Lead-sulfur interaction induced damp and water stability in pure formamidinium lead triiodide.
Research efforts in various multitudes have been demonstrated to stabilize methylammonium (MA)- and bromide (Br)-free formamidinium lead triiodide (FAPI) perovskite thin films. Despite these commendable efforts, pure FAPI perovskite thin film is prone to critical phase-transition issues due to its thermodynamically stable non-perovskite phase (2H). Here, in this work, we propose a rational additivization strategy to overcome this challenge. Our multifunctional ammonium salt containing a sulfur heteroatom shifts the thermodynamic stability from the 2H phase to an intermediate phase closer to the cubic phase. Along with the high crystallinity, micron-sized grains with preferred (00h) facet orientation stem the Pb…S interaction to offer exceptional stability against high relative humidity, direct water incursion, and shelf-life aging. Our findings through experimental and theoretical studies substantiate the role of Pb…S interaction in stabilizing the perovskite cubic phase and the stoichiometric distribution of elemental components.


L-Asparaginase of Leishmania donovani: Metabolic target and its role in Amphotericin B resistance.
Emergence of Amphotericin B (AmB) resistant Leishmania donovani has posed major therapeutic challenge against the parasite. Consequently, combination therapy aimed at multiple molecular targets, based on proteome wise network analysis has been recommended. In this regard we had earlier identified and proposed L-asparaginase of Leishmania donovani (LdAI) as a crucial metabolic target. Here we report that both LdAI overexpressing axenic amastigote and promastigote forms of L. donovani survives better when challenged with AmB as compared to wild type strain. Conversely, qRT-PCR analysis showed an upregulation of LdAI in both forms upon AmB treatment. Our data demonstrates the importance of LdAI in imparting immediate protective response to the parasite upon AmB treatment. In the absence of structural and functional information, we modeled LdAI and validated its solution structure through small angle X-ray scattering (SAXS) analysis. We identified its specific inhibitors through ligand and structure-based approach and characterized their effects on enzymatic properties (Km, Vmax, Kcat) of LdAI. We show that in presence of two of the inhibitors L1 and L2, the survival of L. donovani is compromised whereas overexpression of LdAI in these cells restores viability. Taken together, our results conclusively prove that LdAI is a crucial metabolic enzyme conferring early counter measure against AmB treatment by Leishmania.


Chlorophyll a formation in the chlorophyll b reductase reaction requires reduced ferredoxin.
The reduction of chlorophyllide b and its analogue zinc pheophorbide b in etioplasts of barley (Hordeum vulgare L.) was investigated in detail. In intact etioplasts, the reduction proceeds to chlorophyllide a and zinc pheophorbide a or, if incubated together with phytyldiphosphate, to chlorophyll a and zinc pheophytin a, respectively. In lysed etioplasts supplied with NADPH, the reduction stops at the intermediate step of 7(1)-OH-chlorophyll(ide) and Zn-7(1)-OH-pheophorbide or Zn-7(1)-OH-pheophytin. However, the final reduction is achieved when reduced ferredoxin is added to the lysed etioplasts, suggesting that ferredoxin is the natural cofactor for reduction of chlorophyll b to chlorophyll a. The reduction to chlorophyll a requires ATP in intact etioplasts but not in lysed etioplasts when reduced ferredoxin is supplied. The role of ATP and the significance of two cofactors for the two steps of reduction are discussed.


Influences of nifedipine on vascular conductance of portally drained viscera, liver and kidney.
Nifedipine is a potent vasodilator, which induces various dose-related hemodynamic responses in the gastrointestinal tract, liver and kidney. Blood flow was assessed in anesthetized dogs with non-cannulating electromagnetic flow sensors. Nifedipine was injected into a brachial vein at intervals of 2 min in graded doses from 1-16 micrograms.kg-1. The pulsatile flow and pressure signals were analog-digital converted and processed by a computer program to obtain mean flow (ml.min-1) and mean pressure (mm Hg) values for every 3 s. The vascular data are presented as conductance. The arterial pressure decreased 45% while vascular conductance in vessels of the stomach, duodenum, small and large bowel increased between 75 and 128%. No response was observed in the splenic and common hepatic artery. An increase of resistance in the hepatic artery proper, while portal flow increased suggests that there were changes in liver metabolism. Although the literature indicates a decrease in renal vascular resistance, we encountered a decrease in flow but no change in resistance. Neither did we find an increase in heart rate. We propose that small intravenous doses of nifedipine reduce the afterload of the left ventricle.


Disruption of the proximal anastomosis of axillobifemoral grafts: two case reports.
The use of axillobifemoral grafting in the treatment of infected aortic prostheses or in the treatment of patients at high risk for aortoiliac occlusive disease has become a widely accepted treatment modality. Few complications involving the proximal anastomosis have been reported. The present report describes two patients who suffered proximal anastomotic dehiscence several weeks after surgery when they fully extended and raised the affected arm.


Early phenotypic features of aniridia-associated keratopathy and association with PAX6 coding mutations.
To investigate corneal phenotype in aniridia-associated keratopathy (AAK) including its earliest manifestations, in relation to PAX6 mutational status. 46 subjects (92 eyes) with congenital aniridia from a German registry were examined using slit lamp biomicroscopy, anterior segment optical coherence tomography, contact esthesiometry and in vivo confocal microscopy. Cytogenetic analysis was conducted by Sanger sequencing of PAX6 exons and/or MLPA analysis. Measured parameters included AAK grade, distance-corrected visual acuity (DCVA), central corneal thickness (CCT), corneal sensitivity, subbasal nerve density, mature dendritic cell (DC) density and corneal epithelial phenotype. 46 subjects (age range: 1-64 years) were examined, including 23 (50%) children under the age of 18. Five subjects (11.1%) with absent PAX6 coding mutation (non-PAX6 cases) had mild AAK (Grade 0-1) into the fourth decade of life and maintained corneal epithelial phenotype, greater subbasal nerve density (16.8 mm/mm2 vs. 3.58 mm/mm2, P = 0.01) and better corneal sensitivity (41 ± 11 mm vs. 28 ± 12 mm, P = 0.03) relative to those with PAX6 coding mutations. In five subjects, corneal endothelial cell density ranged from 3245 to 4399 cells/mm2. Independent of mutational status, an increased CCT, over tenfold increased mature DC density and reduced corneal sensitivity characterized all subjects. PAX6 coding mutations influence AAK phenotype and progression from the earliest stages of life. A minimal keratopathy present in 100% of congenital aniridia cases is independent of the specific mutation and consists of increased corneal thickness, reduced touch sensitivity, and increased ocular surface immune activity.


Receptor for advanced glycation end products (RAGE) in vascular and inflammatory diseases.
Historically, the receptor for advanced glycation end products (RAGE) was thought to exclusively play an important role under hyperglycemic conditions. However, more and more evidence suggests that RAGE in fact is an inflammation perpetuating multi-ligand receptor and participates actively in various vascular and inflammatory diseases even in normoglycaemic conditions. Various ligands include advanced glycation end products (AGEs), S100 proteins and amphoterins etc. Besides full-length RAGE, numerous truncated forms of the receptor have also been described including the well-characterized soluble RAGE (sRAGE). sRAGE has an ability to act as a decoy to avoid interaction of RAGE with its pro-inflammatory ligands. Ligand engagement of RAGE activates multiple signaling pathways and also forms a positive feedback loop for its own enhanced expression. This review will discuss the role of multi-ligand receptor i.e. RAGE in context to various vascular diseases, which have a pathophysiologically important inflammatory component in normoglycaemic conditions.


Predicting Changes in Systolic and Diastolic Blood Pressure of Hypertensive Patients in Indonesia Using Machine Learning.
This retrospective study investigated factors that influence the occurrence of decreased systolic and diastolic blood pressure including sociodemographic and economic factors, hypertension duration, cigarette consumption, alcohol consumption, duration of smoking, type of cigarettes, exercise, salt consumption, sleeping pills consumption, insomnia, and diabetes. These factors were applied to predict the reality of systolic and diastolic decrease using the machine learning algorithm Naïve Bayes, artificial neural network, logistic regression, and decision tree. The increase in blood pressure, both systolic and diastolic, is very harmful to the health because uncontrolled high systolic and diastolic blood pressure can cause various diseases such as congestive heart failure, kidney failure, and cardiovascular disease. There have been many studies examining the factors that influence the occurrence of hypertension, but few studies have used machine learning to predict hypertension. The machine learning models performed well and can be used for predicting whether a person with hypertension with certain characteristics will experience a decrease in their systolic or diastolic blood pressure after treatment with antihypertensive drugs.


Are quantitative trait-dependent sampling designs cost-effective for analysis of rare and common variants?
Use of trait-dependent sampling designs in whole-genome association studies of sequence data can reduce total sequencing costs with modest losses of statistical efficiency. In a quantitative trait (QT) analysis of data from the Genetic Analysis Workshop 17 mini-exome for unrelated individuals in the Asian subpopulation, we investigate alternative designs that sequence only 50% of the entire cohort. In addition to a simple random sampling design, we consider extreme-phenotype designs that are of increasing interest in genetic association analysis of QTs, especially in studies concerned with the detection of rare genetic variants. We also evaluate a novel sampling design in which all individuals have a nonzero probability of being selected into the sample but in which individuals with extreme phenotypes have a proportionately larger probability. We take differential sampling of individuals with informative trait values into account by inverse probability weighting using standard survey methods which thus generalizes to the source population. In replicate 1 data, we applied the designs in association analysis of Q1 with both rare and common variants in the FLT1 gene, based on knowledge of the generating model. Using all 200 replicate data sets, we similarly analyzed Q1 and Q4 (which is known to be free of association with FLT1) to evaluate relative efficiency, type I error, and power. Simulation study results suggest that the QT-dependent selection designs generally yield greater than 50% relative efficiency compared to using the entire cohort, implying cost-effectiveness of 50% sample selection and worthwhile reduction of sequencing costs.


Late preterm infants, early term infants, and timing of elective deliveries.
Delivery of infants who are physiologically mature and capable of successful transition to the extrauterine environment is an important priority for obstetric practitioner. A corollary of this goal is to avoid iatrogenic complications of prematurity and maternal complications from delivery. The purpose of this review is to describe the consequences of birth before physiologic maturity in late preterm and term infants, to identify factors contributing to the decline in gestational age of deliveries in the United States, and to describe strategies to reduce premature delivery of late preterm and early term infants.


