Impaired nuclear translocation of the glucocorticoid receptor in corticosteroid-insensitive airway smooth muscle in severe asthma.
Patients with severe asthma (SA) are less responsive to the beneficial effects of corticosteroid (CS) therapy, and relative CS insensitivity has been shown in airway smooth muscle cells (ASMC) from patients with SA. We investigated whether there was a defect in the actions of the glucocorticoid receptor (GR) underlying the ability of CS to suppress the inflammatory response in ASMC of patients with SA. ASMC from healthy subjects (n = 10) and subjects with severe (n = 8) and nonsevere asthma (N-SA; n = 8) were cultured from endobronchial biopsies. GR expression in ASMC from SA and N-SA was reduced compared with that from healthy subjects by 49% (P < 0.01). Although baseline levels of nuclear GR were similar, GR nuclear translocation induced by dexamethasone (10(-7) M) in SA was 60% of that measured in either healthy subjects or subjects with N-SA. Tumor necrosis factor (TNF)-α induced greater nuclear factor (NF)-κB (p65) mRNA expression in ASMC from subjects with SA (5.6- vs. 2.0-fold; P < 0.01), whereas baseline and TNF-α-induced nuclear translocation and dexamethasone-mediated suppression of p65 expression were similar between groups. Dexamethasone, although not modulating TNF-α-induced p65 nuclear translocation, attenuated p65 recruitment to the CCL11 promoter in the healthy and N-SA groups, but this suppressive effect was impaired in subjects with SA. Decreased GR expression with impaired nuclear translocation in ASMC, associated with reduced dexamethasone-mediated attenuation of p65 recruitment to NF-κB-dependent gene promoters, may underlie CS insensitivity of severe asthma.


Association of uric acid levels with cardiac syndrome X: A meta-analysis.
Objective: The pathogenesis of elevated uric acid (UA) levels in patients with cardiac syndrome X (CSX) is unclear, and the results presented in recent papers on UA levels in patients with CSX are controversial. Therefore, we conducted a meta- analysis to assess the relationship between UA levels and CSX. Methods: Three databases, including the Web of Science, EMBASE and PubMed, were systematically searched until January 2022. Fixed-effect and random-effects models were used to analyze the relationship between UA levels and CSX. Subgroup analysis and sensitivity analysis were also performed. Results: Six studies involving 406 CSX patients and 267 non-CSX were included. Our results showed a significant relationship between UA levels and CSX, with a pooled SMD of 0.68 (95% CI 0.37 to 1.00; p < 0.0001). We also found a close relationship between UA levels and CSX for patients ≥ 55 years old (SMD:0.70, 95% CI: 0.41 to 0.99, p < 0.00001), for patients < 55 years old (SMD: 0.68, 95% CI: 0.25 to 1.12, p =0 .002), for women ≥ 60% (SMD: 0.77, 95% CI: 0.33 to 1.14, p =0 .0004), for women < 60% (SMD: 0.61, 95% CI:0.23 to 0.98, p =0 .001), for BMI ≥ 28 Kg/m2 (SMD :0.61, 95% CI: 0.23 to 0.98, p =0 .001), for BMI < 28 Kg/m2 (SMD:0.75, 95% CI: 0.31 to 1.19, p =0 .0009), for publication years ≥ 2012 (SMD :0.69, 95% CI: 0.23 to 1.15, p = 0.003), for publication years < 2012 (SMD:0.73, 95% CI:0.41 to 1.05, p < 0.00001), and for Turkey (SMD:0.75, 95% CI:0.38 to 1.11, p <.0001). Sensitivity analysis showed that the pooled results remained consistent after removing any one study or converting the random-effects model to fixed-effects model. Conclusion: Our results indicated a strong association between high UA levels and CSX. However, more well-designed studies are needed to investigate whether early treatment of hyperuricemia can reduce the incidence of CSX.


Outer Membrane Vesicles.
Outer membrane vesicles (blebs) are produced by Escherichia coli, Salmonella, and all other gram-negative bacteria both in vitro and in vivo. Most of the research in the field has focused on the properties of vesicles derived from pathogenic bacteria and their interactions with eukaryotic cells. These data indicate that vesicles are able to contribute to pathogenesis. Thus, it appears that pathogenic gram-negative bacteria have co-opted vesicles for the dissemination of virulence determinants. However, the role of vesicle production by nonpathogenic bacteria is less obvious. This section reviews the data demonstrating the mechanistic and physiological basis of outer membrane vesicle production by bacteria. Vesiculation can be seen as a mechanism for cells to react to conditions in the surrounding environment by carrying away unnecessary components and allowing rapid modification of the outer membrane composition. In addition, vesicles can transmit biological activities distant from the originating cell. Vesicles could act to bind and deplete host immune factors at the site of infection that would otherwise attack the bacteria. Vesicles in the area surrounding the cell may also provide the cell protection inside a human or animal host. The concept of vesicles as virulence factors has received considerable attention, and they are likely to play a significant role in the pathogenesis of gram-negative bacteria. By analysis of their composition, mechanism of formation, regulation, and physiological function, progress is being made in understanding the ubiquitous nature of outer membrane vesicles produced by gram-negative bacteria.


Ureteric obstruction caused by pelvic actinomycosis.
Ureteric obstruction is a well-known complication of actinomycosis, however its management in previous case reports has been very variable and sometimes mutilating. We report a rare case presenting with ischiorectal abscess that was successfully treated by JJ stenting and penicillin.


Modeling Poliovirus Transmission in Borno and Yobe, Northeast Nigeria.
Beginning in 2013, multiple local government areas (LGAs) in Borno and Yobe in northeast Nigeria and other parts of the Lake Chad basin experienced a violent insurgency that resulted in substantial numbers of isolated and displaced people. Northeast Nigeria represents the last known reservoir country of wild poliovirus (WPV) transmission in Africa, with detection of paralytic cases caused by serotype 1 WPV in 2016 in Borno and serotype 3 WPV in late 2012. Parts of Borno and Yobe are also problematic areas for transmission of serotype 2 circulating vaccine-derived polioviruses, and they continue to face challenges associated with conflict and inadequate health services in security-compromised areas that limit both immunization and surveillance activities. We model poliovirus transmission of all three serotypes for Borno and Yobe using a deterministic differential equation-based model that includes four subpopulations to account for limitations in access to immunization services and dynamic restrictions in population mixing. We find that accessibility issues and insufficient immunization allow for prolonged poliovirus transmission and potential undetected paralytic cases, although as of the end of 2019, including responsive program activities in the modeling suggest die out of indigenous serotypes 1 and 3 WPVs prior to 2020. Specifically, recent and current efforts to access isolated populations and provide oral poliovirus vaccine continue to reduce the risks of sustained and undetected transmission, although some uncertainty remains. Continued improvement in immunization and surveillance in the isolated subpopulations should minimize these risks. Stochastic modeling can build on this analysis to characterize the implications for undetected transmission and confidence about no circulation.


Effect of dexmedetomidine on perioperative haemodynamics and early cognitive function in elderly patients undergoing laparoscopic surgery.
Laparoscopic minimally invasive surgery has been widely used in the diagnosis and treatment of gynaecological diseases. To investigate the effect of dexmedetomidine on perioperative haemodynamics and cognitive function in elderly gynaecological patients who underwent laparoscopic surgery. Clinical baseline characteristics, haemodynamic parameters, renin activity, norepinephrine level, cognitive function, pain level, and sedation were compared between the 2 groups. At T4 (10 min after extubation) and T5 (1 h after extubation), significant differences were found in systolic blood pressure, diastolic blood pressure, and heart rate between the 2 groups (p < 0.05); renin activity and norepinephrine level were much lower in the dexmedetomidine group than in the control group at T3 (10 min before extubation) and T4 (p < 0.05). One day before surgery, there were no significant differences in Mini-mental state examination (MMSE), visual analogue scale (VAS), and Ramsay scores between the 2 groups (p > 0.05), but the MMSE score 1 day after surgery and the Ramsay score at 12 h after surgery in the dexmedetomidine group were much higher than that in the control group (p < 0.05). Notably, at 2, 4, 12, 24, and 48 h after surgery, the VAS score in the dexmedetomidine group was significantly lower than that in the control group (p < 0.05). Dexmedetomidine has a better clinical effect in improving perioperative haemodynamics and early cognitive function in elderly gynaecological patients who received laparoscopic surgery.


Ultrasound Assisted Endovascular Thrombolysis in Adolescents: 2 Case Reports.
Descending iliofemoral thrombosis in children is a rare event. Anticoagulation therapy with low-molecular-weight-heparin is standard of care. However, patency cannot be achieved in all cases, increasing the risk for rethrombosis and postthrombotic syndrome. To reduce the risk of venous valve failure in adults, local catheter-directed thrombolysis is used to reopen vessels. Two adolescent girls (17 and 15 years old) presented with acute descending iliofemoral thrombosis of the left common iliac, external, and common femoral veins. Anticoagulation with enoxaparin was started until insertion of an EkoSonic Mach 4e catheter for ultrasound-assisted local thrombolysis with recombinant tissue plasminogen activator and administration of unfractionated heparin. Success was monitored by increases in D-dimer levels and ultrasound findings. After 24 hours respectively 48 hours, complete recanalization was obtained. No complication occurred except minimal local bleeding. Screening for hereditary thrombophilia revealed a heterozygous antithrombin mutation in 1 girl (ie, the 15-year-old). May-Thurner syndrome was identified in both girls, necessitating stenting of the left common iliac veins and continuation of anticoagulation therapy with enoxaparin and acetylsalicylic acid. No rethrombosis or complications occurred during the follow-up period. Ultrasound-assisted catheter-directed local thrombolysis with the EkoSonic Mach 4e system was effective in achieving immediate recanalization of the occluded veins and should be considered in children experiencing descending iliofemoral thrombosis. The fast recanalization might reduce the incidence of postthrombotic syndrome. May-Thurner syndrome is regularly found in these patients, and if present, requires stenting of the common iliac vein to avoid early reocclusion. However, long-term patency of iliac vein stenting in children remains to be examined.


