[Friedreich's spinal ataxia following measles? Overview of central nervous system diseases caused by measles - (preliminary report)].
A case of Friedreich's ataxia of the spine with intranuclear bodies in the cerebrum and stem was found in a 43-years old man. The disease is possibly the result of measles contracted when the patient was seven. This is the first observation of this type known to us; Friedreich's spinal ataxia has until now been believed to exist only hereditary ("hereditary spinal ataxia").


Expression of the bovine heart mitochondrial ADP/ATP carrier in yeast mitochondria: significantly enhanced expression by replacement of the N-terminal region of the bovine carrier by the corresponding regions of the yeast carriers.
To characterize the transport mechanism mediated by the mammalian mitochondrial ADP/ATP carrier (AAC), we tried to express bovine heart mitochondrial AAC (bhAAC) in Saccharomyces cerevisiae. The open reading frame of the bhAAC was introduced into the haploid strain WB-12, in which intrinsic AAC genes were disrupted. Growth of the transformant was very low in glycerol medium, and a little amount of bhAAC was detected in the mitochondrial membrane. For improvement of bhAAC expression in WB-12, we introduced DNA fragments encoding chimeric bhAACs, in which the N-terminal region of the bhAAC extending into the cytosol was replaced by the corresponding regions of the type 1 and type 2 yeast AAC isoforms (yAAC1 and yAAC2). These transformants grew well, and the amounts of the chimeric bhAACs in their mitochondria were as high as that of yAAC2. The carriers expressed showed essentially the same ADP transport activities as that of AAC in bovine heart mitochondria.


Impact of obstructive sleep apnoea on left ventricular mass and global function.
Obstructive sleep apnoea syndrome (OSAS) might be a cause of heart failure. The present study aimed to assess left ventricular mass and myocardial performance index (MPI) in OSAS patients. A total of 67 subjects without any cardiac or pulmonary disease, referred for evaluation of OSAS, had overnight polysomnography and echocardiography. According to apnoea-hypopnoea index (AHI), subjects were classified into three groups: mild OSAS (AHI: 5-14; n = 16), moderate OSAS (AHI: 15-29; n = 18), and severe OSAS (AHI: > or = 30; n = 33). Thickness of interventricular septum (IVS) and posterior wall (LVPW) were measured by M-mode, along with left ventricular mass (LVM) and LVM index (LVMI). Left ventricular MPI was calculated as (isovolumic contraction time+isovolumic relaxation time)/aortic ejection time by Döppler echocardiography. There were no differences in age or body mass index among the groups, but blood pressures were higher in severe OSAS compared with moderate and mild OSAS. In severe OSAS, thickness of IVS (11.2+/-1.1 mm), LVPW (11.4+/-0.9 mm), LVM (298.8+/-83.1 g) and LVMI (144.7+/-39.8 g x m(-2)) were higher than in moderate OSAS (10.9+/-1.3 mm; 10.8+/-0.9 mm; 287.3+/-74.6 g; 126.5+/-41.2 g x m(-2), respectively) and mild OSAS (9.9+/-0.9 mm; 9.8+/-0.8 mm; 225.6+/-84.3 g; 100.5+/-42.3 g x m(-2), respectively). In severe OSAS, MPI (0.64+/-0.14) was significantly higher than in mild OSAS (0.50+/-0.09), but not significantly higher than moderate OSAS (0.60+/-0.10). In conclusion, severe and moderate obstructive sleep apnoea syndrome patients had higher left ventricular mass and left ventricular mass index, and also left ventricular global dysfunction.


Micro and macroarchitectural changes at the tibia after botulinum toxin injection in the growing rat.
The aim of this study was to analyze bone microarchitecture and macroarchitecture of tibia in a disuse model in growing rats. Eight-weeks-old Copenhagen rats were injected intramuscularly with 1.5 units BTX in the quadriceps muscle of the right hind limb. Saline injection was done at the left hind limb to serve as control. Five rats were killed at day 1 and represented the baseline group (D1), 5 rats were killed at day 14 (D14), 5 at day 21 (D21), 5 at day 28 (D28) and 5 at day 35 (35). For each group, muscle surface, parameters of bone microarchitecture and macroarchitecture (including length, width and curvature of the tibia) were measured using microtomography. Paralysis occurred as soon as day 2. At the left hind limb, muscle surface area, cortical thickness, cross sectional total area and growth in length significantly increased during the time study. At the right hind limb, muscle surface area, bone trabecular volume, and cortical thickness decreased as soon as day 14 associated with an increased cortical porosity. Growth in length did not differ from left side; cross sectional total area did not increase and the diaphyseal cross section acquired a more rounded shape. There was no modification of the curvature between right and left hind limbs during the time study. In this murine model of unilateral muscle paralysis in growing animals, we showed a rapid muscle loss leading to a decreased growth in width; however growth in length and curvature were unaltered.


Adaptive servo-ventilation therapy improves cardiac sympathetic nerve activity in patients with heart failure.
This study investigated whether abnormal cardiac sympathetic nerve activity (SNA) is associated with the severity of central sleep apnoea (CSA) and whether adaptive servo-ventilation (ASV) therapy can improve cardiac SNA in heart failure (HF) patients with predominant CSA. Overnight polysomnography was conducted to diagnose CSA. Cardiac SNA was analysed by [123I]metaiodobenzylguanidine scintigraphy in 26 consecutive HF patients with predominant CSA. Of the 26 patients, 10 agreed to ASV therapy. Cardiac SNA was analysed 6 months after initiating ASV based on a non-randomized protocol. The apnoea-hypopnoea index and central apnoea index were significantly correlated with the washout rate (WR) and a delayed heart to mediastinal (H/M) ratio, suggesting that SNA is associated with abnormal breathing patterns. The WR, H/M ratio, plasma BNP level, and LVEF were significantly improved (WR, 40.0 ± 11.6% vs. 34.6 ± 11.4%, P = 0.046; H/M ratio, 1.5 ± 0.1 vs.1.8 ± 0.3, P = 0.013; ln BNP, 5.4 ± 1.0 vs. 4.6 ± 1.2, P = 0.007; and LVEF, 43.8 ± 10.4% vs. 47.0 ± 10.6%, P < 0.001) in the ASV group patients, but not in the non-ASV group patients. Multiple linear regression analyses showed that a decreased WR was strongly associated with an increased LVEF (coefficient = -0.454, P = 0.013). Abnormal cardiac SNA could be significantly correlated with the severity of CSA in HF patients. ASV therapy might improve cardiac function in these patients by partially mediating cardiac SNA regulation.