Specific immune response to Parietaria judaica plant profilin: a low T cell proliferative response supports high IgE and skin prick test.
allergic disease caused by Parietaria judaica (Pj) has been widely documented in Mediterranean area. Profilins have been identified as widely distributed allergenic proteins. The role of Pj profilin in specific immune response in Pj-sensitized patients is unknown. skin prick test and determination of specific and total IgE levels in serum were performed in all patients (n = 28) and non-allergic controls (n = 18). Peripheral blood mononuclear cells (PBMC) were isolated from both groups and stimulated with crude extract or highly purified Pj profilin. The production of type I and type II cytokines was determined by specific and polyclonal stimuli in patients and controls. T-cell lines specific to Pj profilin were established and cross-reactivity with another highly purified profilin from Phleum pratense (Phl p) was evaluated. Pj profilin-sensitized patients showed a small but significantly increased in T-cell proliferative response to this profilin compared with non-atopic controls. The production of interleukin (IL)-4 and interferon (IFN)-γ in response to the specific stimulus was undetectable. However, the production of IL-4 in response to a polyclonal stimulus [phytohemagglutinin (PHA)] was significantly higher in atopic patients than in controls. The T-cell response did not correlate with the magnitude of response to skin prick tests with Pj profilin or with Pj-specific serum IgE levels. In addition, the production of IL-4 in response to a polyclonal stimulus (PHA) did not correlate with the individual skin prick tests to Pj profilin or with Pj-specific IgE levels in serum. The T-cell lines tested showed no cross-reactivity with Phl p profilin. our results suggest that Pj profilin is partly responsible for the T-cell-mediated response in patients allergic to Pj. The high skin reactivity to Pj profilin is these patients was accompanied by a small increase in the T-cell response to this profilin. The response was highly specific since Pj profilin specific T-cell lines showed no cross-reactivity with a highly homologous profilin from Phl p. The lack of correlation between the proliferative T-cell response and polyclonal IL-4 production with allergen-specific serum IgE and skin reactivity probably indicates that some of the responding T-cells may be involved in immune reactions other than those supporting IgE production.


A nonsense mutation (G15059A) in the cytochrome b gene in a patient with exercise intolerance and myoglobinuria.
We describe a new mitochondrial DNA mutation in the cytochrome b gene in a patient presenting with progressive exercise intolerance and myoglobinuria associated with complex III deficiency in muscle. The point mutation results in the replacement of a glycine at amino acid position 190 with a stop codon. This change predicts premature termination of translation, leading to a truncated protein missing 244 amino acids at the C-terminus of cytochrome b. The mutation fulfills all the accepted criteria for pathogenicity, suggesting that this is the primary cause of the myopathy in the patient.


Disseminated disease severity as a measure of virulence of Mycobacterium tuberculosis in the guinea pig model.
Virulence is the measure of pathogenicity of a microorganism as determined by its ability to invade host tissues and to produce severe disease. In the low-dose aerosol guinea pig model the virulence of multiple strains of Mycobacterium tuberculosis was determined by measuring time of survival, bacterial loads in target organs, and the severity of pulmonary and extra-pulmonary lesions. Erdman K01, CSU93/CDC1551 and HN878 had shorter survival times compared to the common laboratory strain H37Rv. After 30 days of the infection bacilli had disseminated from the lungs resulting in microscopically visible lesions in peribronchial lymph nodes, peripancreatic lymph nodes, spleen, liver, pancreas, adrenal and heart. The extent of the lesion necrosis paralleled virulence when survival times were used as a measure as Erdman K01 and the two clinical isolates caused more necrosis and resulted in sooner death in infected animals than the H37Rv. The extent of extra-pulmonary lesion necrosis was a better predictor of virulence than the number of viable bacilli in the tissue. Overall, this study emphasizes the point that extra-pulmonary disease is a prominent feature of the guinea pig model and dissemination to organs not normally assayed such as the heart and adrenal glands should be taken into account in the assessment of the disease process.


Orientation and its consequences for Copiapoa (Cactaceae) in the Atacama Desert.
Three species of the barrel cactus Copiapoa (C. cinerea, C. columna-alba, C. haseltoniana) were investigated in their native habitats along the cool, arid coastal regions of the Atacama Desert in northern Chile. All species orient towards the north with a high degree of precision. Two consequences of adaptive value result from this northerly orientation. First, tissue temperatures of the meristematic and floral regions on the tip of the cactus receive high solar radiation loads which result in high temperatures (30°-40°C) relative to air temperatures (15°-20°) during winter and spring months when adequate soil moisture for growth is available. Second, absorption of solar radiation by the sides of the cactus is minimized, which reduces both the potential detrimental effects of light and heat load on the cactus and probably balances daily quanta absorbed for photosynthesis with nighttime CO2 uptake rates during drought stress periods.


The Natural Course of Cigarette Smoking among Adolescent Daily Smokers in France and Quebec.
To describe the natural course of cigarette smoking and nicotine dependence (ND) over 1-year in daily smokers ages 15 to 17 living in different social contexts. Cigarette smoking and ND indicators were measured at baseline and 3- and 12-months thereafter among 95 daily smokers with a total of 123 observations from the Nicotine Dependence in Teens (NDIT) Study in Quebec, Canada, and in 111 daily smokers from the TABagisme chez les ADOlescents (TABADO) Study in Lorraine, France. NDIT and TABADO participants initiated smoking a mean (SD) of 3.9 (1.6) and 3.7 (2.0) years prior to baseline, respectively. Despite baseline differences in age, sex, age at initiation, number of cigarettes smoked per day and social context, 85% of participants in both samples reported cravings and responded "yes" to "felt like you really need a cigarette". Mean (SD) number of cigarettes smoked per day increased from 9.4 (8.1) to 11.8 (8.0) over 1 year in NDIT (adjusted mean difference (95% CI) = 2.4 (0.8, 3.0)), and from 11.5 (6.5) to 13.5 (6.7) in TABADO (adjusted mean difference (95% CI) = 2.0 (0.8, 3.1)). However, most ND indicators in both samples were stable over time and cessation was infrequent. Despite notable differences across samples, the natural course of cigarettes smoked per day, ND symptoms and cessation was similar, suggestive of an underlying biologic rather than social process. To quit, adolescents who smoke daily will likely need (pharmacologic) intervention to counter the biological mechanisms underpinning ND, as well as complementary strategies targeting the social context such as creating social environments favoring cessation success.


Readiness of health facilities to manage individuals infected with COVID-19, Uganda, June 2021.
The COVID-19 pandemic overwhelmed the capacity of health facilities globally, emphasizing the need for readiness to respond to rapid increases in cases. The first wave of COVID-19 in Uganda peaked in late 2020 and demonstrated challenges with facility readiness to manage cases. The second wave began in May 2021. In June 2021, we assessed the readiness of health facilities in Uganda to manage the second wave of COVID-19. Referral hospitals managed severe COVID-19 patients, while lower-level health facilities screened, isolated, and managed mild cases. We assessed 17 of 20 referral hospitals in Uganda and 71 of 3,107 lower-level health facilities, selected using multistage sampling. We interviewed health facility heads in person about case management, coordination and communication and reporting, and preparation for the surge of COVID-19 during first and the start of the second waves of COVID-19, inspected COVID-19 treatment units (CTUs) and other service delivery points. We used an observational checklist to evaluate capacity in infection prevention, medicines, personal protective equipment (PPE), and CTU surge capacity. We used the "ReadyScore" criteria to classify readiness levels as > 80% ('ready'), 40-80% ('work to do'), and < 40% ('not ready') and tailored the assessments to the health facility level. Scores for the lower-level health facilities were weighted to approximate representativeness for their health facility type in Uganda. The median (interquartile range (IQR)) readiness scores were: 39% (IQR: 30, 51%) for all health facilities, 63% (IQR: 56, 75%) for referral hospitals, and 32% (IQR: 24, 37%) for lower-level facilities. Of 17 referral facilities, two (12%) were 'ready' and 15 (88%) were in the "work to do" category. Fourteen (82%) had an inadequate supply of medicines, 12 (71%) lacked adequate supply of oxygen, and 11 (65%) lacked space to expand their CTU. Fifty-five (77%) lower-level health facilities were "not ready," and 16 (23%) were in the "work to do" category. Seventy (99%) lower-level health facilities lacked medicines, 65 (92%) lacked PPE, and 53 (73%) lacked an emergency plan for COVID-19. Few health facilities were ready to manage the second wave of COVID-19 in Uganda during June 2021. Significant gaps existed for essential medicines, PPE, oxygen, and space to expand CTUs. The Uganda Ministry of Health utilized our findings to set up additional COVID-19 wards in hospitals and deliver medicines and PPE to referral hospitals. Adequate readiness for future waves of COVID-19 requires additional support and action in Uganda.


Interaction of psychoactive drugs with the Mg+2 requiring ATPase associated with isolated synaptic vesicles.
A large series of CNS active drugs have been tested for their possible effect on the Mg+2 dependent ATPase associated with isolated synaptic vesicles. Antipsychotic agents and cocaine showed inhibition, of a non-competitive type. The implications of this finding for a synaptic theory of psychosis are discussed.


Dynamic cellular changes in acute kidney injury caused by different ischemia time.
Ischemia reperfusion injury (IRI), often related to surgical procedures, is one of the important causes of acute kidney injury (AKI). To decipher the dynamic process of AKI caused by IRI (with prolonged ischemia phase), we performed single-cell RNA sequencing (scRNA-seq) of clinically relevant IRI murine model with different ischemic intervals. We discovered that Slc5a2hi proximal tubular cells were susceptible to AKI and highly expressed neutral amino acid transporter gene Slc6a19, which was dramatically decreased over the time course. With the usage of mass spectrometry-based metabolomic analysis, we detected that the level of neutral amino acid isoleucine dropped off in AKI mouse plasma metabolites. And the reduction of plasma isoleucine was also verified in patients with cardiac surgery-associated acute kidney injury (CSA-AKI). The findings advanced the understanding of dynamic process of AKI and introduced reduction of isoleucine as a potential biomarker for CSA-AKI.