Effect of meniscal release on rate of subsequent meniscal tears and owner-assessed outcome in dogs with cruciate disease treated with tibial plateau leveling osteotomy.
To determine and compare rates of meniscal tears after tibial plateau leveling osteotomy (TPLO) among 3 groups of dogs based on treatment method: arthrotomy with meniscal release (openR), arthrotomy without meniscal release (openNR), arthroscopy without meniscal release (scopeNR), and compare long term owner-assessed outcomes for the same groups. Retrospective cohort study. Stifles (n=254) of dogs that had TPLO. The three groups were compared for significant (P<.05) differences in rate of subsequent tears using a chi(2) test. Odds ratios for likelihood of subsequent meniscal tears were determined. Data for signalment, outcome, time to peak function, and time to subsequent tear were compared for significant differences using ANOVA, t-test, or rank sum test. Subsequent meniscal tears were diagnosed in 16 cases (6.3%). Of dogs with subsequent meniscal tears, 9 had openNR, 4 had openR, and 3 had scopeNR; the proportion of subsequent meniscal tears was significantly different (P=.035) among groups. Odds ratio indicated that subsequent meniscal tear was 3.8 times more likely to occur for openNR than openR or scopeNR. No significant differences among groups were noted for measures of outcome. Meniscal release did not reduce the rate of subsequent meniscal tears when compared with cases treated arthroscopically or when compared with all cases combined, but may be advantageous when meniscal pathology cannot be comprehensively assessed in the cranial cruciate deficient stifle. Meniscal release had no effects on owner-assessed outcome as determined in this study. The low rates of subsequent meniscal tears in conjunction with the relatively high and equivocal levels of owner-assessed outcome and time to peak function for all 3 treatment groups suggest that any of these surgical management strategies can be considered acceptable. We suggest that a meniscal release be performed when complete and thorough exploration of the joint and meniscus cannot be, or are not, performed.


Effect of Hopper Loading on the Formation of Alkyl Alcohols in Olive Fruits and Its Relationship with Sensory Quality Losses of Virgin Olive Oil.
The storage of olives in large hoppers is a widespread practice in oil mills, but these large volumes and their unloading can cause a physical deterioration of the olives that will affect the quality of the oil obtained. This research deals with the effect of hopper charge on the formation of alkyl alcohols in olive fruits and its relationship with the sensory quality losses of 'Arbequina' virgin olive oil. The contents of ethanol, methanol, and acetaldehyde were measured in olive samples loaded and stored for a short time in a large hopper and analyzed at three different hopper-discharging times, which are related to three different positions inside the hopper. The corresponding oil from each sampling was obtained by using ABENCOR and was evaluated by a trained tasting panel. Results showed that the ethanol content in olives increased during their storage in the hopper, while methanol and acetaldehyde contents did not show significant differences. Regarding their position in the hopper, fruits located at the bottom or on the lateral sides showed a greater deterioration. The sensory analyses showed an inverse relationship between the positive attributes of olive oils and their content of alcohols.


Mechanistic examination of Cβ-Cγ bond cleavages of tryptophan residues during dissociations of molecular peptide radical cations.
In this study, we used collision-induced dissociation (CID) to examine the gas-phase fragmentations of [G(n)W](•+) (n = 2-4) and [GXW](•+) (X = C, S, L, F, Y, Q) species. The C(β)-C(γ) bond cleavage of a C-terminal decarboxylated tryptophan residue ([M - CO(2)](•+)) can generate [M - CO(2) - 116](+), [M - CO(2) - 117](•+), and [1H-indole](•+) (m/z 117) species as possible product ions. Competition between the formation of [M - CO(2) - 116](+) and [1H-indole](•+) systems implies the existence of a proton-bound dimer formed between the indole ring and peptide backbone. Formation of such a proton-bound dimer is facile via a protonation of the tryptophan γ-carbon atom as suggested by density functional theory (DFT) calculations. DFT calculations also suggested the initially formed ion 2, the decarboxylated species that is active against C(β)-C(γ) bond cleavage, can efficiently isomerize to form a more stable π-radical isomer (ion 9) as supported by Rice-Ramsperger-Kassel-Marcus (RRKM) modeling. The C(β)-C(γ) bond cleavage of a tryptophan residue also can occur directly from peptide radical cations containing a basic residue. CID of [WG(n)R](•+) (n = 1-3) radical cations consistently resulted in predominant formation of [M - 116](+) product ions. It appears that the basic arginine residue tightly sequesters the proton and allows the charge-remote C(β)-C(γ) bond cleavage to prevail over the charge-directed one. DFT calculations predicted that the barrier for the former is 6.2 kcal mol(-1) lower than that of the latter. Furthermore, the pathway involving a salt-bridge intermediate also was accessible during such a bond cleavage event.


Hatching timing, oxygen availability, and external gill regression in the tree frog, Agalychnis callidryas.
The physiological role of the embryonic external gills in anurans is equivocal. In some species, diffusion alone is clearly sufficient to supply oxygen throughout the embryonic period. In others, morphological elaboration and environmental regulation of the external gills suggest functional importance. Since oxygen stress is a common trigger of hatching, I examined the relationships among hatching timing, oxygen stress, and external gill loss. I worked with the red-eyed tree frog, Agalychnis callidryas, a species with arboreal eggs and aquatic tadpoles in which gill regression is associated with hatching, and hatching timing affects posthatching survival with aquatic predators. Both exposure to a hypoxic gas mixture and submergence in water, a natural context in which hypoxic stress can occur, induced early hatching. Exposure to hyperoxic gas mixtures induced regression of external gills, and subsequent exposure to air induced early hatching. Prostaglandin-induced external gill regression also induced hatching, and this effect was partially ameliorated by exposure to hyperoxic gas. Together, these results suggest that external gills enhance the oxygen uptake of embryos and are necessary to extend embryonic development past the onset of hatching competence.


The effect of repeated loading and freeze-thaw cycling on immature bovine thoracic motion segment stiffness.
There is growing interest in the biomechanics of "fusionless" implant constructs used for deformity correction in the thoracic spine; however, there are questions over the comparability of in vitro biomechanical studies from different research groups due to the various methods used for specimen preparation, testing and data collection. The aim of this study was to identify the effect of two key factors on the stiffness of immature bovine thoracic spine motion segments: (1) repeated cyclic loading and (2) multiple freeze-thaw cycles, to aid in the planning and interpretation of in vitro studies. Two groups of thoracic spine motion segments from 6- to 8-week-old calves were tested in flexion/extension, right/left lateral bending and right/left axial rotation under moment control. Group A was tested with continuous repeated cyclic loading for 500 cycles with data recorded at cycles 3, 5, 10, 25, 50, 100, 200, 300, 400 and 500. Group (B) was tested after each of five freeze-thaw sequences, with data collected from the 10th load cycle in each sequence. Results of testing showed that for Group A: flexion/extension stiffness reduced significantly over the 500 load cycles (-22%; p = 0.001), but there was no significant change between the 5th and 200th load cycles. Lateral bending stiffness decreased significantly (-18%; p = 0.009) over the 500 load cycles, but there was no significant change in axial rotation stiffness (p = 0.137). Group B: there was no significant difference between mean stiffness over the five freeze-thaw sequences in flexion/extension (p = 0.813) and a near-significant reduction in mean stiffness in axial rotation (-6%; p = 0.07). However, there was a statistically significant increase in stiffness in lateral bending (+30%; p = 0.007). Study findings indicate that comparison of in vitro testing results for immature thoracic bovine spine segments between studies can be performed with up to 200 load cycles without significant changes in stiffness. However, when testing protocols require greater than 200 cycles, or when repeated freeze-thaw cycles are involved, it is important to account for the effect of cumulative load and freeze-thaw cycles on spine segment stiffness.


Effect of the GC content of DNA on the distribution of UVB-induced bipyrimidine photoproducts.
Solar UV radiation is a major mutagen that damages DNA through the formation of dimeric photoproducts between adjacent thymine and cytosine bases. A major effect of the GC content of the genome is thus anticipated, in particular in prokaryotes where this parameter significantly varies among species. We quantified the formation of UV-induced photolesions within both isolated and cellular DNA of bacteria of different GC content. First, we could unambiguously show the favored formation of cytosine-containing photoproducts with increasing GC content (from 28 to 72%) in isolated DNA. Thymine-thymine cyclobutane dimer was a minor lesion at high GC content. This trend was confirmed by an accurate and quantitative analysis of the photochemical data based on the exact dinucleotide frequencies of the studied genomes. The observation of the effect of the genome composition on the distribution of photoproducts was then confirmed in living cells, using two marine bacteria exhibiting different GC content. Because cytosine-containing photoproducts are highly mutagenic, it may be predicted that species with genomes exhibiting a high GC content are more susceptible to UV-induced mutagenesis.


Assessment of Brain Glucose Metabolism Following Cardiac Arrest by [18F]FDG Positron Emission Tomography.
Cardiac arrest (CA) patients who survived by cardiopulmonary resuscitation (CPR) can present different levels of neurological deficits ranging from minor cognitive impairments to persistent vegetative state and brain death. The pathophysiology of the resulting brain injury is poorly understood, and whether changes in post-CA brain metabolism contribute to the injury are unknown. Here we utilized [18F]fluorodeoxyglucose (FDG)-Positron emission tomography (PET) to study in vivo cerebral glucose metabolism 72 h following CA in a murine CA model. Anesthetized and ventilated adult C57BL/6 mice underwent 12-min KCl-induced CA followed by CPR. Seventy-two hours following CA, surviving mice were intraperitoneally injected with [18F]FDG (~ 186 µCi/200 µL) and imaged on Molecubes preclinical micro-PET/computed tomography (CT) imaging systems after a 30-min awake uptake period. Brain [18F]FDG uptake was determined by the VivoQuant software on fused PET/CT images with the 3D brain atlas. Upon completion of Positron emission tomography (PET) imaging, remaining [18F]FDG radioactivity in the brain, heart, and liver was determined using a gamma counter. Global increases in brain [18F]FDG uptake in post-CA mice were observed compared to shams and controls. The median standardized uptake value of [18F]FDG for CA animals was 1.79 versus sham 1.25 (p < 0.05) and control animals 0.78 (p < 0.01). This increased uptake was consistent throughout the 60-min imaging period and across all brain regions reaching statistical significance in the midbrain, pons, and medulla. Biodistribution analyses of various key organs yielded similar observations that the median [18F]FDG uptake for brain was 7.04%ID/g tissue for CA mice versus 5.537%ID/g tissue for sham animals, p < 0.05). This study has successfully applied [18F]FDG-PET/CT to measure changes in brain metabolism in a murine model of asystolic CA. Our results demonstrate increased [18F]FDG uptake in the brain 72 h following CA, suggesting increased metabolic demand in the case of severe neurological injury. Further study is warranted to determine the etiology of these changes.


Reversible Adsorption Kinetics of Near Surface Dimer Colloids.
We investigate the effect of shape on reversible adsorption kinetics using colloidal polystyrene dimers near a solid glass surface as a model system. The interaction between colloid and wall is tuned using electrostatic, depletion, and gravity forces to produce a double-well potential. The dwell time in each of the potential wells is measured from long duration particle trajectories. The height of each monomer relative to the glass surface is measured to a resolution of <20 nm by in-line holographic microscopy. The measured transition probability distributions are used in kinetic equations to describe the flux of particles to and from the surface. The dimers are compared to independent isolated monomers to determine the effects of shape on adsorption equilibria and kinetics. To elucidate these differences, we consider both mass and surface coverage and two definitions of surface coverage. The results show that dimers with single coverage produce slower adsorption, lower surface coverage, and higher mass coverage in comparison to those of monomers, while dimers with double coverage adsorb faster and result in higher surface coverage.