Effects of drugs on pigeon erythrocyte membrane and asymmetric control or adenylate cyclase by the lipid bilayer.
In pigeon erythrocyte membrane, the beta-adrenergic receptor and the enzyme adenylate cyclase can be uncoupled in two different ways depending on the type of drug used. Cationic drugs: chlorpromazine, methochlorpromazine, tetracaine, n-octylamine and a neutral alcohol, octanol, abolished alprenolol receptor binding ability and in the same range of concentration of the drug, sensitized adenylate cyclase to fluoride or Gpp(NH)p stimulation. Anionic drugs: di- and trinitro-phenols, indomethacin and octanoic acid did not affect the total number of beta-adrenergic receptor sites and, with the exception of trinitrophenol, did not change the association constant for alprenolol but they abolished the stimulation of adenylate cyclase by isoproterenol, fluoride or Gpp(NH)p. These modifications of the adenylate cyclase system occurred in a range of drug concentration where cell shape and protection against hemolysis were also affected. As chemical composition varies widely from one drug to another, it is suggested that these effects are largely nonspecific and mediated by the lipid bilayer. They are probably related to a preferential sidedness of action of the drugs in the lipid bilayer, displaying the role of an asymmetric control of the adenylate cyclase system in the membrane by the two halves of this bilayer.


Reducing Cascading Failure Risk by Increasing Infrastructure Network Interdependence.
Increased interconnection between critical infrastructure networks, such as electric power and communications systems, has important implications for infrastructure reliability and security. Others have shown that increased coupling between networks that are vulnerable to internetwork cascading failures can increase vulnerability. However, the mechanisms of cascading in these models differ from those in real systems and such models disregard new functions enabled by coupling, such as intelligent control during a cascade. This paper compares the robustness of simple topological network models to models that more accurately reflect the dynamics of cascading in a particular case of coupled infrastructures. First, we compare a topological contagion model to a power grid model. Second, we compare a percolation model of internetwork cascading to three models of interdependent power-communication systems. In both comparisons, the more detailed models suggest substantially different conclusions, relative to the simpler topological models. In all but the most extreme case, our model of a "smart" power network coupled to a communication system suggests that increased power-communication coupling decreases vulnerability, in contrast to the percolation model. Together, these results suggest that robustness can be enhanced by interconnecting networks with complementary capabilities if modes of internetwork failure propagation are constrained.


The contribution of laparoscopy in evaluation of penetrating abdominal wounds.
Penetrating abdominal wounds are traditionally explored by laparotomy. We investigated prospectively the role of laparoscopy within a defined protocol for management of penetrating abdominal wounds to determine its safety and advantages over traditional operative management. The study inclusion criteria were: stab and gun shot abdominal wounds, including junction zone injuries; stable vital signs; and absence of contraindications for laparoscopy. Diagnostic end points included detection of peritoneum or diaphragm violation, visceral injuries, and other indications for laparotomy. Systematic examination was undertaken using a multiport technique whenever the peritoneum or diaphragm had been violated. All repairs were done by open operation. A total of 40.6% of patients with penetrating trauma fulfilled study criteria (52 patients). Of these, 33% had no peritoneal penetration; 29% had no visceral injuries despite violation of peritoneum or diaphragm; 38% had visceral injuries, of which 40% (mainly liver and omentum) required no intervention. Twelve patients (23% of total) had open repairs. No missed injuries or death occurred in the study. Overall, 77% of penetrating injuries with stable vital signs avoided exploratory laparotomy. Compared with National Trauma Data Bank information for patients with the same Injury Severity Scores, hospitalization was reduced by more than 55% for the entire series. Laparoscopy for penetrating abdominal injuries in a defined set of conditions was safe and accurate, effectively eliminating nontherapeutic laparotomy and shortening hospitalization.


Potential efficacy of local therapy for progressive lesions after nivolumab in patients with recurrent or metastatic squamous cell carcinoma of the head and neck.
In recurrent or metastatic squamous cell carcinoma of the head and neck (R/M SCCHN), local therapy (LT) such as surgery or radiotherapy can be treatment options for improved survival or quality of life. To date, however, few reports have addressed the efficacy of LT for sites of disease progression after immune checkpoint inhibitors, including other cancers. We conducted a retrospective analysis of patients with R/M SCCHN originating from the oral cavity, oropharynx, hypopharynx, and larynx and treated with nivolumab. We extracted patients undergoing salvage LT or palliative radiotherapy (RT) to the selected progressive lesion at any time after initiation of nivolumab. Twenty-four patients received LT. Salvage LT was performed in 9 (37.5%) patients, including surgery and definitive RT in 5 and 4 patients, respectively. Palliative RT was performed in 15 (62.5%) patients. LT was provided in 10 (41.7%) patients for oligoprogressive disease. Twelve (50.0%) patients received subsequent systemic therapy immediately after LT. Classification based on patient treatment divided the population into four subgroups with different prognoses (salvage LT followed by subsequent systemic therapy [n = 3], salvage LT alone [n = 6], palliative RT followed by subsequent systemic therapy [n = 9], and palliative RT alone [n = 6]). Median OS in this order was 24.5, 9.0, 7.3, and 2.4 months (p = 0.001). All patients in the salvage LT followed by subsequent systemic therapy group continued nivolumab. In R/M SCCHN patients who have received nivolumab, salvage LT for the selected progressive lesion with continuation of nivolumab potentially provides an excellent survival prognosis.


A new generation of progesterone receptor modulators.
Progesterone receptor (PR) modulators have evolved both structurally and mechanistically over the past half-century. Classical steroidal PR agonists continue to play an important role in women's health such as in oral contraception and post-menopausal hormone therapy whereas steroid-based PR antagonists and selective PR modulators are being evaluated clinically in a wide range of gynecologic conditions. This review will focus primarily on the newer generation of PR modulators derived from structurally unique chemical scaffolds. For example, tanaproget (TNPR) is described as a non-steroidal PR agonist with high affinity and selectivity for the PR that is significantly more potent than many of its steroidal counterparts in a variety of pre-clinical efficacy models. Similarly, we present numerous examples of unique non-steroidal PR antagonists in various stages of characterization and development. A basic understanding of the structural determinants for high affinity binding of these new PR modulators to the PR ligand-binding domain (LBD) is also discussed. Finally, as the biology of the PR becomes further defined, we speculate on the future development of novel PR modulators.


Recent advances in systemic lupus erythematosus and microbiota: from bench to bedside.
Systemic lupus erythematosus (SLE) is a complicated autoimmune disease affecting multiple systems and organs. It is highly heterogeneous, and it preferentially affects women at childbearing age, causing worldwide social burden. The pathogenesis of SLE mostly involves genetic predisposition, epigenetic dysregulation, overactivation of the immune system, and environment factors. Human microbiome, which is mostly composed of microbiota colonized in the gut, skin, and oral cavity, provides a natural microbiome barrier against environmental risks. The past decade of research has demonstrated a strong association between microbiota and metabolic diseases or gastrointestinal diseases. However, the role of microbiota in autoimmunity remains largely unknown until recently, when the technological and methodological progress facilitates further microbiota research in SLE. In this review, the latest research about the role and mechanisms of microbiota in SLE and the advances in the development of diagnostic and therapeutic strategies based on microbiota for SLE were summarized.


