolate from Greece.
VIM-39, a VIM-1-like metallo-β-lactamase variant (VIM-1 Thr33Ala His224Leu) was identified in a clinical isolate of Klebsiella pneumoniae belonging to sequence type 147. VIM-39 hydrolyzed ampicillin, cephalothin, and imipenem more efficiently than did VIM-1 and VIM-26 (a VIM-1 variant with the His224Leu substitution) because of higher turnover rates.


Validity and reliability assessment of the Compliance with Standard Precautions Scale Arabic version in Saudi nursing students.
Strict compliance with standard precautions (SPs) is warranted to ensure the safety of patients in healthcare facilities. Nursing students (NSs), who are regarded as nurses in training, potentially play a role in cross-contamination in the hospital. NSs are also at high risk of acquiring infections in cases of ineffective compliance with SPs. Thus, an assessment of NSs' compliance with SPs should be conducted on a regular basis, which necessitates a valid and reliable tool. This study was conducted to assess the validity and reliability of the Compliance with Standard Precautions Scale Arabic version (CSPS-A) in Saudi NSs. A convenient sample of 230 respondents (158 NSs and 72 staff nurses) was included in this descriptive, cross-sectional study. The CSPS English version was translated into the Arabic language following a recommended guideline from cross-cultural adaptation and translation instruments. The Cronbach's alpha, the intraclass correlation coefficient (ICC) of the test-retest scores, and the item-total correlations (ITC) were computed to establish the reliability. Content validity and construct validity by the known-groups method and hypothesis testing method were performed. The CSPS-A exhibited good internal consistency and reliability (Cronbach's alpha, 0.89; ICC, 0.88; ITCs, 0.325-0.728). A satisfactory content and construct validity was also reported. The CSPS-A is a valid and reliable tool that can measure the compliance to SPs among NSs in Saudi Arabia.


Establishment and validation of a risk prediction model incorporating concentrations of linezolid and its metabolite PNU142300 for linezolid-induced thrombocytopenia.
Linezolid-induced thrombocytopenia is the main factor restricting the clinical application of linezolid. To investigate the relationship between PNU-14230 concentration and linezolid-induced thrombocytopenia and further develop and validate a risk model for predicting linezolid-induced thrombocytopenia. A regression model was constructed to predict the occurrence of linezolid-induced thrombocytopenia, and further externally validated. The predictive performance was evaluated by receiver operating characteristic curve and Hosmer-Lemeshow test. Linezolid Cmin and PNU-142300 concentrations were compared for different kidney function groups. The Kaplan-Meier method was used to estimate the difference in cumulative incidence of linezolid-induced thrombocytopenia among different kidney function patients. In the derivation (n = 221) and validation (n = 158) cohorts, 28.5% and 24.1% of critically ill patients developed linezolid-induced thrombocytopenia. Logistic regression analysis indicated that the independent risk factors were linezolid Cmin, PNU-142300 concentration, baseline platelet count, renal insufficiency (RI) and continuous venovenous haemofiltration (CVVH). The AUC for the risk model was 0.901, and the model was good (P = 0.633). The model also showed good discrimination (AUC 0.870) and calibration (P = 0.282) in the external validation cohort. Compared with normal kidney function patients, patients with RI and CVVH had higher linezolid Cmin and PNU-142300 concentrations (P < 0.001) and higher cumulative incidence of linezolid-induced thrombocytopenia (P < 0.001). PNU142300 concentration, as well as linezolid Cmin, might identify patients at risk of linezolid-induced thrombocytopenia. The risk prediction model had good predictive performance for linezolid-induced thrombocytopenia development. Concentrations of linezolid and PNU-142300 accumulated in patients with RI and CVVH.


Predictive model to describe the combined effect of pH and NaCl on apparent heat resistance of Bacillus stearothermophilus.
The combined effect of pH and NaCl on the apparent thermal resistance of Bacillus stearothermophilus ATCC 12980 spores was studied. Spores were heated at different temperatures (115-125 degrees C) in mushroom substrate, acidified using glucono-delta-lactone to different pH levels (from 5.75 to 6.7), which contained concentrations of NaCl that ranged from 0.5 to 3% (w/v). The recovery medium was acidified to the same pH level and contained the same NaCl concentration as the heating menstruum. A factorial experimental design allowed a predictive model to be developed, which described the combined effect of heating temperature, pH and NaCl on the thermal resistance of B. stearothermophilus spores. Predictions from the model provided a valid description of the data used to generate the model, and agreed with observations from the literature and from an independent experiment performed using asparagus and bean substrates.


A comparison of the effects of PGI2, PGE2 and PGH2 on the cyclic nucleotide levels in rat anterior pituitary gland in vitro.
Rat anterior pituitary explants were incubated with PGI2, PGH2 and PGE2 in the presence of theophylline (1mM) and the production of cyclic AMP was measured. PGE2 was found to be about 20 times more potent than PGI2 while PGH2 was slightly more effective than PGI2. The results suggest that PGI2 does not play a AMP was measured. PGE was found to be about 20 times more potent than physiological role in cyclic AMP mediated events in the rat anterior pituitary.