[Performance of endorectal echography in tumors of the rectum].
From November 1986 to December 1987, endorectal ultrasound (EUS) has been performed 57 times in a total of 54 patients. In the cancers of the low and mid rectum (n = 34), the extent of the tumoral infiltration was accurately assessed by preoperative EUS in 88% of cases. Sensitivity of tumor spread beyond the rectal wall was 0.96. Classification of lymph nodes was more hazardous. The muscular layer was intact in 5 villous adenomas. The diagnosis was difficult in the case of a huge and massively secreting tumour. In the follow-up of the patients after rectal resection(n = 11) or local excision (n = 3) for cancer, EUS allowed an accurate analysis of the suture line in 12 cases. Three submucosal recurrences were detected. The follow-up showed no recurrence in the other cases with a tumor-free suture line pattern. The literature confirms that EUS is a non invasive, efficient and inexpensive method in the preoperative staging of non stenotic rectal tumors. The problem of accuracy of lymph node staging has not been resolved. The technique shows some promise in detecting local recurrences.


Prevalence of metabolic syndrome and cardiovascular risk factors among voluntary screened middle-aged and elderly Egyptians.
Central obesity and diabetes mellitus are recorded at high percentages among Egyptians. The aim of this study is to determine the prevalence of metabolic syndrome (MetS) and cardiovascular risk factors among a group of middle-aged and elderly Egyptians. Our study included 220 middle-aged and senior Egyptians voluntary screened in an Egyptian private hospital with 800-bed capacity. Detailed medical history was obtained from all subjects, followed by clinical examination with weight and height measurement, body mass index calculation, waist hip ratio and arterial blood pressure measurement. Laboratory investigations done were complete blood picture, lipid profile and fasting blood glucose measurements. The diagnosis of MetS was based on the American Heart Association/Updated NCEP ATP III criteria. Cardiovascular risk assessment was calculated for each subject based on the Framingham/ATP III criteria. The prevalence of MetS in this study was of 55% among the whole sample, 85.6% among diabetics and 76.6% among hypertensive patients. Based on Framingham scoring system, 48.2% of the sample had moderate to high risk of developing cardiovascular disease. Odds ratio for patients with MetS for developing cardiovascular disease in the next 10 years was 2.8 (95% confidence interval: 1.6-4.8). The high prevalence of MetS among middle-aged and elderly Egyptians with the documented high prevalence of chronic diseases in Egypt calls for a nationwide screening program to detect MetS and tackle preventive strategies to face the epidemic of obesity and outcomes of MetS, particularly cardiovascular diseases.


Development and characterization of a monoclonal antibody which distinguishes the beta-subunit of human chorionic gonadotropin (beta hCG) in the presence of the hCG.
A monoclonal antibody specific for the beta-subunit of human chorionic gonadotropin (beta hCG) has been developed which has a cross reactivity of 0.23% with intact hCT. There was no cross reactivity with other glycoprotein pituitary hormones in the range in which they might be expected to be present in serum. The beta-subunits of hFSH, hTSH and hLH did not inhibit binding of [125I]iodo-beta hCG to the antibody at up to 250 ng/tube. The dissociation constant (Kd) with [125I]iodo-beta hCG was 3.3 x 10(-10), and the reaction had reached equilibrium within 1 h.


The effects of dose staggering on metabolic drug-drug interactions.
To investigate the effect of dose staggering on metabolic drug-drug interactions (MDDI). Using Matlab, anatomical, physiological and biochemical data relating to human pharmacokinetics were integrated to create a representative virtual healthy subject relevant to in vivo studies. The effects of dose staggering on AUC and C(max) were investigated under various scenarios with respect to pharmacokinetic characteristics of the inhibitor and substrate drugs (e.g. hepatic extraction ratio). Specific cases were also simulated where MDDI had been studied experimentally for combinations of drugs (budesonide and ketoconazole; triazolam and itraconazole). The decrease in the magnitude of the inhibitory effect of the 'perpetrator' drug (inhibitor) on the 'victim' drug (substrate) as a result of 'dose staggering' was greater when the 'perpetrator' was given after the 'victim'. There was reasonable agreement between the predicted extent of the interactions and the observed in vivo data (mean prediction errors of 25 and -14% for AUC and C(max) values, respectively (n=7)). The impact of dose staggering was minimal during continuous dosage of inhibitors with long elimination half-lives (e.g. itraconazole, >20 h). Clinical trial simulations using physiological information may provide useful guidelines for optimal dose staggering when poly-pharmacy is inevitable.


[The clinical characteristics of 70 cases of fungemia].
To evaluate the etiological and clinical characteristics of fungemia in Peking Union Medical College Hospital. Microbial and clinical information of patients with fungemia consulted in Peking Union Medical College Hospital during 2008 to 2010 were retrospectively analyzed. A total of 70 patients were diagnosed with fungemia, and 100% of them had underlying diseases or potential risk factors. Of them, 40 (57.1%) patients were monomicrobial fungemia, and the other 30 (42.9%) patients with positive blood cultures were caused by at least two different microbes during hospitalization. Among 122 strains of microbes isolated from blood cultures, 72 were fungi and 50 were bacteria. Among the isolated fungi, 61 (84.7%) were Candida species, 31 (50.8%) were Candida albicans and 30 (49.2%) were non-albicans. According to the colonization or infected sites other than blood of the isolated fungi, 35 cases (50.0%) were primary fungemia; 18 (25.7%) were colonizing at lower respiratory tract simultaneously; 10 (14.3%) caused central-line related fungemia; 3(4.3%) were secondary to intra-abdominal fungal infection; and another 4 (5.7%) isolates had multiple colonization sites. During hospitalization, 37 cases died with a crude mortality rate of 52.9%, and 22 (32.9%) died of fungemia itself. In single factor analysis, ICU hospitalization (χ(2) = 15.136, P < 0.001), operation history within 30 days (χ(2) = 3.540, P = 0.060) and invasive mechanical ventilation (χ(2) = 4.450, P = 0.035) were related to crude mortality. Bacteremia during hospitalization (χ(2) = 5.657, P = 0.017), circulatory underlying diseases (χ(2) = 3.399, P = 0.065) and ICU treatment (χ(2) = 4.955, P = 0.026) increased attributable mortality. In the multivariate analysis, ICU history increased mortality during hospitalization, however, the operation history within 30 days was independently irrelevant to crude mortality during hospitalization. ICU history and bacteremia during hospitalization were independently correlated to attribution mortality of the patients with fungemia. Fungemia, usually accompanied with bacteremia, occurs often in the patients with underlying diseases. Patients with fungemia have poor prognosis and more than 50% patients die. ICU history increases the risk both to crude and attributable mortality. The patients with fungemia who had polymicrobial bloodstream infection have a higher attribution mortality. Operation history within 30 days is independently negatively correlated to attributable mortality.


A triple-blind, placebo-controlled randomised trial of the ilioinguinal-transversus abdominis plane (I-TAP) nerve block for elective caesarean section.
This study investigated the efficacy of a new ilioinguinal-transversus abdominis plane block when used as a component of multimodal analgesia. We conducted a prospective, triple-blind, placebo-controlled randomised study of 100 women undergoing elective caesarean section. All women had spinal anaesthesia with hyperbaric bupivacaine, 15 μg fentanyl and 150 μg morphine, as well as 100 mg diclofenac and 1.5 g paracetamol rectally. Women were randomly allocated to receive the ilioinguinal-transversus abdominis plane block or a sham block at the end of surgery. The primary outcome was the difference in fentanyl patient-controlled analgesia dose at 24 h. Secondary outcomes included postoperative pain scores, adverse effects and maternal satisfaction. The cumulative mean (95%CI) fentanyl dose at 24 h was 71.9 (55.6-92.7) μg in the ilioinguinal-transversus abdominis group compared with 179.1 (138.5-231.4) μg in the control group (p < 0.001). Visual analogue scale pain scores averaged across time-points were 1.9 (1.5-2.3) mm vs. 5.0 (4.3-5.9) mm (p = 0.006) at rest, and 4.7 (4.1-5.5) mm vs. 11.3 (9.9-13.0) mm (p = 0.001) on movement, respectively. Post-hoc analysis showed that the ilioinguinal-transversus abdominis group was less likely to use ≥ 1000 μg fentanyl compared with the control group (2% vs. 16%; p = 0.016). There were no differences in opioid-related side-effects or maternal satisfaction with analgesia. The addition of the ilioinguinal-transversus abdominis plane block provides superior analgesia to our usual multimodal analgesic regimen.


A comparison of reproductive characteristics of boars generated by somatic cell nuclear transfer to highly related conventionally produced boars.
This study compares the reproductive performance of boars produced by somatic cell nuclear transfer versus conventional breeding. Two different genotypes were selected for comparison: terminal cross line 1 (TX1) and terminal cross line 2 (TX2). The boars selected for comparison from TX1 were three cloned boars, produced by somatic cell nuclear transfer and the conventionally produced progenitor of the clones. The boars selected for comparison from TX2 were a cloned boar produced by somatic cell nuclear transfer and two conventionally produced half sibling boars that were offspring of the progenitor of the clone. Semen from each boar was collected, extended, evaluated and shipped offsite. Upon arrival, the semen was reevaluated and utilized for artificial insemination of 89 commercial gilts, at least 12 gilts per boar, producing 625 piglets. Pregnancy rates were determined at day 30 and 110 of gestation; and farrowing rate and gestation length were recorded. Differences were observed in some of the semen characteristics analyzed with the clones usually possessing superior semen quality to the control, this likely being a result of age differences amongst the clones and controls. Additionally no differences were noted between the clones and controls (progenitor) or between individual boars within genetic line for pregnancy rates, gestation length or any of the litter parameters examined between the clones and controls. These data further support previous reports with limited numbers that the reproductive capabilities of cloned boars are equal to that of conventionally produced boars.