New methods for implanting plastic tubes for 192iridium brachytherapy using steel needles.
A revised method for implanting plastic tubes used to carry 192iridium for brachytherapy is described. Solid malleable stainless steel needles are machined at one end with a tapered screw thread, on to which the plastic tubing can be placed for pulling through tissue. The technique has proved to be both simpler and quicker than the standard 'Amersham' method. Since the solid needles may be moulded, it is easier to implant relatively inaccessible areas such as axillary, intrathoracic and paraspinal sites. For sites with no clear exit point for the needles, a large diameter steel needle is described, which enables the plastic tubes to be inserted with less trauma to the tissues.


Detection of fetomaternal hemorrhage.
The prevention of Rhesus D alloimmunization through Rh immune globulin (RhIg) administration is the major indication for the accurate detection and quantification of fetomaternal hemorrhage (FMH). In the setting of D incompatibility, D-positive fetal cells can sensitize the D-negative mother, resulting in maternal anti-D alloantibody production. These anti-D alloantibodies may lead to undesirable sequelae such as hemolytic disease of the newborn (HDN). Since the widespread adoption of FMH screening and RhIg immunoprophylaxis, the overall risk of Rh alloimmunization and infant mortality from HDN has substantially decreased. The rosette screen, the initial test of choice, is highly sensitive in qualitatively detecting 10 mL of fetal whole blood in the maternal circulation. As the screen is reliant on the presence of the D antigen to distinguish fetal from maternal cells, it cannot be used to detect FMH in D-positive mothers or in D-negative mothers carrying a D-negative fetus. The Kleihauer-Betke acid-elution test, the most widely used confirmatory test for quantifying FMH, relies on the principle that fetal RBCs contain mostly fetal hemoglobin (HbF), which is resistant to acid-elution whereas adult hemoglobin is acid-sensitive. Although the Kleihauer-Betke test is inexpensive and requires no special equipment, it lacks standardization and precision, and may not be accurate in conditions with elevated F-cells. Anti-HbF flow cytometry is a promising alternative, although its use is limited by equipment and staffing costs. Hematology analyzers with flow cytometry capabilities may be adapted for fetal cell detection, thus giving clinical laboratories a potentially attractive automated alternative for quantifying FMH.


A radioactive assay for the physiological activity of the tryptophan synthetase alpha subunit in crude extracts of Escherichia coli.
A modified assay has been devised for the physiological reaction, indole-3-glycerol phosphate to Trp, of the enzyme tryptophan synthetase. The assay may be applied to crude bacterial extracts, and is based on the measurement of incorporation of radioactivity from [3H]Ser into Trp. Comparison with previous colorimetric assays indicates an improvement in sensitivity of about 30-fold, and advantages in terms of sample economy and simplified manipulation.


A bis-pyridinium fullerene derivative induces apoptosis through the generation of ROS in BCR-ABL-positive leukemia cells.
A fusion protein, Breakpoint cluster region-Abelson (BCR-ABL) is responsible for the development of chronic myeloid leukemia (CML) and acute lymphocytic leukemia (ALL). Inhibitors against BCR-ABL are effective for the treatment of leukemia; however, a gatekeeper mutation (T315I) in BCR-ABL results in resistance to these inhibitors, which markedly impedes their efficacy. We herein demonstrated that a bis-pyridinium fullerene derivative (BPF) significantly induced apoptosis in human CML-derived K562 cells and ALL-derived SUP-B15 cells via the generation of reactive oxygen species (ROS). BPF reduced the expression of Bcr-Abl mRNA by inhibiting expression of c-Myc through ROS production. BPF also accelerated protein degradation of BCR-ABL through ROS production. Furthermore, BPF down-regulated the expression of not only BCR-ABL but also T315I-mutated BCR-ABL in ROS-dependent manner. As a result, BPF effectively induced apoptosis in transformed Ba/F3 cells expressing both BCR-ABL and T315I-mutated BCR-ABL. Collectively, these results indicate the potential of BPF as an effective leukemia drug that overcomes resistance to BCR-ABL inhibitors.


Facile synthesis of Fe3O4 nanoparticles/reduced graphene oxide sandwich composites for highly efficient microwave absorption.
Component regulation and microstructure design are two effective strategies to adjust electromagnetic parameters and improve the microwave absorption performance of materials. In this study, a facile synthesis strategy consisting of ultrasonic dispersion, blast drying, and roasting is proposed to build a sandwich-like graphene-based absorbent, in which Fe3O4 nanoparticles with adjustable content are sandwiched uniformly between reduced graphene oxide nanosheets. The sandwich structure can form multiple interfaces, prevent the aggregation of nanoparticles, facilitate interface polarization, and endow the material with multiple electromagnetic loss mechanisms, which is very beneficial for impedance matching and microwave attenuation. Notably, the effective absorption bandwidth achieves 5.7 GHz, and the minimum reflection loss value is -49.9 dB. In addition, the synthesis process is simple and suitable for large-scale production and possible industrial applications. Thus, this facile route to fabricate sandwich-like graphene-based absorbents provides new ideas and approaches for designing new graphene-based nanocomposites.


Human immunodeficiency virus infection and its association with sarcopenia.
Presarcopenia and sarcopenia were evaluated in HIV-infected individuals and in healthy elderly controls according to the consensus definitions of the European Working Group on Sarcopenia in Older People. Bioelectrical impedance, a hydraulic hand dynamometer, and gait speed were used to evaluate muscle mass, muscle strength, and physical performance, respectively. Adjusted and unadjusted binary logistic regression predicted the risk of sarcopenia. Predictor contribution was assessed by the Wald test. Significance was established at p≤0.05. The HIV-infected group consisted of 33 patients on treatment (42.4% women; mean age 59±7 years; mean BMI 25±6kg/m(2); viral load undetectable in 30 cases). The HIV-uninfected group consisted of 60 individuals (71.7% women; mean age 70±7 years; mean BMI 28±6kg/m(2)). Of the controls, 4 (6.7%) individuals had presarcopenia and 4 (6.7%) sarcopenia compared to 4 (12.1%) and 8 (24.2%), respectively, in the HIV-infected group. The HIV-infected patients had a 4.95 higher risk (95% CI: 1.34-18.23) for sarcopenia compared to the controls. It should be pointed out that the control group was on average 10 years older. This risk increased further (RR=5.20; 95% CI: 1.40-19.20) after adjusting for age and BMI. HIV-infected patients were shown to be at a greater risk of sarcopenia, an indicator of frailty, even following adjustment for age and BMI.