Differential metabolism of sodium azide in maize callus and germinating embryos.
Sodium azide is a potent mutagen of maize (Zea mays L.) kernels that may have potential as a point mutagen for inducing biochemical mutations in maize tissue cultures. Azide mutagenicity was evaluated in friable, embryogenic maize callus and a nonregenerable maize suspension culture by determining the number of resistant variant cell lines able to grow on media containing inhibitory concentrations of lysine plus threonine (LT). The number of LT-resistant variants selected from either culture type did not increase in response to azide treatment. In addition, there was no increase in somatic mutations in more than 100 plants regenerated from azide treated LT-resistant lines. The levels of mutagenic metabolite of azide (presumably azidoalanine), were determined by bioassay in the two azide-treated maize callus types and compared to levels of mutagenic metabolite in embryos isolated from azide-treated kernels. The two types of maize tissue cultures and isolated embryos contained similar levels of mutagenic metabolite 4 h after azide treatment indicating similar uptake and conversion of azide to mutagenic metabolite in the three tissues. Mutagenic metabolite in azide-treated embryos did not significantly decrease after 40 h. However, mutagenic metabolite levels in both azide-treated tissue cultures decreased to near background levels within 20 h providing evidence for rapid metabolism of the azide mutagenic metabolite. The lack of evidence for azide mutagenicity in maize callus and its known potent mutagenicity in kernels appears to be associated with specific differences in azide metabolism between callus tissues and kernel embryos.


Stem bromelain: amino acid sequence and implications for weak binding of cystatin.
The amino acid sequence of stem bromelain, the major cysteine proteinase from pineapple stem is described. It shows that the enzyme is a member of the papain superfamily of cysteine proteinases, but is not very closely related to any other known member of this group. The sequence shows mutation or deletion of several residues that have been conserved in cysteine proteinases examined previously, including Asn-175 (papain). We suggest that some of these changes have the effect of altering the active-site geometry of stem bromelain, and that this accounts for the resistance of the enzyme to inhibition by cystatins and E-64[L-3-carboxy-2,3-trans-epoxypropionylleucylamido(4-guanidino)b utane].


Water soluble heterometallic potassium-dioxidovanadium(V) complexes as potential antiproliferative agents.
Two water soluble heterometallic potassium–dioxidovanadium polymers, [KVO2(L1)]n (1) and [KVO2(L2)(H2O)]n (2) [H2L1= (2,3-dihydroxybenzylidene)-2-hydroxybenzohydrazide and H2L2=(2,3-dihydroxybenzylidene)benzohydrazide], have been synthesized and characterized by IR, NMR, elemental analysis and single crystal X-ray diffraction. The antiproliferative potentials of 1 and 2 were examined towards human colorectal carcinoma (HCT116), and lung (A549) and breast (MCF7) adenocarcinoma cell lines. 1 exhibits a high cytotoxic activity against colorectal carcinoma cells (HCT116), with IC50 lower than those for cisplatin.


[Management of antiphospholipid syndrome].
The prevention of thrombosis in the antiphospholipid syndrome (APS) remains controversial. The purpose of this review is to provide updated recommendations. There is evidence that patients at risk of thrombosis are those with "a pattern of high risk antiphospholipid antibodies (aPL)" (presence of a lupus anticoagulant [LA], association of several aPL, or persistent aCL at a medium or high level), or those with associated systemic lupus erythematosus (SLE). The prescription of aspirin in primary prevention is recommended in SLE patients with positive LA or persistent aCL at a significant level. Secondary prevention is based on a very prolonged anticoagulation. An INR around 2.5 seems to be sufficient in patients with venous APS. In case of arterial events, the attitude is debated. We propose to maintain a target INR between 3 and 3.5. The possible occurrence of relapse despite anticoagulation in the therapeutic target may lead to the addition of aspirin. The development of new anti-thrombotic agents might change the management of APS in the coming years.


Apoptotic DNA fragmentation and tissue homeostasis.
DNA fragmentation is a hallmark of apoptosis. The tightly controlled activation of the apoptosis-specific endonucleases provides an effective means to ensure the removal of unwanted DNA and the timely completion of apoptosis. Over the past several years, crucial progress has been made in identifying the long-awaited apoptotic endonucleases, and their importance in tissue homeostasis is beginning to unfold. Here, we focus on the most recent discoveries about the functions and mechanisms of these endonucleases in the context of apoptosis. We also discuss consequences that defective DNA fragmentation might have for tissue homeostasis and disease development.


Pyoderma gangrenosum mimicking mediastinitis after cardiac surgery: a case study.
Pyoderma gangrenosum is a dermatosis which associates both, necrosis and polynuclear infiltration of the skin. While the aetiology is not well understood, the disease is thought to be due to immune system dysfunction and it can occur after minor trauma or surgery. Although it has seldom been reported after cardiac surgery in the literature, it is not exceptional. Here we report a case of pyoderma gangrenosum after coronary artery bypass grafting in a 76-year-old patient with chronic idiopathic myelofibrosis. Diagnosis was clinically made and the patient was treated with systemic steroids. The lesions showed a remarkable improvement with this therapy. In the field of cardiac surgery, physicians of the surgical team and nurses should think about this diagnosis in all rapidly expanding postoperative lesions without improvement after debridement or antibiotics. The authors have no conflicts of interest to declare.


[Role and action mechanisms of FZD5 in prostate cancer bone metastasis in mice].
To investigate the action mechanisms of the FZD5 gene in prostate cancer bone metastasis and search for some new treatments for this disease. We determined the expression level of the FZD5 gene in prostate cancer PC3 cells and, after transfection of siRNA into the PC3 cells and silence of the FZD5 gene, observed the changes in the migration and proliferation of the cells. We established the model of prostate cancer bone metastasis by tibial injection of prostate cancer cells in the nude mice. Then we injected control siRNA and FZD5-silenced siRNA into the tibia of the mice followed by evaluation of tumor-induced bone destruction by X-ray imaging at 0, 1, and 3 weeks and by HE staining at 3 weeks after injection. After transfection of FZD5-silenced siRNA into the prostate cancer PC3 cells, the expression of the FZD5 gene was decreased about 70%. The rate of cell proliferation was significantly lower in the gene silencing group than in the control (P < 0.05), and that of cell migration dropped by 30% in the former as compared with the latter group at 48 hours after FZD5 silencing (P < 0.05). At 3 weeks after injection of control siRNA or FZD5-silenced siRNA into the tibia of the mice, osteolytic damage was observed in both groups, though less in the FZD5 silencing group, with only a few remaining bone trabeculae visible. Silencing the FZD5 gene can reduce the migration and proliferation of prostate cancer cells, help to suppress bone metastasis and destruction, and thereby improve the survival rate and quality of life of the patients.


A comparison of hearing results following stapedotomy under local versus general anesthesia.
Advances in operative technique, instrumentation, and prosthesis design in otosclerosis surgery continue since Shea performed the first successful surgery. This is the first analysis to specifically compare post-operative hearing outcomes following stapedotomy surgery performed under local versus general anesthesia. Hearing outcomes were further stratified by comparing conventional perforator and Er:YAG laser ablation perforation techniques. Pre- and post-operative audiograms were retrospectively analyzed together with the method of anesthesia and the perforation technique for all patients with otosclerosis who underwent stapedotomy between 1998 and 2007. Pre-operative individual standard audiometry frequency thresholds (IFTs), air (AC) and bone conduction pure tone averages (PTA), and air bone gaps (ABG) were compared against post-operative results. Differences between pre- and post-operative PTAs and ABGs were compared between patients who received stapedotomy under local versus general anesthesia, as well as for patients who underwent conventional versus Er:YAG laser ablation perforations. Eighty-six patients were identified of which 24 % (n = 21) received local and 76 % (n = 65) received general anesthesia. Post-operative audiograms were available for 84 and 48 patients, respectively. Significant improvements were seen across all groups for standard 4-frequency AC-PTA and ABG and for IFTs up to 3 kHz. No significant difference was seen for IFTs between 4 and 6 kHz. A significant decline in post-operative hearing thresholds was seen at 8 kHz. Significant improvements in PTA and ABG were seen for all groups. There was a trend toward general compared to local anesthesia post-operative hearing results furthermore in combination with conventional perforation technique then with laser technique.


Cholecystoenteric anastomosis with the EEA stapler for cancer of the pancreas.
The integrity of a stapled anastomosis of the small bowel to the gallbladder with malignant distal biliary obstruction was investigated. Seven dogs were studied during a 52 week period following completion of a cholecystoenterostomy (Roux-Y) with an EEA stapler for bypass of a ligated common bile duct. A widely patent and secure anastomotic line was documented. Three human patients with unresectable cancer of the head of the pancreas who underwent stapled cholecystojejunostomies received excellent palliation as well. Cholecystojejunostomy with the EEA stapler is a rapid and safe procedure for unresectable, malignant distal biliary obstruction.


4-tert-butylcalix[4]arene having nitrile pendant groups as Hg²⁺ selective receptors.
We synthesized a series of 4-tert-butylcalix[4]arene nitriles (5-7) by the reaction of 4-tert-butylcalix[4]arene (1) with 4'-(6-bromohexyloxy)biphenyl-4-carbonitrile (2), 4'-(10-bromode-cyloxy)biphenyl-4-carbonitrile (3), and 4'-(12-bromododecyloxy)biphenyl-4-carbonitrile (4) and characterized their structures, respectively. The extraction abilities of newly synthesized 5-7 toward some selected heavy metal cations, such as Co(2+), Cu(2+), Ni(2+), Zn(2+), Cd(2+), and Hg(2+), were first evaluated and compared by the solvent extraction method. The extraction results revealed that 5-7 were efficient and selective cation receptors for Hg(2+) over-selected cations. On the other hand, the complexation behavior of Hg(2+) with 5-7 was also investigated by using NMR, UV-vis, and IR spectroscopic methods.


SILS cholecystectomy, early experience of a single institution: pilot study of 21 cases.
Many surgeons have attempted to reduce the number and the size of ports in laparoscopic cholecystectomy to decrease parietal trauma and pain, and to improve cosmetic results. We report a series of laparoscopic cholecystectomies using a single-port technique (SILS) through an umbilical incision in a pilot group of 21 cases. Our goal was to validate and develop the single-port access as a viable option. All the operations were performed using an umbilical SILS port (Ethicon). Most reported techniques utilize special purpose-made instruments. This article provides a stepwise description of the procedure using all straight instruments. No special reticulating instruments or flexible telescopes were used. We report our early difficulties and concerns about the procedure and propose solutions to the problems. Patients' request for improved cosmesis impels surgeons toward the application of SILS, but the true advantage of the technique should be assessed by more evidences. For this reason, we are planning a single-institution, prospective randomized controlled trial to compare postoperative pain, operating time and cosmetic result between one port and standard laparoscopic surgery.