Abundant nucleostemin expression supports the undifferentiated properties of germ cell tumors.
Nucleostemin (NS) is a nucleolar GTP-binding protein that is involved in ribosomal biogenesis and protection of telomeres. We investigated the expression of NS in human germ cell tumors and its function in a mouse germ cell tumor model. NS was abundantly expressed in undifferentiated, but not differentiated, types of human testicular germ cell tumors. NS was expressed concomitantly with OCT3/4, a critical regulator of the undifferentiated status of pluripotent stem cells in primordial germ cells and embryonal carcinomas. To investigate the roles of NS in tumor growth in vivo, we used a mouse teratoma model. Analysis of teratomas derived from embryonic stem cells in which the NS promoter drives GFP expression showed that cells highly expressing NS were actively proliferating and exhibited the characteristics of tumor-initiating cells, including the ability to initiate and propagate tumor cells in vivo. NS-expressing cells exhibited higher levels of GTP than non-NS-expressing cells. Because NS protein is stabilized by intracellular GTP, metabolic changes may contribute to abundant NS expression in the undifferentiated cells. OCT3/4 deficiency in teratomas led to loss of NS expression, resulting in growth retardation. Finally, we found that teratomas deficient in NS lost their undifferentiated characteristics, resulting in defective tumor proliferation. These data indicate that abundant expression of NS supports the undifferentiated properties of germ cell tumors.


GMOMETHODS: the European Union database of reference methods for GMO analysis.
In order to provide reliable and harmonized information on methods for GMO (genetically modified organism) analysis we have published a database called "GMOMETHODS" that supplies information on PCR assays validated according to the principles and requirements of ISO 5725 and/or the International Union of Pure and Applied Chemistry protocol. In addition, the database contains methods that have been verified by the European Union Reference Laboratory for Genetically Modified Food and Feed in the context of compliance with an European Union legislative act. The web application provides search capabilities to retrieve primers and probes sequence information on the available methods. It further supplies core data required by analytical labs to carry out GM tests and comprises information on the applied reference material and plasmid standards. The GMOMETHODS database currently contains 118 different PCR methods allowing identification of 51 single GM events and 18 taxon-specific genes in a sample. It also provides screening assays for detection of eight different genetic elements commonly used for the development of GMOs. The application is referred to by the Biosafety Clearing House, a global mechanism set up by the Cartagena Protocol on Biosafety to facilitate the exchange of information on Living Modified Organisms. The publication of the GMOMETHODS database can be considered an important step toward worldwide standardization and harmonization in GMO analysis.


Immune Microenvironment in Microsatellite-Instable Endometrial Cancers: Hereditary or Sporadic Origin Matters.
Purpose: Recent studies show that colorectal tumors with high microsatellite instability (MSI-H) have increased immunogenicity and response to immunotherapy compared with microsatellite-stable (MSS) tumors. It is not yet clear whether MSI-H endometrial cancer may also benefit from these therapies. It is also unknown whether immune response is equivalent in MSI-H endometrial cancer with sporadic or inherited Lynch syndrome origins.Experimental Design: Multiplexed fluorescent IHC was used to compare matched MSI-H (n = 60) and MSS (n = 96) endometrial cancer specimens by evaluating immune cell populations in tumor and stroma compartments. Sporadic MSI-H and Lynch syndrome-associated (LS) MSI-H endometrial cancers were also directly compared.Results: Increased immune cells were present in stroma of MSI-H endometrial cancer compared with MSS, including granzyme B+ cells, activated CTLs (CD8+granzyme B+), and PD-L1+ cells. Granzyme B+ cells and activated CTLs were also increased in the tumor compartment of MSI-H endometrial cancers. Comparing sporadic and LS MSI-H endometrial cancer showed distinct differences in immune cell populations, indicating that mechanisms underlying microsatellite instability alter immune response. Specifically, LS MSI-H endometrial cancer showed increased CD8+ cells and activated CTLs in stroma, with reduced macrophages in stroma and tumor compared with sporadic MSI-H. Sporadic MSI-H had increased PD-L1+ macrophages in stroma and tumor compared with LS MSI-H endometrial cancer.Conclusions: MSI-H endometrial cancer has increased immune cell infiltration compared with MSS endometrial cancer and the hereditary or sporadic origin of microsatellite instability impacts immune response. Clinical trials to determine the role of immunotherapy in patients with MSI-H endometrial cancer must evaluate Lynch syndrome-related and sporadic MSI-H tumors separately. Clin Cancer Res; 23(15); 4473-81. ©2017 AACR.


Analog computing by Brewster effect.
Optical computing has emerged as a promising candidate for real-time and parallel continuous data processing. Motivated by recent progresses in metamaterial-based analog computing [Science343, 160 (2014)SCIEAS0036-807510.1126/science.1242818], we theoretically investigate the realization of two-dimensional complex mathematical operations using rotated configurations, recently reported in [Opt. Lett.39, 1278 (2014)OPLEDP0146-959210.1364/OL.39.001278]. Breaking the reflection symmetry, such configurations could realize both even and odd Green's functions associated with spatial operators. Based on such an appealing theory and by using the Brewster effect, we demonstrate realization of a first-order differentiator. Such an efficient wave-based computation method not only circumvents the major potential drawbacks of metamaterials, but also offers the most compact possible device compared to conventional bulky lens-based optical signal and data processors.


Successful treatment of Rosai-Dorfman disease using in situ photoimmunotherapy.
Rosai-Dorfman disease is difficult to cure. In situ photoimmunotherapy combines local photothermal therapy with immunoadjuvant. In the present case report, a 39-year-old Chinese man with Rosai-Dorfman disease lesions below the left nostril and left preauricular region was treated with in situ photoimmunotherapy. The patient was treated with daily application of topical imiquimod (5%) and laser irradiations every 2 weeks for 8 weeks. After three cycles of treatment, the lesions improved markedly without adverse effects. Our results showed that in situ photoimmunotherapy can be used as an effective treatment for Rosai-Dorfman disease.


Antifungal activity of isothiocyanates and related compounds. 3. Derivatives of biphenyl, stilbene, azobenzene, and several polycondensed aromatic hydrocarbons.
This paper presents the results of a study on the antifungal activity of isothiocyanates-derivatives of biphenyl (group "A"), of stilbene ("B"), of azobenzene and benzeneazonaphthalene ("C"), of naphthalene ("D"), and of further polycondensed aromatic hydrocarbons ("E"). From a total of 48 investigated compounds, antifungal activity was observed only in A and D group compounds. B, C, and E group derivatives are extremely insoluble in water, and the molecules are very large; as a result, they probably cannot pass into spores or mycelium of fungi. Thus, the -NCS group cannot manifest its reactivity.


Superionicity and polymorphism in calcium fluoride at high pressure.
We present a combined experimental and computational first-principles study of the superionic and structural properties of CaF_{2} at high P-T conditions. We observe an anomalous superionic behavior in the low-P fluorite phase that consists of a decrease of the normal → superionic critical temperature with compression. This unexpected effect can be explained in terms of a P-induced softening of a zone-boundary X phonon that involves exclusively fluorine displacements. Also we find that superionic conductivity is absent in the high-P cotunnite phase. Instead, superionicity develops in a new low-symmetry high-T phase that we identify as monoclinic (space group P2_{1}/c). We discuss the possibility of observing these intriguing phenomena in related isomorphic materials.


A new variant of late-onset myophosphorylase deficiency.
McArdle disease classically presents in childhood or adolescence. Rarely does it become symptomatic for the first time in late adulthood, with the onset of progressive muscle wasting and weakness. Our patient is unusual in that despite a life of physical vigor, she developed immobilizing cramps, stiffness, and muscle swelling abruptly at age 60. She had no previous symptoms of muscle disease. The diagnosis was indicated by the ischemic forearm test, which produced muscle contracture and no rise in venous lactate levels, and confirmed by histochemical, electrophoretic, and biochemical studies that showed complete absence of myophosphorylase. This case defines a new variant of the late-onset type and raises important questions about compensatory mechanisms, inheritance patterns, and etiological factors in myophosphorylase deficiency.


Binding of pp170 to microtubules is regulated by phosphorylation.
We have used a monoclonal antibody affinity column to purify from HeLa cells a protein of molecular weight 170,000 (designated pp170) which we previously identified as a nucleotide-sensitive microtubule-binding protein (Rickard, J. E., and Kreis, T. E. (1990) J. Cell Biol. 110, 1623-1633). We show here that the affinity-purified pp170 binds directly to taxol-polymerized tubulin. This association is not affected directly by MgATP. Addition of MgATP can, however, inhibit binding of pp170 to microtubules in the presence of microtubule-binding proteins from HeLa cells. This effect of MgATP correlates with phosphorylation of pp170 by a microtubule-associated kinase. Potato acid phosphatase dephosphorylates the pp170 and restores the ability of pp170 to bind to microtubules. Furthermore, binding of pp170 to microtubules in a high speed supernatant extract is inhibited by the phosphatase inhibitor okadaic acid, consistent with an inhibitory effect of pp170 phosphorylation on microtubule binding. In vivo, pp170 is phosphorylated on serine residues, with a half-life for the phosphate groups of approximately 2 h. Depolymerization of microtubules with nocodazole abolishes incorporation of 32P into the protein, apparently by increasing the rate of its dephosphorylation. Stabilization of microtubules with taxol reduces the rate of 32P incorporation into pp170 by approximately 50%, but has no significant effect on phosphate loss. These data establish that pp170 is a microtubule-binding protein, and that the microtubule interaction is inhibited by phosphorylation of pp170. The sensitivity of the in vivo phosphorylation state of pp170 to microtubule-active drugs suggests that this posttranslational modification may be an important regulator of the interaction of pp170 with microtubules in cells.


[The basis of treatment with recombinant human erythropoietin].
A line of transgenic mice overexpressing erythropoietin was created. These mice retained their capacity to reduce their gastro-intestinal absorption of iron and to regulate the changes in their iron metabolism and they could serve as a model for the in vivo study of iron homeostasis and erythropoiesis. NEW INDICATIONS FOR RECOMBINANT HUMAN ERYTHROPOIETIN: After chronic terminal kidney failure, the treatment of chronic dialysed kidney failure patients and patients treated with azathioprine or patients having undergone surgery and requiring transfusion, other indications have been proposed. Such as anaemia in children following inadequate production of endogenous erythropoietin and/or direct inhibition of the erythroid cell line in the bone marrow or anaemia during pregnancy and, since the Sixties, anaemia during cancer. TO ASSESS THE PHYSIOPATHOLOGY OF ANEMIA:In anaemic patients suffering from a malignant blood disease, it would be useful to calculate the relationship between the predicted and observed rates of erythropoietin as well as the transferin serum receptors.