Effectiveness of tocilizumab in non-intubated cases with COVID-19: a systematic review and meta-analysis.
Tocilizumab, a monoclonal IL-6 receptor blocker, is an effective agent for severe-to-critical cases of COVID-19; however, its target patients for the optimum use need to be detailed. We performed a systematic review and meta-analysis to define its effect among severely ill but non-intubated cases with COVID-19. We searched PubMed, Scopus, Web of Science, MEDLINE, Cochrane Central Register of Controlled Trials (CENTRAL), Medrxiv, and Biorxiv until February 13, 2022, for non-intubated cases, and included randomized-controlled trials (RCT) based on bias assessment. The primary outcomes were the requirement of invasive mechanical ventilation and mortality. Random effect and fixed-effect models were used. The heterogeneity was measured using the χ2 and I2 statistics, with χ2 p ≤ 0.05 and I2 ≥ 50% indicating the presence of significant heterogeneity. We registered the study to the International Prospective Register of Systematic Reviews (PROSPERO) with the registration number CRD42021232575. Among 261 articles, 11 RCTs were included. The pooled analysis of the 11 RCTs demonstrated that the rate of mortality was significantly lower in the tocilizumab group than in the control group (20.0% and 24.2%, OR: 0.84, 95% CI 0.73-0.96, and heterogeneity I2 = 0%. p = 0.82.). The mechanical ventilation rate was lower in the tocilizumab group than the control group (27% vs 35.2%, OR: 0.76, 95% CI 0.67-0.86, and heterogeneity I2 = 6%. p = 0.39). Among non-intubated severe COVID-19 cases, tocilizumab reduces the risk of invasive mechanical ventilation and mortality compared to standard-of-care treatment.


A multicenter, double-blind, randomized trial on the bleeding profile of a drospirenone-only pill 4 mg over nine cycles in comparison with desogestrel 0.075 mg.
A typical compliance problem in the use of traditional progestin-only pills is the irregular bleeding pattern and the strict daily intake. Desogestrel 75 mg has a 12-h missed-pill window; however, its poor cycle control limits a more common use. A drospirenone (DRSP)-only pill was developed to improve the bleeding profile. A phase III study in healthy women aged 18-45 years was performed to compare the bleeding profile and safety of a DRSP-only pill in a regime of 24 days of 4 mg of DRSP tablets followed by 4 days of placebo versus desogestrel 0.075 mg per day continuously over nine cycles. A total of 858 women with 6691 drospirenone and 332 women with 2487 desogestrel treatment cycles were analyzed. The primary end point was the proportion of women with unscheduled bleeding/spotting in each cycle from cycles 2 to 9 and cumulative in cycles 2-4 and cycles 7-9. In each cycle, up to cycle 7, the proportion of women with unscheduled bleeding was statistically significantly lower in the DRSP group than in the DSG group (p = 0.0001, Chi-square test). This report describes the improvement in bleeding profile of women using the new DRSP-only oral contraceptive in comparison to DSG, providing a better quality of live and adherence to the contraceptive method. EudraCT Registration Number: 2011-002396-42.


Effects of low-level microwave irradiation on hippocampal and frontal cortical choline uptake are classically conditionable.
In previous research, we found that sodium-dependent high-affinity choline uptake in the hippocampus and frontal cortex of the rat was lowered after acute (45 min) exposure to low-level 2450-MHz pulsed microwaves (power density 1 mW/cm2; average whole body specific absorption rate, 0.6 W/kg; 2 mu sec pulses, 500 pps). In the present experiment, we investigated developments of tolerance and classical conditioning to these effects of microwaves. Rats were exposed to microwaves in cylindrical waveguides in 10 daily sessions (45 min per session). In an 11th session, we subjected the rats to either microwave (study of tolerance) or sham exposure (study of conditioned effect) for 45 min, and immediately measured choline uptake in the hippocampus and frontal cortex. We found that tolerance, a decrease in response to microwaves, developed to the effect of microwaves on choline uptake in the hippocampus, but not in the frontal cortex. Conditioned effects were also observed: an increase in choline uptake in the hippocampus and a decrease in uptake in the frontal cortex. These data suggest that the effects of microwaves on choline uptake in the hippocampus and frontal cortex are classically conditionable, probably to cues in the exposure environment.


[Allergic diagnosis of abortive chlamydial infection in the goat (author's transl)].
A delayed hypersensitivity test was used for chlamydiosis diagnosis in four goat flocks. The skin-test was performed in the neck by inoculating 100 microgram of purified Chlamydia from yolk sac or McCoy cells. The reactions were read 72 h after inoculation. The two antigens gave the same results. The number of doubtful reactions observed was significantly smaller in hypersensitivity than in the complement fixation test.


MiR-325 Promotes Oxaliplatin-Induced Cytotoxicity Against Colorectal Cancer Through the HSPA12B/PI3K/AKT/Bcl-2 Pathway.
Oxaliplatin is one of the most effective chemotherapeutic drugs used for the treatment of colorectal cancer (CRC). However, intervention that attenuates the resistance of oxaliplatin is still required in the treatment of CRC. To investigate the role of miR-325 in changing the oxaliplatin sensitivity to CRC cells. Expression of miR-325 in colorectal cancer tissues and cell lines was measured by using qRT-PCR analysis. Cytotoxicity of oxaliplatin to control or miR-325-overexpressed HT29 and SW480 cells was evaluated by CCK-8 assays. Luciferase reporter assay was used to confirm the regulation of miR-325 on HSPA12B. Flow cytometry was performed to detect the mitochondrial membrane potential and cell apoptosis. Expression of miR-325 was decreased in colorectal cancer tissues and cell lines. However, overexpression of miR-325 can decrease the 50% inhibiting concentration of oxaliplatin to colorectal cancer cell lines HT29 and SW480. Mechanically, we confirmed that miR-325 targeted HSPA12B in colorectal cancer. Therefore, overexpression of miR-325 inhibited the phosphorylation of PI3K and AKT and decreased the expression of Bcl-2 to promote the oxaliplatin-induced mitochondrial apoptosis in colorectal cancer. MiR-325 sensitizes the colorectal cancer cells to oxaliplatin-induced cytotoxicity through the HSPA12B/PI3K/AKT/Bcl-2 pathway.


Ecchymosis in critical coronavirus disease 2019 (COVID-19) patient in Tangerang, Indonesia: a case report.
We have read with great interest the articles regarding cutaneous manifestations in Coronavirus Disease 2019 (COVID-19) infection. Studies showed 20.4% of COVID-19 patients developed cutaneous manifestations. COVID-19 causes vascular endothelial injury, vasculitis and prothrombotic state which might be the underlying cause of hemorrhagic cutaneous manifestations, disseminated intravascular coagulation (DIC), and multiple organ failures. To date, hemorrhagic cutaneous manifestations reported are chilblain-like lesion, purpura to ecchymosis, livedo reticularis, and dry gangrene. We present a case of ecchymosis in COVID-19-positive ICU hospitalized patient.


[Formula: see text] modified AC for removing gaseous elemental mercury from flue gas.
Mercury pollution from coal-fired power plants has always been an environmental concern, and adsorption technology is an effective method for Hg0 removal. In this study, copper-iron binary metal sulfide modified activated carbon ([Formula: see text]) adsorbent was synthesized. The performance and mechanism of mercury removal were tested by adsorption experiments and characterization methods. The results showed that the mercury removal efficiency of [Formula: see text] under simulated flue gas (SFG) conditions at 150 °C could reach 91%, which was higher than [Formula: see text] and [Formula: see text]. Compared with [Formula: see text], the introduction of Fe increased the proportion of [Formula: see text], which was usually linked to [Formula: see text], resulting in the generation of more [Formula: see text](CuISI) species. Meanwhile, the generation of active sulfur sites such as [Formula: see text] was generated, which had a facilitative effect on the oxidation of Hg0. Stable [Formula: see text] was the predominant product on the adsorbent surface. The 2% sulfur loadings had significantly improved the sulfur resistance of conventional AC.