Postoperative complications of capsulorhexis.
Capsulorhexis has become the preferred method of anterior capsulotomy, and untoward effects have not been frequently noted. Nevertheless, distinct complications of continuous tear anterior capsulotomy are now recognized. These include capsular bag hyperdistention, shrinkage of the anterior capsule opening with visual loss and/or intraocular lens decentration, and lens epithelial cell hyperproliferation on the posterior lens capsule. The latter has not been reported and may be associated wit reduction or closure of laser posterior capsulotomies in the form of a "string of pearls" around the capsule opening. Current styles of capsular surgery and intraocular lens implantation in which the anterior capsule edge overlaps the lens may be responsible for these phenomena. Although altering the surgical methods may obscure these complications, eliminating the anterior subcapsular and equatorial lens epithelial cells is necessary to prevent capsule contraction and lens epithelial cell hyperproliferation.


Transposon based activation tagging in diploid strawberry and monoploid derivatives of potato.
Diploid strawberry and potato transformed with a transposon tagging construct exhibited either global (strawberry) or local transposition (potato). An activation tagged, compact-sized strawberry mutant overexpressed the gene adjacent to Ds. As major fruit and vegetable crops, respectively, strawberry and potato are among the first horticultural crops with draft genome sequences. To study gene function, we examined transposon-tagged mutant strategies in model populations for both species, Fragaria vesca and Solanum tuberosum Group Phureja, using the same Activation/Dissociation (Ac/Ds) construct. Early somatic transposition during tissue culture occurred at a frequency of 18.5% in strawberry but not in potato transformants. Green fluorescent protein under a monocot promoter was a more reliable selectable marker in strawberry compared to potato. BASTA (gluphosinate herbicide) resistance served as an effective selectable marker for both species (80 and 85% reliable in strawberry and potato, respectively), although the effective concentration differed (0.5% for strawberry and 0.03% for potato). Transposons preferentially reinserted within genes (exons and introns) in both species. Real-time quantitative PCR revealed enhanced gene expression (670 and 298-fold expression compared to wild type in petiole and leaf tissue, respectively) for an activation tagged strawberry mutant with Ds inserted about 0.6 kb upstream from a gene coding for an epidermis-specific secreted glycoprotein EP1. Our data also suggested that endopolyploid (diploid) cells occurring in leaf explants of monoploid potato were the favored targets of T-DNA integration during transformation. Mutants obtained in these studies provide a useful resource for future genetic studies.


Glucosinolates and Isothiocyanates from Moringa oleifera: Chemical and Biological Approaches.
Alternative therapies, such as phytotherapy, are considered to improve the health status of people with chronic non-communicable diseases (CNCDs). In this regard, Moringa oleifera is currently being studied for its nutritional value and its total phenolic content. Besides phenolic compounds, the phytochemical composition is also of great interest. This composition is characterized by the presence of glucosinolates and isothiocyanates. Isothiocyanates formed by the biotransformation of Moringa glucosinolates contain an additional sugar in their chemical structure, which provides stability to these bioactive compounds over other isothiocyanates found in other crops. Both glucosinolates and isothiocyanates have been described as beneficial for the prevention and improvement of some chronic diseases. The content of glucosinolates in Moringa tissues can be enhanced by certain harvesting methods which in turn alters their final yield after extraction. This review aims to highlight certain features of glucosinolates and isothiocyanates from M. oleifera, such as their chemical structure, functionality, and main extraction and harvesting methods. Some of their health-promoting effects will also be addressed.


COVID-19, Climate Change, and Conflict in Honduras: A food system disruption analysis.
In Honduras, as in many settings between 2020 and 2022, food security was affected by the COVID-19 pandemic, climate change, and conflicts-what some refer to as "The Three Cs." These challenges have had overlapping impacts on food supply chains, food assistance programs, food prices, household purchasing power, physical access to food, and food acceptability. This article applies a food system disruption analysis-adapted from a fault tree analysis originally developed for a municipal context in the United States-to the context of Honduras to systematically examine how the Three Cs affected food availability, accessibility, and acceptability. This article demonstrates the value of approaching food security through a disruption analysis, especially for settings impacted by multiple, interconnected, ongoing crises.


Leadership in Education, Medical Education and Health.
We observe the impact of quality of leadership in our daily lives [...].


In vitro and In vivo Antioxidant Evaluation and Estimation of Total Phenolic, Flavonoidal Content of Mimosa pudica L.
Mimosa pudica Linn. (Mimosaceae) is traditionally used as a folk medicine to treat various ailments including convulsions, alopecia, diarrhea, dysentery, insomnia, tumor, wound healing, snake bite, etc., Here, the study was aimed to evaluate the antioxidant potential of M. pudica leaves extract against 2, 2-diphenyl-1-picrylhydrazyl (DPPH) (in vitro) and its modulatory effect on rat brain enzymes. Total phenolic, flavonoid contents, and in vitro antioxidant potential against DPPH radical were evaluated from various extracts of M. pudica leaves. In addition, ethyl acetate extract of Mimosa pudica leaves (EAMP) in doses of 100, 200, and 400 mg/kg/day were administered orally for 7 consecutive days to albino rats and evaluated for the oxidative stress markers as thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) from rat brain homogenate. The ethyl acetate extract showed the highest total phenolic content and total flavonoid content among other extracts of M. pudica leaves. The percentage inhibition and IC50 value of all the extracts were followed dose-dependency and found significant (P < 0.01) as compared to standard (ascorbic acid). The oxidative stress markers as SOD, CAT, and GSH were increased significantly (P < 0.01) at 200 and 400 mg/kg of EAMP treated animals and decreased significantly the TBARS level at 400 mg/kg of EAMP as compared to control group. These results revealed that the ethyl acetate extract of M. pudica exhibits both in vitro antioxidant activity against DPPH and in vivo antioxidant activity by modulating brain enzymes in the rat. This could be further correlated with its potential to neuroprotective activity due to the presence of flavonoids and phenolic contents in the extract. Total phenolic, flavonoid contents and in-vitro antioxidant potential were evaluated from various extracts of M. pudica leaves. Again, in-vivo antioxidant evaluation from brain homogenate on oxidative stress markers as TBARS, SOD, CAT and GSH from rat was investigated. Our findings revealed that M. pudica possesses both in-vitro and in-vivo antioxidant activity due to presence of phenolics and flavonoids.


Comparison of DNase, DNA-polymerase and RNA-polymerase activities present in the DNA-binding proteins of normal human dermis, epidermis, horny layer and psoriatic scales.
DNA-binding proteins (DBP) of normal human dermis, epidermis, horny layer and psoriatic scales represent a tissue-specific group of mostly nuclear nonhistone proteins. To analyse their function, the different DBP fractions were examined concerning the presence of DNase, DNA-polymerase and RNA-polymerase activities. DBP of normal epidermis and horny layer contain four different DNases. One DNase of both DBP fractions is active only at pH 5.0. Three DNases of epidermal DBP are active at a pH-range from 5.0--8.5, while the corresponding DNases of horny layer-DBP are most active at pH 7.4. Probably these DNases have changed their pH-optimum during keratinisation. DBP of psoriatic scales include no activity of these three DNases and the pH 5.0-DNases seem to have reduced DNA-affinity. Human dermis DBP contain quite another set of four DNases which hardly can be correlated to the DNases of epidermal DBP. DNA-polymerase activities are present in each fraction and derive from different DNA-polymerases. Two DNA-polymerases with pI-values of 4.5 and 9.3 may correspond to beta- and alpha-DNA-polymerase of eukaryotes, respectively. Further activity of proteins which are focussed at pH 6.5--7.2 and 8.2 could be detected. The proteins represent either tissue-specific DNA-polymerases or further thymidine monophosphate incorporating enzymes. Contrary, RNA-polymerase activity could not be enriched from correlating extracts by DNA-cellulose chromatography.


Apnea and seizures following retrobulbar local anesthetic injection.
Surgery on the eye is performed using topical anesthesia, retrobulbar anesthesia, peribulbar anesthesia, and general anesthesia. Retrobulbar anesthesia is associated with a number of complications that include apnea (respiratory arrest), seizures, or both. Although these complications are transient and self-limiting, they can be life-threatening if not recognized and treated early. We report two patients who developed apnea, one of whom had cardiorespiratory arrest; and two other patients who presented with seizures. We provided ventilation with 100% oxygen, treated the hypertension with nicardipine, and the tachycardia with esmolol. The patients did not have any residual complications.


Molecular variability of the Ancylostoma secreted Protein-2 (Aca-asp-2) gene from Ancylostoma caninum contributes to expand information on population genetic studies of hookworms.
Hookworm infection is a major public health problem in many regions of the world. Given the high levels of host morbidity and even mortality of the host caused by these infections, it is crucial to understand the genetic structure of hookworm populations. This understanding can provide insights into the ecology, transmission patterns, mechanisms of drug resistance, and the development of vaccines and immunotherapeutic strategies. Previously, we examined presumably neutral molecular markers, such as microsatellites and COI (Cytochrome C oxidase subunit 1) in Brazilian populations of Ancylostoma caninum. Here we analyze the molecular variability of a genomic fragment of the Aca-asp-2 (Ancylostoma secreted protein-2) gene from Ancylostoma caninum. This gene is a highly expressed and activated following the infection of the L3 larvae in the host. We obtained individuals of A. caninum from five different geographic locations in Brazil, sequenced and analyzed parts of the gene. The results revealed extensive polymorphism at this fragment, especially in the intronic region, indicating low selective pressure acting on these sequences. However, we also observed irregular distributions of nucleotides and polymorphisms in the coding region of this gene, resulting in the identification of 27 alleles. The data presented here contribute to expanding the understanding of population genetic studies of hookworms.


Laparoscopic excision of renal hydatid cyst in a preadolescent.
The traditional method of treating hydatid disease by open surgery has, in recent years, been challenged and minimally invasive approaches, including percutaneous and laparoscopic management, have been gaining wide acceptance. We present a 13-year-old boy with the diagnosis of isolated right renal hydatid disease, and a total pericystectomy was performed laparoscopically. Total operative time was 4.5 h and estimated blood loss was 150 ml. The only complication was an early urinary leak which was successfully treated with internal stenting. We believe further experience will reduce the operative time and complications.