Tracking bacterial DNA patterns in septic progression using 16s rRNA gene amplicon sequencing analysis.
Bloodstream infections remain prevalent in intensive care units, leading to a public health challenge worldwide. Routine diagnosis is mainly based on blood culture, but the technique is limited by its time-consuming process and relatively low sensitivity. Emerging molecular diagnostic tools, such as 16S metagenomics, have been developed for detecting bacteria in the blood samples of septic patients. Using a collection of 168 blood samples from 96 septic patients, 16S metagenomics method followed by bioinformatics were applied to study bacterial alterations during the pathogenesis of sepsis. Significant taxonomic variations were found between the two survival groups at different therapeutic time points through sequential 16S metagenomics research. The results on the third day during the treatment course were notably distinct among the studied groups. 16S metagenomics approach can bring novel genetic insight about microbiological fluctuations during septic progression, which may be utilized as a complementary prognostic application. Further etiologic and pathophysiologic explorations are needed to fully explain the linkage between clinical outcomes and genetic changes.


Scrofulous swelling of the bosom masquerading as cancer.
Tuberculosis of breast is very rare with an incidence of 0.1-0.5%. It can be primary or secondary. Except in patients presenting with sinuses, it is a challenge to diagnose it. A 40 year old premenopausal lady presented with breast lump increasing in size for 3 months. Mammogram showed a lesion suspicious of malignancy and trucut biopsy showed necrotic material only. Intraoperatively there was caseous necrosis and the tract from breast was extending to rib. It is a rare case with few case reports been reported where a rib tuberculosis presents as a breast lump rather than retromammary abscess.


Quantitative determination of the NMDA antagonist cis-4-phosphonomethyl-2-piperidine carboxylic acid (CGS 19755) in human plasma using capillary gas chromatography/mass spectrometry.
An analytical method has been developed and validated for the quantitative determination of the N-methyl-D-aspartate (NMDA) antagonist cis-4-phosphonomethyl-2-piperidine carboxylic acid (CGS 19755) in human plasma. It is a member of a new class of compounds with the potential to be neuroprotective and attenuate neuronal damage resulting from brain trauma caused by stroke and head trauma. The method is based on gas chromatography/mass spectrometry and uses stable-isotope labeled CGS 19755 as the internal standard. Samples (1 ml) were first acidified (pH 2), then extracted using a solid-phase aminopropyl ion exchange column. The drug was eluted with NH4OH and evaporated until dry. Extracts were derivatized with a mixture of pentafluoropropionic anhydride and pentafluoropropanol, and analyzed by gas chromatography/mass spectrometry. Separation was accomplished on a DB-225 capillary column (15 m x 0.32 mm) with a 0.25 micron film thickness. Mass spectrometry was carried out under negative ion ammonia chemical ionization conditions with selected ion monitoring at m/z 760 and 764 for derivatized CGS 19755 and the internal standard, respectively. Specificity was shown by the lack of interfering peaks at the retention time of CGS 19755 and internal standard. Recovery and reproducibility assessments show good accuracy, precision and linearity over the validated concentration range of 2-5000 ng ml-1.


Megacities and climate change - A brief overview.
Cities have developed into the hotspots of human economic activity. From the appearance of the first cities in the Neolithic to 21st century metropolis their impact on the environment has always been apparent. With more people living in cities than in rural environments now it becomes crucial to understand these environmental impacts. With the immergence of megacities in the 20th century and their continued growth in both, population and economic power, the environmental impact has reached the global scale. In this paper we examine megacity impacts on atmospheric composition and climate. We present basic concepts, discuss various definitions of footprints, summarize research on megacity impacts and assess the impact of megacity emissions on air quality and on the climate at the regional to global scale. The intention and ambition of this paper is to give a comprehensive but brief overview of the science with regard to megacities and the environment.


Structural Basis for the Decoding Mechanism.
The bacterial ribosome is a complex macromolecular machine that deciphers the genetic code with remarkable fidelity. During the elongation phase of protein synthesis, the ribosome selects aminoacyl-tRNAs as dictated by the canonical base pairing between the anticodon of the tRNA and the codon of the messenger RNA. The ribosome's participation in tRNA selection is active rather than passive, using conformational changes of conserved bases of 16S rRNA to directly monitor the geometry of codon-anticodon base pairing. The tRNA selection process is divided into an initial selection step and a subsequent proofreading step, with the utilization of two sequential steps increasing the discriminating power of the ribosome far beyond that which could be achieved based on the thermodynamics of codon-anticodon base pairing stability. The accuracy of decoding is impaired by a number of antibiotics and can be either increased or decreased by various mutations in either subunit of the ribosome, in elongation factor Tu, and in tRNA. In this chapter we will review our current understanding of various forces that determine the accuracy of decoding by the bacterial ribosome.


Endocrine characteristics of polycystic ovary syndrome (PCOS).
Polycystic ovary syndrome (PCOS) is probably the most prevalent endocrinopathy in women and the most common cause of anovulatory infertility. Patients with PCOS have clinical and biochemical features consistent with the ultrasound diagnosis and they are likely to face the problems of hyperandrogenism, subfertility and recurrent miscarriage. The aim of the present review is to summarize our present knowledge on the hormonal background of this very prevalent syndrome and to give some clinical examples how the present knowledge can be applied to treat PCOS patients according to their current problem, such as menstrual cycle disorder, hirsutism, infertility or to prevent late consequences as diabetes mellitus. The etiology and pathogenesis of PCOS is still a matter of controversies, but it is apparent that inappropriate gonadotropin secretion, obesity, hyperinsulinism and insulin resistance are the major determining factors in the development of ovarian hyperandrogenism an chronic anovulation. Reversal of insulin resistance in PCOS constitutes the fundamental goal in the management of hyperandrogenic anovulatory infertility and in the prevention of long-term consequences. The value of the insulin sensitizer metformin therapy awaits further evaluation and it should be integrated in the spectrum of therapeutical options that include the discussed surgical methods and GnRH analogues as well.


Characterization and taxonomic placement of Rhizoctonia-like endophytes from orchid roots.
Twenty-one Rhizoctonia-like fungal strains were isolated from the roots of four terrestrial orchid species from various locations in Hong Kong. The cultural morphology, nuclear number of the hyphal cell, pore ultrastructure, and RAPD and CAPS analyses of rDNA fragments revealed that most of these isolates were associated with the genera Ceratorhiza and Epulorhiza. RAPD analysis showed the presence of genetic diversity between the isolates from different hosts and locations. The compatibility between a selection of these Ceratorhiza and Epulorhiza isolates and 14 orchid species was determined using a symbiotic germination method. The germination and development of three orchid species, Arundina chinensis, Spathoglottis pubescens, and Spiranthes hongkongensis, were strongly stimulated by the Epulorhiza isolates. Habenaria dentata was found to form symbionts successfully with a Ceratorhiza isolate.


Secretory processing of amyloid precursor protein is inhibited by increase in cellular cholesterol content.
Plasma-membrane composition plays a crucial role in most of the cellular functions that depend on membrane processes. In virtually all cell types the proteolytic processing of Alzheimer amyloid precursor protein (APP) to generate soluble APP (sAPP) is believed to occur at the plasma membrane or in its immediate proximity. Alteration of this metabolic pathway has been linked to the pathogenesis of Alzheimer's disease. We analysed the effect of membrane cholesterol enrichment on APP metabolism. Incubation of COS cells with increasing concentrations of non-esterified cholesterol carried by rabbit beta-very low-density lipoprotein caused a dose-dependent inhibition of sAPP release: 70% inhibition with 10 microg/ml non-esterified cholesterol. A less pronounced inhibitory effect was observed on treatment with human low-density lipoprotein. Inhibition of sAPP release was independent of receptor-mediated lipoprotein metabolism since simultaneous treatment with chloroquine did not modify the effect of lipoprotein treatment. In addition, treatment with cholesterol dissolved in either ethanol or methyl-beta-cyclodextrin elicited the same effect. Excess non-esterified cholesterol did not cause cell toxicity. Cell cholesterol mass inversely correlated with sAPP release. Progesterone, which inhibits shuttling of non-esterified cholesterol between the plasma membrane and intracellular pools, had no effect on the inhibition of sAPP release from cholesterol-loaded cells, providing indirect evidence that cholesterol may act at the plasma membrane.


Platelet Detection Based on Improved YOLO_v3.
Platelet detection and counting play a greatly significant role in medical field, especially in routine blood tests which can be used to judge blood status and diagnose related diseases. Therefore, platelet detection is valuable for diagnosing related blood diseases such as liver-related diseases. Blood analyzers and visual microscope counting were widely used for platelet detection, but the experimental procedure took nearly 20 minutes and can only be performed by a professional doctor. In recent years, technological breakthroughs in artificial intelligence have made it possible to detect red blood cells through deep learning methods. However, due to the inaccessibility of platelet datasets and the small size of platelets, deep learning-based platelet detection studies are almost nonexistent. In this paper, we carried out experiments for platelet detection based on commonly used object detection models, such as Single Shot Multibox Detector (SSD), RetinaNet, Faster_rcnn, and You Only Look Once_v3 (YOLO_v3). Compared with the other three models, YOLO_v3 can detect platelets more effectively. And we proposed three ideas for improvement based on YOLO_v3. Our study demonstrated that YOLO_v3 can be adopted for platelet detection accurately and in real time. We also implemented YOLO_v3 with multiscale fusion, YOLO_v3 with anchor box clustering, and YOLO_v3 with match parameter on our self-created dataset and, respectively, achieved 1.8% higher average precision (AP), 2.38% higher AP, and 2.05% higher AP than YOLO_v3. The comprehensive experiments revealed that YOLO_v3 with the improved ideas performs better in platelet detection than YOLO_v3.


Nanoparticles increase human bronchial epithelial cell susceptibility to respiratory syncytial virus infection via nerve growth factor-induced autophagy.
Cytotoxic and neuroinflammatory effects of TiO2 nanoparticles (TiO2-NP) in human airways are mediated by nerve growth factor (NGF), which is also implicated in the pathophysiology of respiratory syncytial virus (RSV) infection. We tested the hypothesis that exposure to TiO2-NP results in increased susceptibility to RSV infection and exacerbation of airway inflammation via NGF-mediated induction of autophagy in lower respiratory tract cells. Human primary bronchial epithelial cells were exposed to TiO2-NP for 24 h prior to infection with recombinant red RSV (rrRSV). Expression of NGF and its TrkA and p75NTR receptors was measured by real-time PCR and fluorescence-activated cell sorting (FACS). Autophagy was assessed by beclin-1 expression analysis. Cell death was studied by FACS after annexin V/propidium iodide staining. rrRSV infection efficiency more than doubled in human bronchial cells pre-exposed to TiO2-NP compared to controls. NGF and its TrkA receptor were upregulated in RSV-infected bronchial cells pre-exposed to TiO2-NP compared to controls exposed to either rrRSV or TiO2-NP alone. Silencing NGF gene expression with siRNA significantly inhibited rrRSV infection. rrRSV-infected cells pre-exposed to TiO2-NP also showed increase in necrotic cell death and reduction in apoptosis, together with 4.3-fold increase in expression of the early autophagosomal gene beclin-1. Pharmacological inhibition of beclin-1 by wortmannin resulted in increased apoptotic rate along with lower viral load. This study shows that TiO2-NP exposure enhances the infectivity of RSV in human bronchial epithelial cells by upregulating the NGF/TrkA axis. The mechanism of this interaction involves induction of autophagy promoting viral replication and necrotic cell death.