Successful post-term pregnancy in scared bicornuate uterus: case report.
Pregnancies in the bicornuate uterus are usually considered high-risk because of their association with poor reproductive outcomes such as recurrent pregnancy loss, cervical insufficiency, low birthweight, preterm birth, malpresentation, cesarean delivery, and uterine rupture. The objective of the report was to show that patients with a scared bicornuate uterus at post-term could have successful pregnancy outcomes. We are presenting a 28-year-old gravida III para II lady with a bicornuate scared uterus at post-term. She has a history of early third-trimester pregnancy loss, and her second pregnancy was delivered via cesarean section. In her first pregnancy, the patient's uterus was not identified as bicornuate. However, an ultrasound during her second pregnancy revealed that she had a bicornuate uterus, which led to the diagnosis. At term, the lady had a successful cesarean section. Lastly, for the current post-term pregnancy she had no prenatal check-up. Even though this, she was coming at the latent first stage of labor and an emergency cesarean section was done.Conclusion Successful outcomes could be achieved in patients with the bicornuate uterus at post-term gestation.


Norethandrolone produces temporary loss of the ability to escape from salt-retaining steroids.
Patients with diseases characterized by salt retention manifest a loss of the normal ability of healthy persons to escape from repeat injections of aldosterone or other salt-retaining steroids. This phenomenon may be a clue to the pathophysiological mechanisms of salt retention. Administration of norethandrolone to a subject who had demonstrated the ability to escape from the salt-retaining effect of corticosteroid administration temporarily and reversibly deleted his ability to escape. Thus norethandrolone administration provides the basis for a model system for exploring the mechanisms of escape (and therefore of salt retention).


Assessment of a Pharmaceutical Advertisement Analysis Module in a Drug Literature Evaluation Course.
Objective. To evaluate the impact of an educational module on students' self-efficacy when analyzing the content of promotional drug brochures (PDBs) and to assess the students' value of PDBs' as an educational tool. Methods. Third-year bachelor of pharmacy students participated in a one-hour lecture and a two-hour laboratory. Students completed a survey before and after participating in the module. Results. The module elicited a statistically significant change in students' self-efficacy beliefs regarding evaluating promotional drug brochures, while the average perceived value of promotional drug brochures did not change significantly after the module. Conclusion. A brief educational module can increase students' self-efficacy in evaluating the content of PDBs.


[Impact of glutamine of gut permeability and clinical prognosis on the aging patients undergoing gastric-intestinal operation].
To evaluate the impact of parenteral nutrition supplemented glutamine on aging patients undergoing gastric-intestinal operation. 30 patients above 60 years old undergoing gastric-intestinal operation, a randomized double-blind protocol was designed, divided into two groups, received impact isocaloric parenteral nutrition. The study group received alanyl-glutamine [0.5 g/(kg.d)]. To observe plasma amino acids profile, nitrogen balance, intestinal permeability and clinical prognosis, examine clinical chemistry variables and observe the adverse reactions in order to find out its safety. The patients in both groups were comparable prior to the operation. The plasma glutamine level of study group is higher than the control group, it's cumulative nitrogen balance values were prior to the control group, L/M ratio was lower than the control group. The complications related to infection was observed more in the control group. No adverse reaction was observed in both groups. Ala-Gln-supplemented PN improved nitrogen balance and maintained intestinal permeability, reduced complications.


Targeted silencing of the Aquaporin 2 gene of Rhipicephalus (Boophilus) microplus reduces tick fitness.
Ticks are blood-feeding arthropods that can affect human and animal health both directly by blood-feeding and indirectly by transmitting pathogens. The cattle tick Rhipicephalus (Boophilus) microplus is one of the most economically important ectoparasites of bovines worldwide and it is responsible for the transmission of the protozoan Babesia bovis, the etiological agent of bovine babesiosis. Aquaporins (AQPs) are water channel proteins implicated in physiological mechanisms of osmoregulation. Members of the AQP family are critical for blood-feeding arthropods considering the extreme osmoregulatory changes that occur during their feeding. We investigated the pattern of expression of a newly identified AQP2 gene of R. microplus (RmAQP2) in different tick tissues and stages. We also examined in vivo the biological implications of silencing expression of RmAQP2 silencing during tick feeding on either uninfected or B. bovis-infected cattle. In silico gene analyses were performed by multiple alignments of amino acid sequences and topology prediction. Levels of RmAQP2 transcripts in different tick tissues and stages were analyzed by reverse transcriptase quantitative PCR. Patterns of expression of RmAQP2 protein were investigated by immunoblots. Gene silencing was performed by RNA interference and in vivo functional analyses carried out by feeding ticks on either uninfected or B. bovis-infected cattle. RmAQP2 transcripts were found in unfed larvae, engorged nymphs, and salivary glands and guts of partially engorged females; however, of all tick tissues and stages examined, RmAQP2 protein was found only in salivary glands of partially engorged females. RmAQP2 silencing significantly reduced tick fitness and completely abrogated protein expression. The effect of RmAQP2 silencing on fitness was more pronounced in females fed on a B. bovis-infected calf than in ticks fed on an uninfected calf and none of their larval progeny survived. Collectively, considering the gene expression and tick fitness data, we conclude that RmAQP2 is critical for tick blood feeding and may be a suitable candidate target for the development of novel strategies to control R. microplus and tick-borne parasites.


Identification of Ehrlichia ruminantium proteins that activate cellular immune responses using a reverse vaccinology strategy.
Ehrlichia ruminantium is an obligate intracellular bacterial pathogen which causes heartwater, a serious tick-borne disease of ruminants throughout sub-Saharan Africa. The development of promising recombinant vaccines has been reported previously, but none has been as effective as immunisation with live organisms. In this study we have used reverse vaccinology to identify proteins that elicit an in vitro cellular immune response similar to that induced by intact E. ruminantium. The experimental strategy involved four successive steps: (i) in silico selection of the most likely vaccine candidate genes from the annotated genome; (ii) cloning and expression of the selected genes; (iii) in vitro screening of the expressed proteins for their ability to induce interferon-gamma (IFN-γ) production in E. ruminantium-immune lymphocytes; and (iv) further examination of the cytokine response profiles of those lymphocytes which tested positive for IFN-γ induction. Based on their overall cytokine induction profiles the recombinant proteins were divided into four distinct groups. Eleven recombinant proteins induced a cytokine profile that was similar to the recall immune response induced by immune peripheral blood mononuclear cells (PBMC) stimulated with intact E. ruminantium. This response comprised the upregulation of cytokines associated with adaptive cellular immune responses as well as innate immunity. A successful vaccine may therefore need to contain a combination of recombinant proteins which induce both immune pathways to ensure protection against heartwater.