How accurate is symptomatic and clinical evaluation of prolapse prior to surgical repair?
The aim of this study is to assess the accuracy of pre-operative evaluation of pelvic organ prolapse. The design is a prospective observational audit set at the gynaecology department, Teaching Hospital, UK. The population is composed of patients undergoing surgery for prolapse. One hundred and four patients admitted for prolapse surgeries were enrolled in the audit. Patients' notes were initially reviewed for adequacy of prolapse assessment in the clinic. Patients were then interviewed by the researchers and assessed using a validated Prolapse Quality of Life (P-QOL) questionnaire. The presence of unrecorded symptoms was noted. Prolapse examination in theatre under anaesthesia was compared to the findings in the clinic and the operation performed compared to the proposed operation. The outcome measures were as follows: (1) number of patients who had accurate prolapse symptom assessment before surgery when comparing clinical records with entries on P-QOL questionnaires; (2) number of patients having symptoms related to their pelvic organ prolapse that were not accurately assessed pre-operatively; and (3) the differences, if any, between pre-operative and intra-operative examination of prolapse. Sixteen patients in our cohort (15%) had adequate assessment of their prolapse pre-operatively. Symptoms that were not adequately assessed in descending order were the impact of prolapse on quality of life (76%), sexual function (75%), bowel function (27%) and lower urinary tract symptoms (12.5%). Thirty one patients (30%) had sexual dysfunction, 24 (23%) had bowel symptoms and 23 patients (22%) had urinary symptoms that were not recorded before surgery. Prolapse physical examination was adequate in 59% of the cases. Examinations in theatre were different from clinic findings in 38 cases (37%); 16 cases (42%) had a greater or lesser degree of prolapse than that described in the notes; and 11 cases (29%) had prolapse in a different compartment in the vagina. A combination of both (i.e. different degree of prolapse and prolapse in a different vaginal compartment) was found in another 11 cases (29%). The operation performed was different from the one proposed in the clinic in 21% of the cases (n=22). Clinical evaluation and examination of patients with vaginal prolapse is often inadequate. Prolapse physical examination in a clinic setting could be different from findings under anaesthesia. This can affect the operation to repair the prolapse. Patients should be counselled about this when listed for surgery.


Comprehensive Guide to Extracting and Expressing Fungal Secondary Metabolites with Aspergillus fumigatus as a Case Study.
Fungal secondary metabolites (SMs) have captured the interest of natural products researchers in academia and industry for decades. In recent years, the high rediscovery rate of previously characterized metabolites is making it increasingly difficult to uncover novel compounds. Additionally, the vast majority of fungal SMs reside in genetically intractable fungi or are silent under normal laboratory conditions in genetically tractable fungi. The fungal natural products community has broadly overcome these barriers by altering the physical growth conditions of the fungus and heterologous/homologous expression of biosynthetic gene cluster regulators or proteins. The protocols described here summarize vital methodologies needed when researching SM production in fungi. We also summarize the growth conditions, genetic backgrounds, and extraction protocols for every published SM in Aspergillus fumigatus, enabling readers to easily replicate the production of previously characterized SMs. Readers will also be equipped with the tools for developing their own strategy for expressing and extracting SMs from their given fungus or a suitable heterologous model system. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Making glycerol stocks from spore suspensions Alternate Protocol 1: Creating glycerol stocks from non-sporulating filamentous fungi Basic Protocol 2: Activating spore-suspension glycerol stocks Basic Protocol 3: Extracting secondary metabolites from Aspergillus spp grown on solid medium Alternate Protocol 2: Extracting secondary metabolites from Aspergillus spp using ethyl acetate Alternate Protocol 3: High-volume metabolite extraction using ethyl acetate Alternate Protocol 4: Extracting secondary metabolites from Aspergillus spp in liquid medium Support Protocol: Creating an overlay culture Basic Protocol 4: Extracting DNA from filamentous fungi Basic Protocol 5: Creating a DNA construct with double-joint PCR Alternate Protocol 5: Creating a DNA construct with yeast recombineering Basic Protocol 6: Transformation of Aspergillus spp Basic Protocol 7: Co-culturing fungi and bacteria for extraction of secondary metabolites.


miRNAs associated with prostate cancer risk and progression.
Prostate cancer is the most common malignancy among men in the US. Though considerable improvement in the diagnosis of prostate cancer has been achieved in the past decade, predicting disease outcome remains a major clinical challenge. Recent expression profiling studies in prostate cancer suggest microRNAs (miRNAs) may serve as potential biomarkers for prostate cancer risk and disease progression. miRNAs comprise a large family of about 22-nucleotide-long non-protein coding RNAs, regulate gene expression post-transcriptionally and participate in the regulation of numerous cellular processes. In this review, we discuss the current status of miRNA in studies evaluating the disease progression of prostate cancer. The discussion highlights key findings from previous studies, which reported the role of miRNAs in risk and progression of prostate cancer, providing an understanding of the influence of miRNA on prostate cancer. Our review indicates that somewhat consistent results exist between these studies and reports on several prostate cancer related miRNAs. Present promising candidates are miR-1, -21, 106b, 141, -145, -205, -221, and -375, which are the most frequently studied and seem to be the most promising for diagnosis and prognosis for prostate cancer. Nevertheless, the findings from previous studies suggest miRNAs may play an important role in the risk and progression of prostate cancer as promising biomarkers.


The effect of electrical field strength on activation and development of cloned caprine embryos.
The activation procedure used in nuclear transfer (NT) is one of the critical factors affecting the efficiency of animal cloning. The purpose of this study was to compare the effect of two electrical field strengths (EFS) for activation on the developmental competence of caprine NT embryos reconstructed from ear skin fibroblasts of adult Alpine does. The NT embryos were obtained by transfer of the quiescent fibroblasts at the fourth passage into the enucleated metaphase II (M II) oocytes. Four to five hours after electrical fusion, the NT-embryos were activated by EFS either at 1.67 or at 2.33 kV/cm and immediately incubated in 6-DMAP (2 mM) for 4 h. The cleavage rate of the NT-embryos activated with 2.33 kV/cm was greater than that activated with 1.67 kV/cm after in vitro culture for 18 h (65.6% versus 19.6%, p < 0.001). No pregnancy was found in 14 recipient does after transferring 51 NT embryos at 1-2 cell stages activated with 1.67 kV/cm. In contrast, two of the seven recipients were pregnant and gave birth to three kids after transferring 61 NT embryos at 1-2 cell stages activated by 2.33 kV/cm. The birth weights of three cloned kids were within the normal range of Alpine goats. However, one kid died 1h after birth while the remaining two are still healthy. DNA analysis by polymerase chain reaction (single-strand conformation polymorphism, SSCP) confirmed that the three kids were genetically identical to the nuclear donor.


Structural characterization and immunogenicity in wild-type and immune tolerant mice of degraded recombinant human interferon alpha2b.
This study was conducted to study the influence of protein structure on the immunogenicity in wild-type and immune tolerant mice of well-characterized degradation products of recombinant human interferon alpha2b (rhIFNalpha2b). RhIFNalpha2b was degraded by metal-catalyzed oxidation (M), cross-linking with glutaraldehyde (G), oxidation with hydrogen peroxide (H), and incubation in a boiling water bath (B). The products were characterized with UV absorption, circular dichroism and fluorescence spectroscopy, gel permeation chromatography, reverse-phase high-pressure liquid chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blotting, and mass spectrometry. The immunogenicity of the products was evaluated in wild-type mice and in transgenic mice immune tolerant for hIFNalpha2. Serum antibodies were detected by enzyme-linked immunosorbent assay or surface plasmon resonance. M-rhIFNalpha2b contained covalently aggregated rhIFNalpha2b with three methionines partly oxidized to methionine sulfoxides. G-rhIFNalpha2b contained covalent aggregates and did not show changes in secondary structure. H-rhIFNalpha2b was only chemically changed with four partly oxidized methionines. B-rhIFNalpha2b was largely unfolded and heavily aggregated. Nontreated (N) rhIFNalpha2b was immunogenic in the wild-type mice but not in the transgenic mice, showing that the latter were immune tolerant for rhIFNalpha2b. The anti-rhIFNalpha2b antibody levels in the wild-type mice depended on the degradation product: M-rhIFNalpha2b > H-rhIFNalpha2b approximately N-rhIFNalpha2b >> B-rhIFNalpha2b; G-rhIFNalpha2b did not induce anti-rhIFNalpha2b antibodies. In the transgenic mice, only M-rhIFNalpha2b could break the immune tolerance. RhIFNalpha2b immunogenicity is related to its structural integrity. Moreover, the immunogenicity of aggregated rhIFNalpha2b depends on the structure and orientation of the constituent protein molecules and/or on the aggregate size.


A risk-based sensor placement methodology.
A risk-based sensor placement methodology is proposed to solve the problem of optimal location of sensors to protect population against the exposure to, and effects of, known and/or postulated chemical, biological, and/or radiological threats. Risk is calculated as a quantitative value representing population at risk from exposure at standard exposure levels. Historical meteorological data are used to characterize weather conditions as the frequency of wind speed and direction pairs. The meteorological data drive atmospheric transport and dispersion modeling of the threats, the results of which are used to calculate risk values. Sensor locations are determined via an iterative dynamic programming algorithm whereby threats detected by sensors placed in prior iterations are removed from consideration in subsequent iterations. In addition to the risk-based placement algorithm, the proposed methodology provides a quantification of the marginal utility of each additional sensor. This is the fraction of the total risk accounted for by placement of the sensor. Thus, the criteria for halting the iterative process can be the number of sensors available, a threshold marginal utility value, and/or a minimum cumulative utility achieved with all sensors.


Apoptosis in CHO cell batch cultures: examination by flow cytometry.
Chinese hamster ovary cells grown under conditions which are optimal for the production of a genetically engineered protein in batch culture, lose significant viability shortly after entering the stationary phase. This cell death was investigated morphologically and was found to be almost exclusively via apoptosi. Furthermore, cells were analyzed by flow cytometry using a fluorescent DNA end-labeling assay to label apoptotic cells, in conjunction with cell cycle analysis using propidium iodide. Apoptotic cells could be detected by this method, and by the radioactive end-labeling of extracted DNA, on all days of culture from day 1 to day 7; however, the degree of apoptotic cell death increased dramatically when the cells entered the stationary phase, rising to 50-60% of the total cell number at the termination of the culture. Flow cytometric analysis showed that the majority of cells underwent apoptosis whilst in G(1)/G(0) and formed an apoptotic population with high DNA FITC end-labeling and hypodiploid propidium iodide binding. Additionally, the ability or inability to secrete specific protein products did not appear to interfere with the development of the apoptotic population with time.