Apoptosis, inflammatory response and parasite load in skin of Leishmania (Leishmania) chagasi naturally infected dogs: a histomorphometric analysis.
The skin has an important role in infection by Leishmania chagasi. Apoptosis modulates the inflammatory response acting distinctively either on the progression or regression of the lesions. The parasites interact with multiple regulatory systems inducing apoptosis in host cells, during cell invasion, stabilization and multiplication of pathogens. In this context, the aim of this study was to evaluate cell death within the inflammatory infiltrates, and to correlate these results with parasite load and clinical features of dogs naturally infected with L. chagasi. Fragments of skin pinnas (8 symptomatic+8 asymptomatic+6 negative controls) were used to characterize and measure the inflammatory response, parasite load and apoptosis. Diagnosis of canine leishmaniasis was confirmed by the detection of anti-Leishmania antibodies by IFA and ELISA in serum, direct visualization of the parasite and culture in spleen, liver, pinna, bone marrow and lymph nodes, and PCR (pinna). Histomorphometry was performed with images obtained from 20 representative histological fields in a light microscope. Ultra-thin sections were mounted over a 300 mesh grids, contrasted with 2% uranyl acetate and lead citrate and examined under a Transmission Electronic Microscopy. Amastigotes were only found in the skin of symptomatic animals (31.94 ± 18.81). The number of foci and cellularity of the inflammatory infiltrates in symptomatic dogs were higher than in other groups and in asymptomatics were higher than in controls (p<0.05; Tukey). The average area, perimeter and extreme diameters of the inflammatory infiltrates obtained in symptomatic dogs were higher than in controls (p<0.05; Tukey). The apoptotic index was higher in symptomatic than in other groups and there was no difference between asymptomatics and controls (p<0.05; Tukey). Ultrastructurally, apoptotic cells were shrunken, with condensed nuclear chromatin and cytoplasm. Condensed nuclei were frequently fragmented. Internucleosomal DNA fragmentation occurred only in symptomatic cases. Amastigotes were observed within neutrophils and macrophages. Apoptosis is directly related to parasite load, intensity of inflammatory response and clinical manifestations in L. chagasi naturally infected dogs.


Sleep and Respiration in 100 Healthy Caucasian Sleepers--A Polysomnographic Study According to American Academy of Sleep Medicine Standards.
Despite differences between American Academy of Sleep Medicine (AASM) and Rechtschaffen and Kales scoring criteria, normative values following the current AASM criteria are lacking. We investigated sleep and respiratory variables in healthy adults over the lifespan, and established polysomnographic normative values according to current standards. Prospective polysomnographic investigation. Academic referral hospital sleep laboratory. One hundred healthy sleepers aged 19-77 y were selected from a representative population sample by a two-step screening. N/A. All subjects underwent one full-night polysomnography. Sleep and arousals were scored according to AASM standards. Respiration was scored according to AASM 2007 and 2012 criteria in order to compare both methods. Percentile curves showed age-related differences in sleep architecture: a decrease was found for sleep efficiency [≤ 30 y: 87.0 (71.9-94.1)% versus > 60 y: 79.7 (44.5-90.9)%], total sleep time [≤ 30 y: 413.5 (345.6-451.9) min versus > 60 y: 378.3 (216.0-440.0) min], the percentages of N3 [≤ 30 y 20.7 (15.2-37.5)% versus > 60 y: 14.9 (2.4-35.6)%] and rapid eye movement sleep [≤ 30 y 15.5 (7.5-23.6)% versus. > 60 y: 10.3 (1.9-21.9)%], whereas the percentage of wake time after sleep onset increased with age [≤ 30 y 6.0 (1.9-22.8)% versus > 60 y: 15.2 (6.3-48.7)%]. The apnea-hypopnea index (AHI) was higher when applying the AASM 2012 criteria [AHI AASM 2007 0.7 (0.0-21.5)/h versus 2012: 1.7 (0.0-25)/h; P < 0.001]. Eight percent of subjects had an AHI > 15/h. This study provides normative data on sleep macrostructure, microstructure, and respiration in adults following AASM standards. Furthermore, we demonstrated that respiration scoring according to AASM 2012 results in higher AHIs, and challenge the use of age-independent respiratory cutoff values.


The Effects of Acute Copper and Ammonia Challenges on Ammonia and Urea Excretion by the Blue Crab Callinectes sapidus.
Copper (Cu) is a persistent environmental contaminant that elicits several physiological disturbances in aquatic organisms, including a disruption in ammonia regulation. We hypothesized that exposure to Cu in a model crustacean (blue crab, Callinectes sapidus) acclimated to brackish water (2 ppt) would lead to hyperammonemia by stimulating an increase in ammonia production and/or by inhibiting ammonia excretion. We further hypothesized that urea production would represent an ammonia detoxification strategy in response to Cu. In a pilot experiment, exposure to 0, 100, and 200 µg/L Cu for 6 h caused significant concentration-dependent increases in ammonia excretion (J amm). Based on these results, an acute 24-h 100 µg/L Cu exposure was conducted and this similarly caused an overall stimulation of J amm during the 24-h period, indicative of an increase in ammonia production. Terminal haemolymph total ammonia content (T amm) was unchanged, suggesting that while ammonia production was increased, there was no inhibition of the excretion mechanism. In support of our second hypothesis, urea excretion (J urea) increased in response to Cu exposure; haemolymph [urea] was unaffected. This suggested that urea production also was increased. To further test the hypothesis that J urea increased to prevent hyperammonemia during Cu exposure, crabs were exposed to high environmental ammonia (HEA; 2.5 mmol/L NH4HCO3) for 12 h in a separate experiment. This led to a fourfold increase in haemolymph T amm, whereas J urea increased only transiently and haemolymph [urea] was unchanged, indicating that urea production likely does not contribute to the attenuation of hyperammonemia in blue crabs. Overall, Cu exposure in blue crabs led to increased ammonia and urea production, which were both eliminated by excretion. These results may have important implications in aquaculture systems where crabs may be exposed to elevated Cu and/or ammonia.


Infection-related spontaneous atlantoaxial dislocation in an adult. Case report.
This paper reports the third described case of infection-related atlantoaxial subluxation in an adult. Like most of the similar cases seen in the pediatric literature, this case was associated with a parapharyngeal beta-hemolytic streptococcal abscess. Based upon this experience, the authors advocate intravenous antibiotic therapy and 1) immediate reduction followed by application of a halo brace; 2) immobilization in a halo brace for at least 3 months; and 3) a C1-2 wiring and fusion procedure for patients who fail this trial of conservative therapy.


Histochemical quantitation of gamma-glutamyl transferase in human leukemia cell lines.
The enzyme gamma-glutamyl transferase (gamma-GT) is involved in many biochemical systems, including the signal transduction of hematopoietic growth factors. Standard colorimetric gamma-GT assays require larger cell numbers than may be obtainable in many cases, such as with highly purified stem-cell populations. To study gamma-GT expression in limited populations, we used a histochemical stain to analyze gamma-GT semiquantitatively in cells of hematopoietic origin. Several human leukemic cell lines, including one with inducible increases in gamma-GT, were stained for gamma-GT and graded 0 through 4+ for the amount of positive granules. The gamma-GT activity demonstrated by this stain was found to be directly proportional to the gamma-GT activity obtained with a colorimetric assay and could be used to calculate approximate gamma-GT activity. This stain therefore provides a useful method for determining gamma-GT activity when limited cell numbers are available.


Herb-drug interactions: an overview of systematic reviews.
The aim of this overview of systematic reviews (SRs) is to evaluate critically the evidence regarding interactions between herbal medicinal products (HMPs) and synthetic drugs. Four electronic databases were searched to identify relevant SRs. Forty-six SRs of 46 different HMPs met our inclusion criteria. The vast majority of SRs were of poor methodological quality. The majority of these HMPs were not associated with severe herb-drug interactions. Serious herb-drug interactions were noted for Hypericum perforatum and Viscum album. The most severe interactions resulted in transplant rejection, delayed emergence from anaesthesia, cardiovascular collapse, renal and liver toxicity, cardiotoxicity, bradycardia, hypovolaemic shock, inflammatory reactions with organ fibrosis and death. Moderately severe interactions were noted for Ginkgo biloba, Panax ginseng, Piper methysticum, Serenoa repens and Camellia sinensis. The most commonly interacting drugs were antiplatelet agents and anticoagulants. The majority of the HMPs evaluated in SRs were not associated with drug interactions with serious consequences. However, the poor quality and the scarcity of the primary data prevent firm conclusions.


Correlating Salivary Levels of Immunoglobin E and Human Eosinophil Cationic Protein in the Aseer Cohort with Recurrent Apthous Stomatitis.
Repeated Aphthous Stomatitis (RAS) is the most prevalent inflammatory disorder of the oral mucosa, characterized by recurrent emergence of single or numerous painful ulcers. RAS usually affects healthy people without systemic illnesses. There is evidence linking atopy to the progression of this illness. Immunoglobulin E (IgE) and human eosinophil cationic protein (HECP) levels in the saliva of individuals with aphthous stomatitis were assessed as allergy-related indicators. Sixty people were assessed for this study. 30 patients with RAS were included in the patient group, while 30 healthy individuals made up for the control group. Sixty participants' non-stimulated saliva was taken and IgE and HECP were evaluated using enzyme-linked immunosorbent assay (ELISA). Data were analyzed in SPSS 20 through the Mann-Whitney test and p<0.05 was considered significant. The salivary level of HECP was significantly (p 0.05) higher among cases (0.83 0.70) compared to controls (0.170 0.15), whereas the salivary level of IgE was not significantly (p = 0.41) higher among cases (35.60 11.19) compared to controls (67.42 18.34). Even though this study found a positive correlation between elevated HECP levels and RAS, additional research with larger sample sizes is required to identify the biological mechanisms responsible for the observed associations and to include salivary HECP levels in the RAS patient's evaluation.