Biochemical studies of six nitrogen-containing heterocycles in rat tissues.
Female rats were dosed orally with one-fifth the LD50 of either 1-nitrosopiperidine (a carcinogen), cyclohexylamine, piperidine, 4-carboxy-1-nitrosopiperidine, 4-cyclohexyl-1-nitrosopiperidine or 2,6-dimethyl-1-nitrosopiperidine at 21 and 4 hr before they were killed. The five noncarcinogenic compounds had no effects on any experimental variables examined [hepatic DNA damage, ornithine decarboxylase (ODC) activity, serum alanine aminotransferase (SGPT) activity, cytochrome P-450 and glutathione content]. After administration of 40 mg/kg of 1-nitrosopiperidine, marked hepatic DNA damage and a 3- to 7-fold increase in the activity of hepatic ODC were observed. 1-Nitrosopiperidine (120 mg/kg, 3/5 LD50) caused DNA damage in rat liver and esophagus but not in leukocytes. This higher dose of 1-nitrosopiperidine also increased hepatic ornithine decarboxylase activity by 9-fold. Thus, this hepatic biochemical assay system correctly identified the one carcinogen and the five noncarcinogens in this series of six nitrogen-containing heterocycles.


Mobile echoes in liver cysts: a form of range-ambiguity artifact.
Faint moving echoes are occasionally encountered in large hepatic cysts, as an example of range-ambiguity artifacts. The aim of this article is to describe the pattern of these intracystic mobile echoes, to analyze the mechanism of their formation, and to discuss options to clear them. We analyzed the size and location of the hepatic cysts, the movement of the artifactual echoes, and the relationship between pulse repetition frequency (PRF) and the depth of these intracystic mobile echoes in 10 cases. In three cases examination at a lower PRF was made to ascertain whether the artifactual echoes would disappear. Intracystic range-ambiguity echoes appeared when the heart was located distal to the hepatic cyst and these echoes moved according to cardiac motion. The depth of the intracystic artifacts changed according to the PRF and they disappeared at a low PRF. Intracystic range-ambiguity artifacts are caused by an erroneous display of the returning echoes from the heart. Knowledge of the mechanism and appearance of this artifact helps prevent misdiagnosis of internal echoes in large hepatic cysts. Observation at different PRFs is key to recognizing this artifact, and examination at lower PRFs should be done to confirm the artifactual nature of the echoes.


Treatment of mallet finger fractures by the extension-block K-wire technique.
We treated 22 patients with mallet finger fractures involving more than one-third of the articular surface by the extension-block K-wire technique. There were 18 men and four women with a mean age of 23 years (range, 14-34). The Wehbé and Schneider method was used to classify the fractures and the results were graded according to Crawford's criteria. All the fractures united, with an average healing time of 5.6 weeks (range, 4-7). At a mean follow-up of 25 months (range, 18-48) 21 mallet finger fractures had an excellent or good result. One patient had a fair result with a lag to extension of 20 degrees. We conclude that the extension-block K-wire technique is an effective method of treatment for displaced mallet finger fractures.


An Ultra-Thin MXene Film for Multimodal Sensing of Neuroelectrical Signals with Artifacts Removal.
Neuroelectrical signals transmitted onto the skin tend to decay to an extremely weak level, making them highly susceptible to interference from the environment and body movement. Meanwhile, for comprehensively understanding cognitive nerve conduction, multimodal sensing of neural signals, such as magnetic resonance imaging (MRI) and functional near-infrared spectroscopy (fNIRS), is highly required. Previous metal or polymer conductors cannot either provide a seamless on-skin feature for accurate sensing of neuroelectrical signals or be compatible with multimodal imaging techniques without opto- and magnet- artifacts. Herein, a ≈20 nm thick MXene film that is able to simultaneously detect electrophysiological signals and perform imaging by MRI and fNIRS with high fidelity is reported. The ultrathin film is made of crosslinked Ti3 C2 Tx film via poly (3,4-ethylene dioxythiophene): polystyrene sulfonate (PEDOT: PSS), showing a record high electroconductivity and transparency combination (11 000 S cm-1 @89%). Among them, PEDOT: PSS not only plays a cross-linking role to stabilize MXene film but also shortens the interlayer distance for effective charge transfer and high transparency. Thus, it can achieve a low interfacial impedance with skin or neural surfaces for accurate recording of electrophysiological signals with low motion artifacts. Besides, the high transparency originating from the ultrathin feature leads to good compatibility with fNIRS and MRI without optical and magnetic artifacts, enabling multimodal cognitive neural monitoring during prolonged use.


Macrophage-Specific NLRC5 Protects From Cardiac Remodeling Through Interaction With HSPA8.
Macrophages regulate inflammation and the process of tissue repair. Therefore, a better understanding of macrophages in the pathogenesis of heart failure is needed. In patients with hypertrophic cardiomyopathy, NLRC5 was significantly increased in circulating monocytes and cardiac macrophages. Myeloid-specific deletion of NLRC5 aggravated pressure overload-induced pathological cardiac remodeling and inflammation. Mechanistically, NLRC5 interacted with HSPA8 and suppressed NF-κB pathway in macrophages. The absence of NLRC5 in macrophages promoted the secretion of cytokines such as interleukin-6 (IL-6), which affected cardiomyocyte hypertrophy and cardiac fibroblast activation. Tocilizumab, an anti-IL-6 receptor antagonist, may be a novel therapeutic strategy for cardiac remodeling and chronic heart failure.


Self-rating and objective memory testing of normal and depressed elderly.
This study aimed to assess the relationships between the scores of subjective assessment (metamemory) and those of performance testing for memory, on the one hand, and the level of depression, on the other. A hundred and eighty elderly subjects (102 women and 78 men; mean age 65.7 years) were selected for the study. They showed neither intellectual impairment (as assessed through Mini Mental State test: MMS) nor neuropsychiatric symptoms. Each subject was administered the Randt Memory Test (RMT) for performance testing, the Sehulster Memory Scale (SMS) for the subjective assessment, and the Geriatric Depression Scale (GDS). A MULTCOVA analysis showed that both age and the depression level are negatively correlated with both the measures (Acquisition-Recall: AR; Delayed Memory: DM) of the RMT. The scores of the second (memory complaints) of the three sets of SMS were positively correlated with those of AR and DM indices. A Multivariate Regression Analysis showed that in males age and the depression level were significant regressors for both AR and MD scores while in females only the depression level was a significant regressor for AR and only age was a significant regressor for DM. Our results suggest that a) the relationships between the depression level and memory functioning are close, although not fully homogeneous in men and women; and b) that the scores in some areas of metamemory parallel, independently of the level of depression, the performance outcomes of memory functioning.