The non-conserved region of cucumopine-type Agrobacterium rhizogenes T-DNA is responsible for hairy root induction.
The T-DNA regions of three strains of Ri plasmids 1855, 8196, 2659 (agropine, mannopine and cucumopine type respectively) share two highly conserved regions flanking a non-homologous central part. We have cloned segments of the cucumopine Ri plasmid 2659 T-DNA in the binary vector system Bin 19 and infected carrot discs with recombinant Agrobacterium strains. We show here that the central non-conserved region is crucial in hairy root induction as it is sufficient to induce rooting on the apical (auxin-rich) surface of carrot discs; in order to observe rooting on the basal (auxin-depleted) side of the discs, a longer T-DNA fragment, also encompassing part of the right conserved region, had to be utilized in conjunction with a Agrobacterium strain carrying aux genes. Differences of growth properties in culture are exhibited by roots transformed with different fragments of pRi 2659 T-DNA, although all transformed roots show the plagiotropic behaviour typical of hairy roots.


Deep learning for automatic diagnosis of gastric dysplasia using whole-slide histopathology images in endoscopic specimens.
Distinguishing gastric epithelial regeneration change from dysplasia and histopathological diagnosis of dysplasia is subject to interobserver disagreement in endoscopic specimens. In this study, we developed a method to distinguish gastric epithelial regeneration change from dysplasia and further subclassify dysplasia. Meanwhile, optimized the cross-hospital diagnosis using domain adaption (DA). 897 whole slide images (WSIs) of endoscopic specimens from two hospitals were divided into training, internal validation, and external validation cohorts. We developed a deep learning (DL) with DA (DLDA) model to classify gastric dysplasia and epithelial regeneration change into three categories: negative for dysplasia (NFD), low-grade dysplasia (LGD), and high-grade dysplasia (HGD)/intramucosal invasion neoplasia (IMN). The diagnosis based on the DLDA model was compared to 12 pathologists using 100 gastric biopsy cases. In the internal validation cohort, the diagnostic performance measured by the macro-averaged area under the receiver operating characteristic curve (AUC) was 0.97. In the independent external validation cohort, our DLDA models increased macro-averaged AUC from 0.67 to 0.82. In terms of the NFD and HGD cases, our model's diagnostic sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were significantly higher than junior and senior pathologists. Our model's diagnostic sensitivity, NPV, was higher than specialist pathologists. We demonstrated that our DLDA model could distinguish gastric epithelial regeneration change from dysplasia and further subclassify dysplasia in endoscopic specimens. Meanwhile, achieved significant improvement of diagnosis cross-hospital.


Measurement of nonlinear property of artery wall using remote cyclic actuation.
For tissue characterization of the arterial wall, we developed a "phased tracking" method to measure the strain (change in wall thickness) and elasticity of the arterial wall. To improve the accuracy of tissue characterization, we are now attempting to measure other mechanical properties in addition to elasticity. In this study, the change in elasticity during the cardiac diastole was measured with ultrasound by generating a change in internal pressure using remote cyclic actuation. From the measured change in elasticity during cardiac diastole, the nonlinear property in the stress-strain relationship of the artery wall was estimated. In basic experiments using a silicone rubber tube and in vivo experiments in human carotid arteries. The proposal method enables the noninvasive measurement of the nonlinear mechanical property in addition to the elasticity of the arterial wall.


Co-composting of municipal solid waste mixed with matured sewage sludge: The relationship between N2O emissions and denitrifying gene abundance.
Aerobic composting is an alternative measure to the disposal of municipal solid waste (MSW). However, it produces nitrous oxide (N2O), a highly potent greenhouse via microbial nitrification and denitrification. In this study, the effects of matured sewage sludge (MSS) amendment on N2O emissions and the inter-relationships between N2O emissions and the abundance of denitrifying bacteria were investigated during aerobic composting of MSW. The results demonstrated that MSW composting with MSS amendments (C1, and C2, with a MSW to MSS ratio of 2:1 and 4:1, (v/v), respectively) significantly increased N2O emissions during the initial stage, yet contributed to the mitigation of N2O emissions during the cooling and maturation stage. MSS amended composting emitted a total of 18.4%-25.7% less N2O than the control treatment without MSS amendment (CK). Matured sewage sludge amendment also significantly altered the abundance of denitrifying bacteria. The quantification of denitrifying functional genes revealed that the N2O emission rate had a significant positive correlation with the abundance of the nirS, nirK genes in both treatments with MSS amendment. The nosZ/(nirS + nirK) ratio could be a good indicator for predicting N2O emissions. The higher N2O emission rate during the initial stage of composting mixed with MSS was characterized by lower nosZ/(nirS + nirK) ratios, compared to CK treatment. Higher ratios of nosZ/(nirS + nirK) were measured during the cooling and maturation stage in treatments with MSS which resulted in a reduction of the N2O emissions. These results demonstrated that MSS amendment could be a valid strategy for mitigating N2O emissions during MSW composting.


Microfibrous substrate geometry as a critical trigger for organization, self-renewal, and differentiation of human embryonic stem cells within synthetic 3-dimensional microenvironments.
Substrates used to culture human embryonic stem cells (hESCs) are typically 2-dimensional (2-D) in nature, with limited ability to recapitulate in vivo-like 3-dimensional (3-D) microenvironments. We examined critical determinants of hESC self-renewal in poly-d-lysine-pretreated synthetic polymer-based substrates with variable microgeometries, including planar 2-D films, macroporous 3-D sponges, and microfibrous 3-D fiber mats. Completely synthetic 2-D substrates and 3-D macroporous scaffolds failed to retain hESCs or support self-renewal or differentiation. However, synthetic microfibrous geometries made from electrospun polymer fibers were found to promote cell adhesion, viability, proliferation, self-renewal, and directed differentiation of hESCs in the absence of any exogenous matrix proteins. Mechanistic studies of hESC adhesion within microfibrous scaffolds indicated that enhanced cell confinement in such geometries increased cell-cell contacts and altered colony organization. Moreover, the microfibrous scaffolds also induced hESCs to deposit and organize extracellular matrix proteins like laminin such that the distribution of laminin was more closely associated with the cells than the Matrigel treatment, where the laminin remained associated with the coated fibers. The production of and binding to laminin was critical for formation of viable hESC colonies on synthetic fibrous scaffolds. Thus, synthetic substrates with specific 3-D microgeometries can support hESC colony formation, self-renewal, and directed differentiation to multiple lineages while obviating the stringent needs for complex, exogenous matrices. Similar scaffolds could serve as tools for developmental biology studies in 3-D and for stem cell differentiation in situ and transplantation using defined humanized conditions.


Quantitative evaluation of cerebral hemodynamics in patients with moyamoya disease by dynamic susceptibility contrast magnetic resonance imaging--comparison with positron emission tomography.
We examined whether the degree of hemodynamic stress in patients with chronic occlusive cerebral vascular disease can be quantitatively evaluated with the use of perfusion-weighted magnetic resonance imaging (PWI). Thirty-six patients with moyamoya disease (mean age, 26.8 years; range, 18 to 59) underwent PWI and positron emission tomography (PET) within a month's interval. The PWI data were calculated by three different analytic methods. The cerebral blood flow (CBF) ratio, cerebral blood volume (CBV) ratio, and mean transit time (MTT) of the anterior circulation were calculated using the cerebellum as a control region and compared with PET data on the same three parameters and oxygen extraction fraction (OEF). Parametric maps of PWI attained a higher resolution than the PET maps and revealed focal perfusion failure on a gyrus-by-gyrus level. The relative CBV and MTT obtained with PWI showed significant linear correlations with the corresponding PET values (CBV, R2 = 0.47 to 0.58; MTT, R2 = 0.32 to 0.68). We also found that we could detect regions with abnormally elevated OEF and CBV based on the delay of PWI-measured MTT relative to the control region by defining a 2.0-sec delay as a threshold. The sensitivity and specificity were 92.3% and 100% in detecting regions with abnormally elevated OEF, and 20.0% and 100% in detecting regions with abnormally elevated CBV, respectively. Among the parameters obtained with PWI, our results suggested that the relative CBV value and delay of MTT might be quantitatively manipulated to assist in clinical decision-making for patients with moyamoya disease.


yst gene expression in Yersinia enterocolitica is positively regulated by a chromosomal region that is highly homologous to Escherichia coli host factor 1 gene (hfq).
Yersinia enterocolitica produces heat-stable enterotoxin (Y-ST) as one of its virulence factors. The yst gene, however, frequently and spontaneously becomes inactive (silent) during storage, which is accompanied by concurrent changes in some biological properties such as colony morphology, growth rate, carbon fermentation and ornithine decarboxylase activity. Northern blot analysis revealed that the level of mRNA for yst was repressed. To investigate the regulatory region, we transformed a yst-silent strain with a chromosomal gene library of Y-ST producing an isogenic counterpart. Out of 3604 clones, one clone resumed the Y-ST production and concurrently other biological properties. An open reading frame in this clone was designated as yrp, yersinia regulator for pleiotropic phenotype. Deduced from the nucleotide sequence, Yrp was a small protein of I01 amino acids with no similarity with any regulatory factor described, but showed high homology with an Escherichia coli host factor 1 required for Q beta-replicase, and with an Azorhizobium caulinodans NrfA required for the expression of nifA. The yrp gene mutation caused decreased negative supercoiling of plasmids, as did hfq. The yrp gene could similarly complement Y-ST production in two other silent strains of Y. enterocolitica. In all three silent strains examined, we found various mutations in the yrp region.


Osseointegrated craniofacial implants in the rehabilitation of orbital defects: an update of a retrospective experience in the United States.
Since their introduction, craniofacial implants have been used in prosthetic rehabilitation of facial defects. The literature, however, indicates marked variability in outcomes using implants for the retention of orbital prostheses. A multicenter report updating the experience in the United States with the use of craniofacial implants for prosthetic rehabilitation of orbital defects is presented. Surveys were sent to clinicians at 25 centers where maxillofacial prosthetic treatment is provided to obtain retrospective data regarding patients who completed implant-retained orbital prosthetic rehabilitation. Data on implant placement location, radiation treatment history, and use of hyperbaric oxygen therapy were collected and assessed in relationship to implant survival over time. The Kaplan-Meier life table and Wilcoxon analyses (alpha = .05) were used to assess the significance of the findings. Ten centers responded, providing data suitable for statistical analysis on 153 implants placed to retain 44 orbital prostheses and followed for a mean period of 52.6 months. Forty-one implant integration failures occurred during this follow-up period, resulting in an overall integration survival rate of 73.2%. No significant relationship was found between radiation treatment history, hyperbaric oxygen therapy history, or implant placement location and implant survival. Individual responses revealed large variability between reporting centers in treatment outcomes. Craniofacial implants may offer marked benefits in the prosthetic rehabilitation of orbital defects when compared to conventional adhesive retention designs. However, questions remain regarding long-term predictability and the impact specific factors may have on treatment outcomes. Insufficient data is currently available from which to draw statistically meaningful conclusions. The establishment of a national database designed to acquire adequate data to assess treatment outcomes is recommended.