Cancers in Australia in 2010 attributable to infectious agents.
To estimate the proportion and numbers of cancers in Australia in 2010 attributable to infectious agents. The population attributable fraction (PAF) and number of cancers caused by hepatitis B and C viruses (HBV, HCV), Helicobacter pylori and human immunodeficiency virus (HIV) were calculated using standard formulae incorporating prevalence of infection in the Australian population, the relative risks associated with that infection and cancer incidence. For cancers with very strong associations to the infectious agent (Epstein-Barr virus [EBV], human papillomavirus [HPV] and HIV/Kaposi's sarcoma herpes virus [KSHV]), calculations were based on viral prevalence in the tumour. An estimated 3,421 cancers (2.9% of all cancers) in Australia in 2010 were attributable to infections. Infectious agents causing the largest numbers of cancers were HPV (n=1,706), H. pylori (n=793) and HBV/HCV (n=518). Cancer sites with the greatest number of cancers caused by infections were cervix (n=818), stomach (n=694) and liver (n=483). Cancers with highest proportions attributable to infectious agents were Kaposi's sarcoma (100%), cervix (100%), nasopharynx (87%), anus (84%) and vagina (70%). Infectious agents cause more than 3,000 cancers annually in Australia. Opportunities for cancer prevention through infection control are considerable, even in a 'first world' nation like Australia.


Immunomodulatory effects of recombinant BCG expressing MSP-1C of Plasmodium falciparum on LPS- or LPS+IFN-γ-stimulated J774A.1 cells.
Macrophage phagocytosis is the first line of defense of the innate immune system against malaria parasite infection. This study evaluated the immunomodulatory effects of BCG and recombinant BCG (rBCG) strains expressing the C-terminus of the merozoite surface protein-1 (MSP-1C) of Plasmodium falciparum on mouse macrophage cell line J774A.1 in the presence or absence of lipopolysaccharide (LPS) or LPS + IFN-γ. The rBCG strain significantly enhanced phagocytic activity, production of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, nitric oxide (NO), and inducible nitric oxide synthase (iNOS) as compared with parental BCG strain, and these activities increased in the presence of LPS and LPS+IFN-γ. Furthermore, the rBCG strain also significantly reduced the macrophage viability as well as the rBCG growth suggesting the involvement of macrophage apoptosis. Taken together, these data indicate that the rBCG strain has an immunomodulatory effect on macrophages, thus strengthen the rational use of rBCG to control malaria infection.


Application of oxidation-reduction assay for monitoring treatment of patients with pulmonary tuberculosis.
By oxidation-reduction assay, the viability of Mycobacterium tuberculosis in sputum specimens was evaluated. The technique is based on the Alamar Blue and Malachite Green dyes, which change their color in response to M. tuberculosis growth. The method is simple, permits visual reading of results, and is applicable for laboratories with limited resources.


[Results and complications of CT-guided puncture biopsies with a wide-lumen puncture needle].
The results obtained in 187 CT-guided biopsies using a wide lumen needle are described. Biopsies in the thorax, abdomen, retroperitoneum and in the pelvis have been evaluated. A retrospective analysis of the material has shown an accuracy of 97.8% and sensitivity of 95.8%. The total complication rate was 8%. Amongst 120 biopsies in the thorax, 6.6% developed a pneumothorax. Therapeutic drainage was carried out in fifteen patients.


The role of positive psychological wellbeing in walking speed differences among married and unmarried English older adults.
Objectives: Walking speed has been identified as an important indicator of functional independence and survival among older adults, with marital status being related to walking speed differences. We explored explanatory factors, with a focus on positive psychological wellbeing, in walking speed differences between married and non-married individuals in later life.Methods: We used wave 8 (2016/17) cross-sectional data from adults aged 60-79 years who participated in the English Longitudinal Study of Ageing (n = 3,743). An Oaxaca-Blinder decomposition was used to compute walking speed differences between married and unmarried individuals, and the portion of those differences that could be explained by characteristic differences in those groups, particularly wellbeing.Results: Overall, married individuals had walking speeds that were 0.073 m/s (95% confidence interval: 0.055-0.092 m/s) faster than their unmarried counterparts. This was primarily driven by differences between the married and separated/divorced group, and the widowed group. Included covariates explained roughly 89% of the overall walking speed difference. Positive psychological wellbeing consistently explained a significant portion of walking speed differences, ranging between 7% to 18% across comparisons.Conclusion: Although wealth has been previously found to partially explain walking speed differences by marital status, we found that positive psychological wellbeing also demonstrated pertinence to these differences.


Does prolonged grief treatment foster posttraumatic growth? Secondary results from a treatment study with long-term follow-up and mediation analysis.
Prolonged grief disorder (PGD) is a persistent and disabling kind of grief reaction that can be treated effectively with psychotherapeutic interventions. There has been limited investigation of whether these interventions can also enhance positive outcomes of bereavement, such as posttraumatic growth or benefit finding. As part of secondary analyses in a randomized controlled trial evaluating integrative cognitive-behavioural therapy for PGD (PG-CBT), the posttraumatic growth trajectories in 51 outpatients with clinically relevant prolonged grief symptoms were followed up from baseline up to 1.5 years. Immediate treatment effects on posttraumatic growth in comparison with a waiting list control group were evaluated with univariate ANCOVA. Using mediation analysis, we examined the relation between symptom reduction and the short-term treatment effect on posttraumatic growth. For evaluating long-term outcome stability, the immediately treated group and the delayed treatment group were pooled. PG-CBT significantly fostered growth in patients suffering from PGD, with a controlled medium effect size of Cohen's d = 0.60 (completer analysis). This effect remained stable up to the 1.5-year follow-up. Grief symptom reduction mediated short-term treatment effects on posttraumatic growth. However, growth also partially mediated treatment effects on prolonged grief symptoms. Taken together, PG-CBT was effective in enhancing the participants' perception of posttraumatic growth, but the definite interaction between symptom reduction and posttraumatic growth remains unclear, as both seemed to influence each other's trajectory in the course of treatment. Integrative CBT for prolonged grief disorder also fostered posttraumatic growth. Post-treatment and 1.5-year follow-up effect sizes for posttraumatic growth were moderate. Whether growth-enhancing techniques are useful in grief treatment needs further research.


Physiological changes in laboratory rats caused by different housing.
Males rats of the Wistar strain (Institute of Physiology, CSAV ) were divided into six groups from the 30th to the 200th day of age. Rats (n = 54) were put into cages in various numbers (1, 2, 3, 4, 6 and 8 animals in a cage) and were weighed regularly and the consumption of water and food was measured. At the end of the experiment the animals were killed and the weight of some organs and haematological values were determined. During the experiment the highest weight was attained in animals with 3 or 4 members in one cage. The differences in weight were significant from the 80th day of age. The consumption of diets and water was not influenced significantly by the number of animals in a cage. On the 200th day, the differences between the groups were found in the weight of some organs, and in some haematological values.


Structure-Based Screening of Non-β-Lactam Inhibitors against Class D β-Lactamases: An Approach of Docking and Molecular Dynamics.
The manifestation of class D β-lactamases in the community raises significant concern as they can hydrolyze carbapenem antibiotics. Hence, it is exceptionally alluring to design novel inhibitors. Structure-based virtual screening using docking programs and molecular dynamics simulations was employed to identify two novel non-β-lactam compounds that possess the ability to block different OXA variants. Furthermore, the presence of a nonpolar aliphatic amino acid, valine, near the active site serine, was identified in all OXA variants that can be accounted to block the catalytic activity of OXA enzymes.


A simple open comedone extraction technique for Favre-Racouchot disease.
Favre-Racouchot disease (FRD) is a common disease characterized by solar elastosis and large open comedones and cysts. This disorder mainly affects elderly Caucasian men who have had a great deal of exposure to the sun. Patients having FRD continuously develop numerous large open comedones that could cause a cosmetic problem. We performed a simple and effective extraction technique for these open comedones using a standard dissecting forceps. This technique is well tolerated and no complications were observed. We suggest this comedone extraction technique as a simple alternative treatment for this common cosmetic problem.


Renal failure following unilateral nephrectomy in Wilms tumour.
A three year old male child with left sided Wilms tumour with vena caval involvement developed acute renal failure following nephrectomy. The likely etiology of acute renal shut down postoperatively and the relevance of differences in venous anatomy of the right and left kidneys in such a situation are discussed.


Treatment of paraplegic sheep with hyperbaric oxygen.
The results of the use of hyperbaric oxygen therapy to control the onset of paraplegia after recent spinal cord injury in sheep are described. This preliminary report suggest that hyperbaric oxygen therapy instituted within two hours of the injury will result in improved motor recovery.


Management of severe rhinophyma with sculpting surgical decortication.
Rhinophyma, a progressive, disfiguring disease of the nose, is considered the final stage of acne rosacea. In its early stages, rhinophyma can be managed with medical treatment using isotretinoin or oral antibiotics (metronidazole). However, severe cases usually are refractory to medical approaches. Surgical therapies to treat these severe refractory cases have been described. This report describes a simple, safe, efficient, and cost-effective approach to the treatment of severe rhinophyma using a scalpel and the electroscalpel, instruments readily available in every operating room. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


TP53 mutations predict for poor outcomes in patients with newly diagnosed aggressive B-cell lymphomas in the current era.
Genetic subgroups of diffuse large B-cell lymphoma (DLBCL) have been identified through comprehensive genomic analysis; however, it is unclear whether this can be applied in clinical practice. We assessed whether mutations detected by clinical laboratory mutation analysis (CLMA) were predictive of outcomes in patients with newly diagnosed DLBCL/high-grade B-cell lymphoma (HGBL). Patients diagnosed from 2018 to 2022 whose biopsy samples were subjected to CLMA and who received rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone or rituximab plus etoposide, prednisolone, vincristine, cyclophosphamide, and doxorubicin were analyzed for overall/complete response rate (ORR/CRR) and estimated progression-free/overall survival (PFS/OS). CLMA was successfully performed in 117 of 122 patient samples (96%), with a median turnaround time of 17 days. Median duration of follow-up was 31.3 months. Of the mutations detected in ≥10% of the samples, only TP53 was associated with both progression and death at 2 years. TP53 mutations were detected in 36% of tumors, and patients with TP53 mutations experienced significantly lower ORR (71% vs 90%; P = .009), CRR (55% vs 77%; P = .01), 2-year PFS (57% vs 77%; P = .006), 2-year OS (70% vs 91%; P = .001), and median OS after relapse (6.1 months vs not yet reached; P = .001) as than those without TP53 mutations. Furthermore, patients with TP53 loss-of-function (LOF) mutations experienced lower rates of 2-year PFS/OS than those with non-LOF mutations and inferior or near-inferior 2-year PFS if harboring high-risk clinicopathologic features. TP53 mutations identified through CLMA can predict for inferior outcomes in patients with newly diagnosed DLBCL/HGBL. Results of CLMA can be used in real time to inform prognosis and/or identify candidates for clinical trials.