Behavioral alterations of zebrafish larvae after early embryonic exposure to ketamine.
Ketamine has been associated with pediatric risks that include neurocognitive impairment and long-term behavioral disorders. However, the neurobehavioral effects of ketamine exposure in early development remain uncertain. This study aimed to test stage- and dose-dependent effects of ketamine exposure on certain brain functions by evaluating alterations in locomotion, anxiety-like and avoidance behaviors, as well as socialization. Embryos were exposed to different concentrations of ketamine (0, 0.2, 0.4, and 0.8 mg mL-1) for 20 min during the 256-cell (2.5 h post fertilization-hpf), 50% epiboly (5.5 hpf), and 1-4 somites (10.5 hpf) stages. General exploratory activities, natural escape-like responses, and social interactions were analyzed under continuous light or under a moving light stimulus. A dose-dependent decrease in the overall mean speed was perceived in the embryos exposed during the 256-cell stage. These results were related to previously observed head and eye malformations, following ketamine exposure at this stage and may indicate possible neurobehavioral disorders when ketamine exposure is performed at this stage. Results also showed that ketamine exposure during the 50% epiboly and 1-4 somites stages induced a significant increment of the anxiety-like behavior and a decrease in avoidance behavior in all exposed groups. Overall, the results validate the neurodevelopmental risks of early-life exposure to ketamine.


Release of some endogenous trace amines from rat striatal slices in the presence and absence of a monoamine oxidase inhibitor.
The basal and 50 mM K+-stimulated release of m-tyramine (mTA), p-tyramine (pTA), tryptamine (TR) and phenylethylamine (PE) from striatal slices obtained from rats pretreated with a monoamine oxidase inhibitor (MAOI) was investigated. A K+-stimulated release of mTA and pTA was observed, but K+ did not stimulate either TR or PE release. The latter two amines, therefore, are unlikely to be conventional neurotransmitters in the rat striatum. The release of endogenous striatal pTA from control rats was also investigated. Veratridine stimulated endogenous pTA release, but 50 mM K+ did not. It is possible, therefore, that endogenous pTA can be released in a transmitter-like fashion.


Structure and evolution of insulins: implications for receptor binding.
Insulin is a member of a family of hormones, growth factors and neuropeptides which are found in both vertebrates and invertebrates. A common 'insulin fold' is probably adopted by all family members. Although the specificities of receptor binding are different, there is a possibility of co-evolution of polypeptides and their receptors.


The effect of ZIF-67 nanoparticles on the desulfurization performance of deep eutectic solvent based nanofluid system.
The nanoparticles of zeolitic imidazolate framework (ZIF-67) were synthesized and added to ethanolamine/deep eutectic solvent solution to form nanofluid system. The dynamic removal performance of prepared nanofluid system for hydrogen sulfide was investigated. For the system based on choline chloride and urea, the introduction of nanoparticles showed significant enhancement effect on the desulfurization performance. The optimal mass fraction of nanoparticles in nanofluid systems were identified as 0.1%. Besides, the experimental results showed that the prepared nanofluid systems have high regeneration performance, and the presence of moderate moisture is beneficial to the regeneration process. The absorbents and nanoparticles before and after absorption were characterized by Fourier transform infrared spectra, nuclear magnetic resonance, scanning electron microscope, energy dispersive spectrum, X-ray diffraction and X-ray photoelectron spectroscopy. The characterization results showed that the surface of nanoparticle was covered by CoS2 after absorption.


Cofactor specificity engineering of a long-chain secondary alcohol dehydrogenase from Micrococcus luteus for redox-neutral biotransformation of fatty acids.
Structure-based engineering of a NAD+-dependent secondary alcohol dehydrogenase from Micrococcus luteus led to a 1800-fold increase in catalytic efficiency for NADP+. Furthermore, the engineered enzymes (e.g., D37S/A38R/V39S/T15I) were successfully coupled to a NADPH-dependent Baeyer-Villiger monooxygenase from Pseudomonas putida KT2440 for redox-neutral biotransformations of C18 fatty acids into C9 chemicals.


[Relationship between the fecal mutagenicity and metal content, smoking habit and dietary intake].
The study was carried out to collect basic data on exposure of mucous cells of the large intestine to mutagens by investigating the genotoxicity of fecal samples from the urban population. Simultaneously, relations with food intake (food groups and nutritive components), lifestyle factors and fecal metal content was investigated. A total of 199 self-collected fecal samples from middle aged healthy volunteer living in urban areas of Osaka city were freeze-dried and ground in a mill. The mutagenicity of aqueous extracts of the ground samples was measured by the umu-test. Metal content of the feces was analysed by atomic absorption spectrophotometlly. On the day previous to the feces collection, the contents of meals were recorded and confirmed by interview with a dietitian. Quantification of nutritive components was carried out using the 4th edition of the Japan Food Standard Vomposition Table. Mutagenicity of feces was higher in males than in females, and in those aged 40-49 years than 50-69 years in males but not females. Large differences were found for content of 8 metals in the feces; concentrations were in the decreasing order of calcium, potassium, magnesium, sodium, zinc, iron, manganese and copper, the highest being 20.4 mg for calcium and the lowest was 53.5 micrograms for copper. Between the metal contents and mutagenicity, values for zinc and iron showed positive correlations and for sodium a negative correlation under S9(+) conditions. A weak but significant correlation was observed between the numbers of cigarettes smoked per day and S9 (+) mutagenicity. However we could not find any relation with food groups or nutritive components. It is well known that human feces contain many mutagens and carcinogens and that these can act directly on the mucous membrane of large intestine. Therefore, it is very important to estimate exposure levels. From the present data we can conclude that relations between mutation-activity of the feces and diet are complex. Their elucidation will require a large number of volunteers who have similar living conditions to obtain appropriate data.


Cell-free microRNA-21: biomarker for intracranial aneurysm rupture.
Deregulation of miRNA-21 expression has been reported to be associated with vascular smooth muscle behavior and cytoskeletal stability. This study is aimed to investigate the density of serum miRNA-21 in patients with different phases of intracranial aneurysms (IAs) and explore its warning function for IA rupture. A total of 16 in 200 IA patients were selected and categorized into 4 groups based on the phase of IA. Microarray study was carried out using serum miRNA and differentially expressed miRNAs were identified. Another 24 samples from a cohort of 360 patients were added and real-time polymerase chain reaction (RT-PCR) was performed on expanded sample size (n = 40) for miRNA-21 validation. Potential gene targets of miRNA-21 were screened out from Gene Ontology (GO) database and literatures. Microarray study identified 77 miRNAs with significantly different expression levels between experimental groups and the control group. RT-PCR assays validated significant downregulation of miRNA-21 in experimental groups, among which miRNA-21 expression level of daughter aneurysm group decreased the most. Bioinformatic analyses revealed that several target genes related with miRNA-21 may be involved in IA formation and rupture. This study suggested that miRNA-21 had a protective effect for intracranial vascular wall against remodeling and warning function for intracranial aneurysm rupture. Significant suppression of serum miRNA-21 in IA patients may provide diagnostic clues for aneurysm rupture and guide clinical intervention.