Transcatheter Treatment of Functional Mitral Regurgitation in Patients with Heart Failure: The COAPT Trial.
Secondary (functional) mitral regurgitation is strongly associated with recurrent heart failure (HF) hospitalizations, poor quality of life, and high rates of mortality. The COAPT trial demonstrated that transcatheter edge-to-edge mitral leaflet repair with the MitraClip device led to a decrease in the severity of secondary mitral regurgitation, a significantly lower rate of hospitalization for heart failure, lower mortality, and better quality of life and functional capacity within 24 months of follow-up compared with medical therapy alone. In this article, the authors review the COAPT trial rationale, design, results, and their clinical implications.


Involvement of adenosine A2A receptors in the induction of c-fos expression by clozapine and haloperidol.
Acute administration of the atypical antipsychotic clozapine induced a regional pattern of c-fos expression characterized by an increase in Fos-like-immunoreactivity (FLI) in the prefrontal and prelimbic/infralimbic cortices, nucleus accumbens, and lateral septum and a weak activation of FLI in the striatum. Haloperidol, similarly to clozapine, increased FLI in the nucleus accumbens and lateral septum, but it did not induce FLI in prefrontal and prelimbic/infralimbic cortices. Moreover, haloperidol increased FLI in the striatum. To gain insight into the mechanism by which clozapine and haloperidol induced FLI in these brain structures, we evaluated whether blockade of adenosine A2A receptors could influence these effects. The selective and high-affinity A2A receptor antagonist SCH 58261 (5 mg/kg) completely abolished FLI induced by clozapine (20 mg/kg) in all subdivisions of the nucleus accumbens (rostral pole, shell and core) and striatum, but did not affect the number of Fos-like positive neurons in the prefrontal, prelimbic/infralimbic cortices, and lateral septum. SCH 58261 (5 mg/kg) reduced FLI induced by haloperidol (0.1 mg/kg) in the striatum, lateral septum, and all nucleus accumbens subdivisions. In contrast, FLI induced by 0.5 mg/kg of haloperidol in the shell and core of the nucleus accumbens was not affected by SCH 58261. The results show that adenosine A2A receptors participate in the induction of FLI by clozapine and haloperidol and support the concept that A2A receptors are involved in the mediation of antipsychotic effects.


Inhibition of [3H]thymidine incorporation by Mycoplasma arginini-infected cells due to enzymatic cleavage of the nucleoside.
Culture supernatants contaminated by Mycoplasma arginini inhibit the incorporation of [3H]thymidine ([3H]dThd) by cytotoxic T lymphocyte cell lines. This study presents evidence that the inhibition of uptake of the nucleoside is due to the rapid cleavage of the exogenous [3H]dThd into thymine. Uridine and cytidine as well as dThd are degraded by the mycoplasma-contaminated supernatants, while no cleavage was observed with uninfected supernatants. Cells contaminated by mycoplasma apparently release a pyrimidine-specific nucleosidase, possibly a dThd phosphorylase, which is responsible for the inhibition.


Experimental autoimmune uveoretinitis in athymic rats: specific IgE response to retinal S-antigen and disease.
Experimental autoimmune uveoretinitis (EAU) is an ocular autoimmune disease induced in rats by immunization with retinal S-antigen. Athymic nude rats (rnu/rnu) have been previously shown to be refractory to EAU induction and antibody production to S-antigen, while heterozygous (rnu/+) are good responders. Increasing the antigen dose and adding pertussis adjuvant produced ocular disease in some nude rats, and antibody response in most of them. Specific IgE antibodies were demonstrated by ELI-SA only in the serum of nude rats presenting the disease. However, most immunized nude rats had evidence of mast cell sensitization to S-antigen (direct degranulation test) and of circulating specific IgE detected by passive sensitization of normal mast cells (indirect degranulation test). This positive response could be explained by an incomplete depletion of the different T lymphocyte subsets.


[Treatments and diagnosis of lung cancer--field study results: report from japan lung cancer TNM classification committee].
The results of 4,931 lung cancer cases treated during 1970-72 were examined to evaluate the factors (especially N factors) influencing the prognosis of lung cancer. The background factors examined included T-factor, N-factor, M-factor, clinical stage, histological type, age and sex. The prognosis was also examined according to treatment method. The 5-year survival rate in the curative resection only group was 42.6%, in curative resection + chemotherapy 52.1%, and in curative resection + chemotherapy 52.1%, and 54.8%. The results suggest improved 5-year survival by multimodality treatment.


Acaricidal activity of the organic extracts of thirteen South African plants against Rhipicephalus (Boophilus) decoloratus (Acari: Ixodidae).
The African blue tick, Rhipicephalus (Boophilus) decoloratus, is a common tick species found in South Africa and affects cattle production as well as vectoring pathogens in regions of Africa and Asia. In an attempt to develop a non-toxic, lower cost and environmentally friendly tick control method, twenty-six plant extracts were prepared from thirteen plant species using 99.5% acetone and 99% ethanol. The adapted Shaw Larval Immersion Test (SLIT) was used to test the efficacy of the extracts. A 1% solution of each of the plant extracts was prepared for efficacy testing and the ethanol extracts were found to have better acaricidal activity than the acetone extracts. The ethanol extract from the leaves and flowers of Calpurnia aurea had the best activity [corrected mortality (CM)=82.9%] which was followed by the stem extract of Cissus quadrangularis (CM=80.4%). The plant species were screened against Vero cells and were found to have low toxicity. From this study it is apparent that there is potential for the development of botanicals as natural acaricides against R. (B.) decoloratus.


Viral diversity limits immune diversity in asymptomatic phase of HIV infection.
We propose a new diversity threshold theory which states that the specific CTLs to the viral strain become inactivated (that is, some HIV strain can escape from its specific immune response) when the diversity of HIV strains exceeds some threshold number. We call this number "immune diversity threshold". Our theory can explain the inactivation of specific immune response and a limit of maximum immune diversity. We can conclude that the accumulation of viral diversity eventually leads to AIDS.


Neuropathology of secondary generalized epilepsy--(Lennox-Gastaut syndrome)--a case report.
Neuropathological considerations were performed on a case, who went into Lennox syndrome after an acute encephalopathy at the infantile period, and moreover who fell into an akinetic-mute state derived from brain damage by herniation caused by a head injury and subsequent status epilepticus. Neuropahtological background in the present case of Lennox syndrome is thought to be based on the widespread unilateral cerebral lesions and the basal ganglional, especially thalamic, degenerations derived secondarily from the diffuse cerebral damage. The patient revealed akinetic mutism with the disappearance of the epileptic seizures and the desynchronization of the EEG's, when the brain lesions formed at the adult period spread over the opposite hemispheric limbic system and the brain stem tectum.


Efficient isolation of membrane-associated exopolysaccharides of four commercial bifidobacterial strains.
Bifidobacteria confer many health effects, such as fiber digestion, pathogen inhibition and immune system maturation, especially in the newborn infant. The bifidobacterial exopolysaccharides (EPS) are often associated with important health effects, but their thorough investigation is hampered by lack of knowledge of the EPS localization, which is important for efficient EPS isolation. Here we present a straightforward isolation procedure to obtain EPS of four commercial bifidobacterial strains (B. adolescentis, B. bifidum, B. breve, and B. infantis), that are localized at the cell membrane (evidenced using cryo-EM). This procedure can be applied to other bifidobacterial strains, to facilitate the easy isolation and purification for biological experiments and future application in nutraceuticals. In addition, we demonstrate structural differences in the EPS of the four bifidobacterial strains, in terms of monosaccharide composition and size, highlighting the potential of the isolated EPS for determining specific structure-activity effects of bifidobacteria.


Effect of cyclic AMP on catabolite repressed bacterial sporogenesis of an anaerobe.
The sporulation of a high frequency sporogenic mutant of Clostridium botulinum was reduced to less than 30% in a medium containing 270 mM glucose. The repression was reversed from 30 to greater than 80% sporulation by the addition of 10(-5) or 10(-4) M cyclic 3',5'-adenosine monophosphate (cAMP) or monobutyrul cyclic AMP (B-cAMP). No difference was observed in amount of growth with the addition of either the cAMP or B-cAMP. Glucose consumption was enhanced by the addition of either of the cyclic nucleotides and corresponding changes in pH were observed. The catabolite repression by glucose was reversed by ATP or ADP. Except for GTP, guanine nucleotides were not effective. The intracellular cyclic AMP levels were high in vegetative, sporulating and derepressed cells, but low in glucose-repressed cells. The findings suggest that the sporulation of the anaerobe was sensitive to catabolite repression which was specifically reversed by cyclic AMP.


Li5 SnP3 - a Member of the Series Li10+4x Sn2-x P6 for x=0 Comprising the Fast Lithium-Ion Conductors Li8 SnP4 (x=0.5) and Li14 SnP6 (x=1).
The targeted search for suitable solid-state ionic conductors requires a certain understanding of the conduction mechanism and the correlation of the structures and the resulting properties of the material. Thus, the investigation of various ionic conductors with respect to their structural composition is crucial for the design of next-generation materials as demanded. We report here on Li5 SnP3 which completes with x=0 the series Li10+4x Sn2-x P6 of the fast lithium-ion conductors α- and β-Li8 SnP4 (x=0.5) and Li14 SnP6 (x=1). Synthesis, crystal structure determination by single-crystal and powder X-ray diffraction methods, as well as 6 Li, 31 P and 119 Sn MAS NMR and temperature-dependent 7 Li NMR spectroscopy together with electrochemical impedance studies are reported. The correlation between the ionic conductivity and the occupation of octahedral and tetrahedral sites in a close-packed array of P atoms in the series of compounds is discussed. We conclude from this series that in order to receive fast ion conductors a partial occupation of the octahedral vacancies seems to be crucial.