Antifungal and antiviral chemotherapy.
Fungal and viral infections are difficult to treat, since fungal infections commonly rebound after suppression by the antifungal agent and current antiviral drugs are only virustatic, allowing the virus to reassert its pathogenicity if not eliminated by the host defenses. In addition, fungal infections commonly are associated with significant biofilms, retarding drug penetration, and the fluid nature of the oral cavity does not promote drug-fungus contact for long periods of time. Both mycotic and viral pathogens are developing sophisticated methods to elude the toxic effects of drugs intended to eliminate their existence. The drug therapy of oral fungal and viral infections is therefore limited but occasionally successful (more with fungal than viral infections) and is often relegated to palliative care. The specter of drug resistance and its promotion by prolonged, repetitive and frivolous use must always be foremost in the clinician's mind.


Surfactant supplementation in the preterm rabbit: effects of applied volume on compliance and survival.
Preterm rabbit neonates, delivered on the 27th day of gestation, were treated before the first breath with a tracheal instillation of ESA, a suspension of lipids extracted from pulmonary surfactant. Lung compliance development and neonatal survival were studied after treatment to determine the effect of suspension volume and concentration. Compliance development was not dependent on either delivery volume or suspension concentration, but rather on the quantity of phospholipids instilled. Indices of improved expansion and stability demonstrated a dose response relationship up to 1.1 mg phospholipid per g lung. Larger doses neither improved nor detracted from the response. The ability of the neonates to survive and adapt to an air breathing environment was influenced by the instillation volume. Viability was reduced when the treatment volume exceeded 16% of the functional residual capacity. Prophylactic surfactant supplementation therapy may be useful in preventing the development of neonatal respiratory distress syndrome. Surfactant extract may prove acceptable for both prophylaxis and treatment of established respiratory distress syndrome.


Hepatitis C virus infection among hemodialysis patients in the eastern region of saudi arabia.
The prevalence of anti-HCV antibodies among hemodialysis patients in the Eastern region of Saudi Arabia was 42.8%. This is comparable to the observations from the Central region of the country. Repeated blood transfusions of non-screened HCV blood seems to be the major factor responsible for this high prevalence. Clinical liver disease was infrequent in the early phase of the infection.


Brain stem auditory, visual and somatosensory evoked potentials in leukodystrophies.
Brain stem auditory (BAERs), visual (VEPs) and somatosensory evoked responses (SEPs) were recorded in 12 patients with Pelizaeus-Merzbacher leukodystrophy (PMD), three with adrenoleukodystrophy (ALD) and three with metachromatic leukodystrophy (MLD). All the 3 evoked responses were abnormal in all patients except normal VEPs and SEPs in a patient with early ALD. In most patients wave I with and without wave II were the only components of the BAERs that remained, subsequent components (waves III-VII) were absent. VEPs were severely altered; either no identifiable response to flash or pattern reversal stimuli could be recorded or the major components were significantly delayed in latency. Short latency SEPs following median nerve stimulation usually showed a normally recorded Erb's potential (N10), but an absence or marked attenuation of cervical (N14) and early scalp components (N19 and P22) or the occurrence of the scalp components with a significant delay. Multimodality evoked responses provide more information regarding the functional integrity of several afferent systems in patients with white matter disorders.


UHRF1 regulation of the Keap1-Nrf2 pathway in pancreatic cancer contributes to oncogenesis.
The cellular defence protein Nrf2 is a mediator of oncogenesis in pancreatic ductal adenocarcinoma (PDAC) and other cancers. However, the control of Nrf2 expression and activity in cancer is not fully understood. We previously reported the absence of Keap1, a pivotal regulator of Nrf2, in ∼70% of PDAC cases. Here we describe a novel mechanism whereby the epigenetic regulator UHRF1 suppresses Keap1 protein levels. UHRF1 expression was observed in 20% (5 of 25) of benign pancreatic ducts compared to 86% (114 of 132) of pancreatic tumours, and an inverse relationship between UHRF1 and Keap1 levels in PDAC tumours (n = 124) was apparent (p = 0.002). We also provide evidence that UHRF1-mediated regulation of the Nrf2 pathway contributes to the aggressive behaviour of PDAC. Depletion of UHRF1 from PDAC cells decreased growth and enhanced apoptosis and cell cycle arrest. UHRF1 depletion also led to reduced levels of Nrf2-regulated downstream proteins and was accompanied by heightened oxidative stress, in the form of lower glutathione levels and increased reactive oxygen species. Concomitant depletion of Keap1 and UHRF1 restored Nrf2 levels and reversed cell cycle arrest and the increase in reactive oxygen species. Mechanistically, depletion of UHRF1 reduced global and tumour suppressor promoter methylation in pancreatic cancer cell lines, and KEAP1 gene promoter methylation was reduced in one of three cell lines examined. Thus, methylation of the KEAP1 gene promoter may contribute to the suppression of Keap1 protein levels by UHRF1, although our data suggest that additional mechanisms need to be explored. Finally, we demonstrate that K-Ras drives UHRF1 expression, establishing a novel link between this oncogene and Nrf2-mediated cellular protection. Since UHRF1 over-expression occurs in other cancers, its ability to regulate the Keap1-Nrf2 pathway may be critically important to the malignant behaviour of these cancers.


Electrocardiographic changes in normal and abnormal canine pregnancy.
The aim of this study was to describe the canine electrocardiographic changes in the course of normal and abnormal pregnancy. Twenty-three Brucellosis-negative pregnant bitches were retrospectively classified as normal (n = 12) or abnormal (n = 11). A control group of non-pregnant dioestrous bitches (n = 10) was also included. Normal pregnant females delivered healthy puppies at term while abnormal animals interrupted their pregnancy between days 52-60 (from estimated luteinizing hormone peak) or presented perinatal litter death higher than 60%. All the bitches were electrocardiographically evaluated every 10 days from day 0 to day 65 of the oestrous cycle, to parturition or abortion. Percentage heart rate change increased 31.3% from day 40 to 60 in normal gestation while it decreased -1.8% in dioestrous bitches, although it did not change in the abnormal group (p < 0.01). In the abnormal pregnant group but not in the others, percentage QRSa change fell to -34% on day 60 (p < 0.01). At the same time point, percentage QRSd change was 6.2% vs -4.9% in normal gestations and dioestrous animals, respectively (p < 0.05). Corrected QT interval augmented from day 40 onwards up to 9.9% and 4.3% in the normal pregnant and dioestrous groups, respectively, while it remained unchanged in abnormal gestations (p < 0.05). It is concluded that during normal canine pregnancy, some electrocardiographic parameters begin changing from day 40 onwards, and that pathological gestations differ from normality from day 30. The use of electrocardiography in canine obstetrics might contribute to identify abnormal outcomes before they become clinically evident.


Euvolemic hyponatremia in cancer patients. Report of the Hyponatremia Registry: an observational multicenter international study.
Hyponatremia secondary to SIADH is frequent in cancer patients and potentially deleterious. The aim of this sub-analysis of the Hyponatremia Registry database is to analyze current diagnostic and therapeutic management practices in cancer patients with SIADH. We analyzed 358 cancer patients who had serum sodium concentration ([Na+]) ≤ 130 mEq/L and a clinical diagnosis of SIADH from 225 sites in the USA and EU. Precise diagnostic testing was performed in only 46%. Almost 12% of all patients did not receive any hyponatremia treatment. The most frequent therapies were fluid restriction (20%), isotonic saline (14%), fluid restriction/isotonic saline (7%), tolvaptan (8%), and salt tablets (7%). Hypertonic saline was used in less than 3%. Tolvaptan produced the greatest median rate of [Na+] change (IQR) (3.0 (4.7) mEq/L/day), followed by hypertonic saline (2.0(7.0) mEq/L/day), and fluid restriction/isotonic saline (1.9(3.2) mEq/L/day). Both fluid restriction and isotonic saline monotherapies were significantly less effective (0.8(2.0) mEq/L/day and 1.3(3.0) mEq/L/day, respectively) and were associated with clinically relevant rates of treatment failure. Only 46% of patients were discharged with [Na+] ≥ 130 mEq/L. Overly rapid correction of hyponatremia occurred in 11.7%. Although essential for successful hyponatremia management, appropriate diagnostic testing is not routinely performed in current practice. The most frequently employed monotherapies were often ineffective and sometimes even aggravated hyponatremia. Tolvaptan was used less often but showed significantly greater effectiveness. Despite clear evidence that hyponatremia is associated with poor outcome in oncology patients, most patients were discharged still hyponatremic. Further studies are needed to assess the beneficial impact of hyponatremia correction with effective therapies.


Mid-infrared reflectivity of experimental atheromas.
We report that the pathologic components present within the atheromatous plaques of ApoE knock-out mice can reflect significant amounts of mid-infrared (mid-IR) light. Furthermore, the reflected light spectra contained the unique signatures of a variety of biologic features including those found in unstable or "vulnerable" plaque. This discovery may represent a unique opportunity to develop a new intravascular diagnostic modality that can detect and characterize sites of atherosclerosis.


Recombinant expression of Munc18c in a baculovirus system and interaction with syntaxin4.
Two protein families that are critical for vesicle transport are the Syntaxin and Munc18/Sec1 families of proteins. These two molecules form a high affinity complex and play an essential role in vesicle docking and fusion. Munc18c was expressed as an N-terminally His-tagged fusion protein from recombinant baculovirus in Sf9 insect cells. His-tagged Munc18c was purified to homogeneity using both cobalt-chelating affinity chromatography and gel filtration chromatography. With this simple two-step protocol, 3.5 mg of purified Munc18c was obtained from a 1L culture. Further, the N-terminal His-tag could be removed by thrombin cleavage while the tagged protein was bound to metal affinity resin. Recombinant Munc18c produced in this way is functional, in that it forms a stable complex with the SNARE interacting partner, syntaxin4. Thus we have developed a method for producing and purifying large amounts of functional Munc18c--both tagged and detagged--from a baculovirus expression system. We have also developed a method to purify the Munc18c:syntaxin4 complex. These methods will be employed for future functional and structural studies.


The Role of Allergy Testing in Eosinophilic Esophagitis.
Eosinophilic esophagitis (EoE) is defined as a chronic, immune/antigen-mediated esophageal disease that can lead to symptoms of esophageal dysfunction. This disease is seen in both children and adults. Approximately 70% of patients with EoE have food antigen sensitization or other atopic conditions, suggesting an allergic etiology in the pathogenesis of the disease. The role of allergy testing to identify foods that lead to EoE is unclear. Three types of allergy tests currently exist: skin prick testing, atopy patch testing, and immunoassays for serum food-specific immunoglobulin E. It is important for gastroenterologists to work in conjunction with allergist colleagues in the care of patients with EoE, particularly in the management of comorbid atopic conditions.