On collective self-healing and traces: How can swarm intelligence help us think differently about team adaptation?
Health care teams are increasingly forced to navigate complex challenges to achieve their collective aim of delivering high-quality, safe patient care. The teamwork literature has struggled to develop strategies that promote effective adaptive behaviours among health care teams. In part, this challenge stems from the fact that truly collective adaptive behaviour requires members of the teams to abandon the human urge to act self-sufficiently. Nature contains striking examples of collective behaviour as seen in social insects, fish and bird colonies. This collective behaviour is known as Swarm Intelligence (SI). SI remains poorly described in the health care team literature and its potential benefits hidden. In this cross-cutting edge paper, I explore the principles of SI as they pertain to systemic or collective adaptation in human teams. In particular, I consider the principles of trace-based communication and collective self-healing and what they might offer to team adaptation researchers in medical education. From a SI perspective, a solution to a problem emerges as a result of the collective action of the members of the swarm, not the individual action. This collective action is achieved via four principles: direct and indirect communication, awareness, self-determination and collective self-healing. Among those principles, trace-based communication and collective self-healing have been purposefully used by other industries to foster team adaptation. Trace-based communication relies on leaving 'traces' in the environment to drive the behaviour of others. Collective self-healing is the ability of the swarm to cope with failure and adapt to changes by permitting swarm members to be interchangeable. While allowing teams to rely on indirect communication and to be interchangeable might create discomfort to our ways of thinking, teams outside health care are demonstrating their value to advance human teamwork. SI offers a helpful analogy and a constructive language for thinking about team adaptation.


Chromosomal studies in four species of genus Chaunus (Bufonidae, Anura): localization of telomeric and ribosomal sequences after fluorescence in situ hybridization (FISH).
Fluorescence in situ hybridization (FISH) using telomeric and ribosomal sequences was performed in four species of toad genus Chaunus: C. ictericus, C. jimi, C. rubescens and C. schneideri. Analyses based on conventional, C-banding and Ag-NOR staining were also carried out. The four species present a 2n = 22 karyotype, composed by metacentric and submetacentric chromosomes, which were indistinguishable either after conventional staining or banding techniques. Constitutive heterochromatin was predominantly located at pericentromeric regions, and telomeric sequences (TTAGGG)(n) were restricted to the end of all chromosomes. Silver staining revealed Ag-NORs located at the short arm of pair 7, and heteromorphism in size of NOR signals was also observed. By contrast, FISH with ribosomal probes clearly demonstrated absence of any heteromorphism in size of rDNA sequences, suggesting that the difference observed after Ag-staining should be attributed to differences in chromosomal condensation and/or gene activity rather than to the number of ribosomal cistrons.


Paediatric cardiology programs in countries with limited resources: how to bridge the gap.
Establishing paediatric cardiology service in a country with limited resources like Sudan is a challenging task. A paediatric cardiac team was formed then the services in different disciplines were gradually established. Echocardiography (echo) clinics were founded in tertiary and peripheral hospitals. Cardiac catheterization (cath) was established at the Sudan Heart Centre (SHC) in 2004 and over 400 procedures had been performed including interventional catheterization like pulmonary valve dilatation, patent ductus arteriosus and atrial septal defect device closure. Congenital heart surgery started in 2001, currently 200 cases are done each year including closed procedures as well as open heart procedures for patients weighing more than 8 kg. Cardiology-cardiac surgery as well as adult congenital heart disease meetings were held and contributed positively to the services. The cardiology-cardiac surgery scientific club meeting was founded as a forum for academic discussions. A fellowship program was established in 2004 and included seven candidates trained in paediatric cardiology and intensive care. Two training courses had been established: congenital heart disease echo and paediatric electrocardiogram interpretation. Links with regional and international cardiac centres had important roles in consolidating our program. Significant obstacles face our service due to the small number of trained personnel, high cost of procedures, the lack of regular supplies and lack of cardiac intensive care facilities for young infants. Bridging the huge gap needs extensive official as well as non-governmental efforts, training more staff, supporting families and collaboration with regional and international centres.