Comprehensive Analysis Identifies COL1A1, COL3A1, and POSTN as Key Genes Associated with Brain Metastasis in Patients with Breast Cancer.
Brain metastasis (BM) is associated with a high mortality in patients with breast cancer (BC). Nevertheless, the molecular mechanisms of BM in BM remain uncertain. The study aims to identify the key genes in BC in relation to BM and to assess their prognostic value. Two microarray datasets GSE125989 and GSE100534 were downloaded from the Gene Expression Omnibus (GEO) database to identify differentially expressed genes (DEGs) between primary BC and BM samples. The function enrichment analysis and protein-protein interaction (PPI) network were performed. We mapped hub genes into the Kaplan-Meier database for their correlations with BC survival. Venn diagram analysis showed an overlapped upregulated DEG and 18 overlapped downregulated ones between primary BC and BM samples. We constructed the PPI network, and top 5 hub genes were sorted out according to the node degree, including type I collagen α 1 chain (COL1A1), lumican (LUM), type III collagen α 1 chain (COL3A1), type V collagen α 2 chain (COL5A2), and periosteal protein (POSTN). The Kaplan-Meier database analysis found that COL1A1, COL3A1, and POSTN were significantly correlated with overall survival of BC patients. The study suggests that COL1A1, COL3A1, and POSTN may be key genes associated with BM in patients with BC.


Effect of ultrasound image enhancement software on the quality of vision of regional anesthesia needles.
Our objective was to evaluate and compare the visualization of different types of needles with or without ultrasound image enhancement software, both in biological tissues and artificial models. This is an observational study on fresh porcine tissue and gelatin models. Six types of plexus needles were studied. The same anesthesiologist performed in-plane punctures with each needle at 30°, 40° and 50° in both 2D mode and using software-based enhanced mode without changing position, generating 72 images. The images were evaluated blind by 38 anesthesiologists with at least two years of experience in ultrasound and rated from 0 to 10. A univariate and multivariate analysis was performed to identify differences between the images according to needle, mode, angle and experimental model. We described the results as mean (standard deviation). The Echoplex needle, 8.31 (1.94), was significantly better than the Sonoplex needle, 7.53 (2.16), P=0.0003, and both were significantly better than the other needles (P<0.0001). Significant differences were also found in favor of the gelatin model, 7.26 (2.48) vs. 6.24 (3.67), P<0.0001, and with ultrasound image enhancement software, 8.59 (1.55) vs. 4.91 (3.31), P<0.0001. These differences were confirmed by multivariate analysis. Although there are differences between the different types of needles used with ultrasound visualization strategies, ultrasound image enhancement software provides good visualization, regardless of the model chosen.


Assessment of protein dynamics and DNA repair following generation of DNA double-strand breaks at defined genomic sites.
The formation of protein aggregates (foci) at sites of DNA double-strand breaks (DSBs) is mainly studied by immunostaining and is hence limited by the low resolution of light microscopy and the availability of appropriate and selective antibodies. Here, we describe a system using enzymatic creation of site-specific DNA DSBs within the human genome combined with chromatin immunoprecipitation (ChIP) that enables molecular probing of a DSB. Following induction of the I-PpoI enzyme and generation of DSBs, cellular DNA and proteins are crosslinked and analyzed by ChIP for specific proteins at the site of the break. The system allows the direct detection of protein and chromatin dynamics at the site of the break with high resolution, as well as direct measurement of DNA repair defects in human cells. Starting with fragmented chromatin, results can be achieved in 2-3 d.


Solving inverse problems for biological models using the collage method for differential equations.
In the first part of this paper we show how inverse problems for differential equations can be solved using the so-called collage method. Inverse problems can be solved by minimizing the collage distance in an appropriate metric space. We then provide several numerical examples in mathematical biology. We consider applications of this approach to the following areas: population dynamics, mRNA and protein concentration, bacteria and amoeba cells interaction, tumor growth.


Tumor-stroma interaction: positive feedback regulation of extracellular matrix metalloproteinase inducer (EMMPRIN) expression and matrix metalloproteinase-dependent generation of soluble EMMPRIN.
Matrix metalloproteinases (MMPs) are metal-dependent endopeptidases that play pivotal roles in tumor disease progression. In many solid tumors, MMPs are indeed produced by tumor stromal cells, rather than by tumor cells. This expression pattern is, at least in part, regulated by tumor-stroma interaction via tumor cell-associated extracellular matrix metalloproteinase inducer (EMMPRIN). In vitro, recombinant EMMPRIN dose-dependently stimulated MMP-1 production by primary human fibroblast cells. Interestingly, in addition to stimulating MMP expression, EMMPRIN also induced its own gene expression. To further explore this potential positive feedback regulatory mechanism, we generated human breast cancer cells expressing different levels of EMMPRIN. Coculture of EMMPRIN-positive tumor cells with fibroblast cells resulted in a concomitant stimulation of MMP-2, MMP-9, and EMMPRIN production. This induction was EMMPRIN dependent, was further enhanced by overexpression, and was reduced by antisense suppression of EMMPRIN expression in tumor cells. Increased expression of membrane-associated EMMPRIN was accompanied by an MMP-dependent generation of a soluble form of EMMPRIN representing a proteolytic cleavage product lacking the carboxyl terminus. On the basis of these findings, we propose a model in which tumor cell-associated EMMPRIN stimulates MMPs, as well as EMMPRIN expression in tumor stroma. Increased MMP activity in tumor local environment results in proteolytic cleavage of membrane-associated EMMPRIN, releasing soluble EMMPRIN. Soluble EMMPRIN in turn acts in a paracrine fashion on stroma cells that are both adjacent and distant to tumor sites to further stimulate the production of MMPs and additional EMMPRIN, which consequently contributes to tumor angiogenesis, tumor growth, and metastasis.


A fluorimetric assay for cortisol.
A simple, rapid and sensitive fluorimetric assay for the quantitative determination of cortisol is reported. The assay is based on the formation of a fluorescent dye when cortisol is incubated with a mixture of sulfuric acid and acetic acid. The fluorescence spectrum recorded for the resulting dye shows a maximum extinction at 475 nm and a maximum emission at 525 nm. The solvent 2-methyl-4-pentanone was used for extraction and was found to act as a fluorescence amplifier. A limit of detection of 2.7 muM was achieved, making it possible to forego solvent evaporation. The assay suffers minor interference from 11-deoxycortisol which exhibits low fluorescence at lambda (ex): 460 nm; lambda (em): 505 nm. Typical standard deviations were below 4%. We validated the assay using a biotransformation with recombinant Schizosaccharomyces pombe which regioselectively hydroxylates 11-deoxycortisol to cortisol. The method described herein is suitable for preliminary screening of microorganisms capable of steroid hydroxylation.


Virtual reality tour for first-time users of highly automated cars: Comparing the effects of virtual environments with different levels of interaction fidelity.
Research in aviation and driving has highlighted the importance of training as an effective approach to reduce the costs associated with the supervisory role of the human in automated systems. However, only a few studies have investigated the effect of training on highly automated driving. Moreover, available interactive trainings are mostly based on automated driving simulators and the application of immersive technology such as Virtual Reality (VR) as a low-cost training solution has not been widely adopted. In this study, we developed three types of familiarization tours (low-fidelity VR, high-fidelity VR, and video) to train first-time users of highly automated cars. Then, the effectiveness of these tours was investigated on automation trust and driving performance in several critical and non-critical transition tasks in four groups: control, video, low-fidelity VR, and high-fidelity VR. The results revealed the positive impact of the tours on trust and transition performance at the first time of measurement. Takeover quality only improved when practices were presented in high-fidelity VR. After three times of exposure to transition requests, trust and transition performance of all groups converged to those of the high-fidelity VR group, demonstrating that: a) experiencing takeover transition during the training may reduce costs associated with first critical takeover request in highly automated driving, b) the VR tour with high level of interaction fidelity was superior to other training methods, and c) untrained and less-trained drivers learned about automation after a few trials. Knowledge resulting from this research could help develop cost-effective solutions for automated driving training in dealerships and car rental centers.


3D anatomy and deformation of the seated buttocks.
To describe the 3D anatomy and deformation of the buttocks during sitting. The buttocks of 4 able-bodied individuals and 3 individuals with spinal cord injury were scanned sitting in a FONAR Upright MRI. T1-weighted Fast Spin Echo scans were collected with the individuals seated on a custom wheelchair cushion to unload the ischial tuberosities (ITs) and seated on a 3 inch foam cushion. Multi-planar scans were analyzed, and the muscle, bone and adipose tissue was manually segmented for 3D rendering and analysis of the quantity, geometry, and location of tissues. The gluteus maximus was positioned lateral and posterior to the IT, covering the inferior portion of the IT for only 2 able-bodied participants. Adipose thickness directly under the IT did not differ by diagnosis, nor did it have a consistent response to loading. However, the envelopment of the IT by the surrounding adipose tissue was much greater in two of the participants with spinal cord injuries. These two subjects also had the most curved skin surface as the tissue wrapped around the IT. Tissue strains around the ischium were most visible in the adipose and connective tissue. The gluteus maximus displaced and distorted upwards, posterior and lateral, away from the inferior IT. Multi-planar imaging is necessary to investigate anatomy and deformation of the buttocks. 5 out of 7 participants did not sit directly on muscle. The tissue beneath their ITs was predominantly composed of fat and connective tissue, suggesting that these tissues might be most vulnerable to injury.


Splicing mutations in the CFTR gene as therapeutic targets.
The marketing approval, about ten years ago, of the first disease modulator for patients with cystic fibrosis harboring specific CFTR genotypes (~5% of all patients) brought new hope for their treatment. To date, several therapeutic strategies have been approved and the number of CFTR mutations targeted by therapeutic agents is increasing. Although these drugs do not reverse the existing disease, they help to increase the median life expectancy. However, on the basis of their CFTR genotype, ~10% of patients presently do not qualify for any of the currently available CFTR modulator therapies, particularly patients with splicing mutations (~12% of the reported CFTR mutations). Efforts are currently made to develop therapeutic agents that target disease-causing CFTR variants that affect splicing. This highlights the need to fully identify them by scanning non-coding regions and systematically determine their functional consequences. In this review, we present some examples of CFTR alterations that affect splicing events and the different therapeutic options that are currently developed and tested for splice switching.