Transcriptome profiling reveals that feeding wild zooplankton to larval Atlantic cod (Gadus morhua) influences suites of genes involved in oxidation-reduction, mitosis, and selenium homeostasis.
Larval nutrition and growth are key issues for wild and cultured cod. While it was shown previously that larval cod fed wild zooplankton grow faster than those fed only rotifers, the mechanisms involved in this enhanced growth are not completely understood. We used microarrays to identify larval cod transcripts that respond to feeding with small amounts of wild zooplankton (5-10 % of live prey items). The larval transcriptome was compared between 3 treatment groups [fed rotifers (RA), rotifers with protein hydrolysate (RA-PH), or rotifers with zooplankton (RA-Zoo)] at 9-10 mm length [26-30 days post-hatch (dph)] to identify a robust suite of zooplankton-responsive genes (i.e. differentially expressed between RA-Zoo and both other groups). The microarray experiment identified 147 significantly up-regulated and 156 significantly down-regulated features in RA-Zoo compared with both RA and RA-PH. Gene ontology terms overrepresented in the RA-Zoo responsive gene set included "response to selenium ion" and several related to cell division and oxidation-reduction. Ten selenoprotein-encoding genes, and 2 genes involved in thyroid hormone generation, were up-regulated in RA-Zoo compared with both other groups. Hierarchical clustering of RA-Zoo responsive genes involved in oxidation-reduction and selenium homeostasis demonstrated that only the zooplankton treatment had a considerable and consistent impact on the expression of these genes. Fourteen microarray-identified genes were selected for QPCR involving 9-13 mm larvae, and 13 of these were validated as differentially expressed between RA-Zoo and both other groups at ~9 mm. In contrast, in age-matched (34-35 dph; ~11 mm RA and RA-PH, ~13 mm RA-Zoo) and size-matched (~13 mm) older larvae, only 2 and 3 genes, respectively, showed the same direction of RA-Zoo-responsive change as in ~9 mm larvae. The modulation of genes involved in selenium binding, redox homeostasis, and thyroid hormone generation in ~9 mm RA-Zoo larvae in this study may be in response to the relatively high levels of selenium, iodine, and LC-PUFA (potentially causing oxidative stress) in zooplankton. Nonetheless, only a subset of zooplankton-responsive genes in ~9 mm larvae remained so in older larvae, suggesting that the observed transcriptome changes are largely involved in initiating the period of growth enhancement.


Nox2 and Cyclosporine-Induced Renal Hypoxia.
We hypothesized that nicotinamide adenosine diphosphate oxidase 2 (Nox2) plays an important role in cyclosporine A (CsA)-induced chronic hypoxia. We tested this hypothesis in Fisher 344 rats, C57BL/6 J wild type and Nox2-/- mice, and in liver transplant recipients with chronic CsA nephrotoxicity. We used noninvasive molecular imaging (blood oxygen level-dependent magnetic resonance imaging and dynamic contrast-enhanced magnetic resonance imaging) and molecular diagnostic tools to assess intrarenal oxygenation and perfusion, and the molecular phenotype of CsA nephrotoxicity. We observed that chemical and genetic inhibition of Nox2 in rats and mice resulted in the prevention of CsA-induced hypoxia independent of regional perfusion (blood oxygen level-dependent magnetic resonance imaging and dynamic contrast-enhanced magnetic resonance imaging, pimonidazole, HIF-1α). Nicotinamide adenosine diphosphate oxidase 2 knockout was also associated with decreased oxidative stress (Nox2, HIF-1α, hydrogen peroxide, hydroxynonenal), and fibrogenesis (α-smooth muscle actin, picrosirius red, trichrome, vimentin). The molecular signature of chronic CsA nephrotoxicity using transcriptomic analyses demonstrated significant changes in 40 genes involved in injury repair, metabolism, and oxidative stress in Nox2-/- mice. Immunohistochemical analyses of kidney biopsies from liver transplant recipients with chronic CsA nephrotoxicity showed significantly greater Nox2, α-smooth muscle actin and picrosirius levels compared with controls. These studies suggest that Nox2 is a modulator of CsA-induced hypoxia upstream of HIF-1α and define the molecular characteristics that could be used for the diagnosis and monitoring of chronic calcineurin inhibitor nephrotoxicity.


A supramolecular photosensitizer derived from an Arene-Ru(II) complex self-assembly for NIR activated photodynamic and photothermal therapy.
Effective photosensitizers are of particular importance for the widespread clinical utilization of phototherapy. However, conventional photosensitizers are usually plagued by short-wavelength absorption, inadequate photostability, low reactive oxygen species (ROS) quantum yields, and aggregation-caused ROS quenching. Here, we report a near-infrared (NIR)-supramolecular photosensitizer (RuDA) via self-assembly of an organometallic Ru(II)-arene complex in aqueous solution. RuDA can generate singlet oxygen (1O2) only in aggregate state, showing distinct aggregation-induced 1O2 generation behavior due to the greatly increased singlet-triplet intersystem crossing process. Upon 808 nm laser irradiation, RuDA with excellent photostability displays efficient 1O2 and heat generation in a 1O2 quantum yield of 16.4% (FDA-approved indocyanine green: ΦΔ = 0.2%) together with high photothermal conversion efficiency of 24.2% (commercial gold nanorods: 21.0%, gold nanoshells: 13.0%). In addition, RuDA-NPs with good biocompatibility can be preferably accumulated at tumor sites, inducing significant tumor regression with a 95.2% tumor volume reduction in vivo during photodynamic therapy. This aggregation enhanced photodynamic therapy provides a strategy for the design of photosensitizers with promising photophysical and photochemical characteristics.