Public Perceptions around mHealth Applications during COVID-19 Pandemic: A Network and Sentiment Analysis of Tweets in Saudi Arabia.
A series of mitigation efforts were implemented in response to the COVID-19 pandemic in Saudi Arabia, including the development of mobile health applications (mHealth apps) for the public. Assessing the acceptability of mHealth apps among the public is crucial. This study aimed to use Twitter to understand public perceptions around the use of six Saudi mHealth apps used during COVID-19: "Sehha", "Mawid", "Sehhaty", "Tetamman", "Tawakkalna", and "Tabaud". We used two methodological approaches: network and sentiment analysis. We retrieved Twitter data using specific mHealth apps-related keywords. After including relevant tweets, our final mHealth app networks consisted of a total of 4995 Twitter users and 8666 conversational relationships. The largest networks in size (i.e., the number of users) and volume (i.e., the conversational relationships) among all were "Tawakkalna" followed by "Tabaud", and their conversations were led by diverse governmental accounts. In contrast, the four remaining mHealth networks were mainly led by the health sector and media. Our sentiment analysis approach included five classes and showed that most conversations were neutral, which included facts or information pieces and general inquires. For the automated sentiment classifier, we used Support Vector Machine with AraVec embeddings as it outperformed the other tested classifiers. The sentiment classifier showed an accuracy, precision, recall, and F1-score of 85%. Future studies can use social media and real-time analytics to improve mHealth apps' services and user experience, especially during health crises.


Novel metronidazole-chalcone conjugates with potential to counter drug resistance in Trichomonas vaginalis.
Trichomoniasis is the most prevalent, curable sexually transmitted disease (STD), which increases risk of viral STDs and HIV. However, drug resistance has been developed by some strains of Trichomonas vaginalis against Metronidazole (MTZ), the FDA approved drug against trichomoniasis. In the present study twenty two chalcone hybrids of metronidazole have been synthesized in a quest to get new molecules with higher potential against metronidazole-resistant T. vaginalis. All new hybrid molecules were found active against T. vaginalis with varying levels of activity against MTZ-susceptible and resistant strains. Eight compounds (4a, 4c, 4d, 4e, 4f, 4h, 4q and 4s) were found as active as the standard drug with an MIC of 1.56 μg/ml against MTZ-susceptible strain. However, compounds 4e, 4h and 4m were 4-times more active than MTZ against drug-resistant T. vaginalis, amongst which 4e and 4h were most promising against both susceptible and resistant strains.


Extensions and adaptations of existing medical information system in order to reduce social contacts during COVID-19 pandemic.
The main objective of this paper is the reduction of the COVID-19 pandemic spread by increasing the degree of social distancing by using and upgrading the existing Medical Information System (MIS). The existing MIS MEDIS.NET, currently used in the largest health center in the Balkans, has been adapted and further developed. During the adaptation of existing MIS MEDIS.NET 4 new and 9 existing modules were developed. A quick questionnaire for the smart triage of patients was also implemented. The adapted MIS successfully influenced the reduction of social contacts within the Health Center Nis. The need for the arrival of children and their parents to receive appropriate health certificates for the school enrolment is reduced. The therapy of chronic patients has been prolonged for 6 months via an electronic prescription. An online service for the communication between patients and the chosen physicians is provided. Possible social contacts and exposure to the viral environment of patients are reduced by making appointments in extended slots and at determined physical locations. Patients are notified per SMS or email about the availability of chosen and physician on duty. The social distancing of patients and physicians is also established by sending laboratory analyses per email or SMS. Keeping the central registry for COVID-19 is enabled throughout the country. The smart adaptation of MIS, and its collaboration with other state systems can significantly influence the reduction of social contacts and thus mitigate the consequences of COVID-19 pandemic.


The application of caprylic acid in downstream processing of monoclonal antibodies.
Caprylic acid (CA), a naturally occurring eight-carbon fatty acid, has long been used as albumin stabilizer, non-IgG fraction precipitant and bactericidal agent in pharmaceutical industry. The mechanisms through which CA achieves its effects have been correlated with the molecule's protein/lipid binding capacity conferred by its octyl moiety. This article, following an initial review of CA's historical applications, introduces CA's relatively new application in downstream processing of monoclonal antibodies (mAbs). By taking advantage of CA mediated impurity precipitation and virus inactivation, it might be possible to develop a two-column purification process in replacement of the standard three-column process without compromising product quality.


Monitoring of proteolytic enzyme activity using phase transition-based peptide arrays.
We have developed an assay using peptide arrays based on phase transition from the glass substrate to the liquid for monitoring quantitative protease activity in real-time. Peptide arrays were fabricated using a bifunctional cross-linker, N-[γ-maleimidobutyryloxy] sulfosuccinimide ester, and a substrate peptide containing two functional groups, cysteine and tetramethyl-6-carboxyrhodamine (TAMRA) on the C- and N-terminus, respectively. The phase transition-based peptide arrays were characterized by analyzing the substrate peptide cleaved from the solid substrate by matrix metalloproteinase-3 (MMP-3). We successfully used this assay to determine the quantitative proteolytic activity of MMP-3 in a dose-dependent manner. In addition, parameters including Michaelis constant (K(m)), maximum rate of enzymatic reaction (V(max)), and half maximal inhibitory concentration (IC(50)) were determined by analyzing the concentrations of substrate peptide cleaved by MMP-3. Therefore, this new assay has potential for the quantitative analysis of enzyme kinetics of protease and informs research developments in drug discovery utilizing kinetic studies.


Intrachoroidal cavitation in macular area of eyes with pathologic myopia.
To determine the incidence and characteristics of intrachoroidal cavitations in the macular area of eyes with high myopia. Prospective, noninterventional case series. We evaluated 56 eyes of 44 patients with pathologic myopia (myopic spherical equivalent >8 diopters) and with patchy chorioretinal atrophy using a swept-source optical coherence tomographic (OCT) system with a center wavelength of 1050 nm. We focused on the changes in the scleral curvature in the area of patchy atrophy. The relationship of the macular intrachoroidal cavitation and retinoschisis was also analyzed. Sixty-eight consecutive patients with pathologic myopia but without patchy atrophy were analyzed as controls. In 31 of 56 eyes (55.4%) with patchy atrophy, the swept-source OCT images showed that the sclera was bowed posteriorly in and around the patchy atrophy compared to neighboring sclera, whereas none of the 68 patients without patchy atrophy showed this finding. Macular intrachoroidal cavitation had OCT features similar to peripapillary intrachoroidal cavitation; the choroid in the macular intrachoroidal cavitation area appeared thickened and the retina was caved into the cavitation. There was a direct communication between the vitreous and intrachoroidal cavitation in 3 eyes. Retinoschisis was observed significantly more frequently in or around the patchy atrophy in eyes with macular intrachoroidal cavitation than in those without cavitation. These findings suggest that patchy atrophy affects the scleral contour within posterior staphyloma beyond the funduscopically identified patchy atrophy by macular intrachoroidal cavitation. Such deformation of sclera may facilitate the development of retinoschisis in and around the patchy atrophy.


Broncholithiasis From Histoplasmosis in a Pediatric Patient: Case Reports and Review of Literature.
Broncholithiasis is a calcification in the airway. Though broncholiths are not common, it is even more unusual in a pediatric patient. We present a 15-year-old immunocompetent pediatric patient who had a broncholith that was found and removed by bronchoscopy.


Influence of coculture with Sertoli cells on steroidogenesis in immature rat Leydig cells.
The hypothesis has been advanced that Sertoli cells produce one or more follicle-stimulating hormone (FSH)-dependent paracrine factors which stimulate Leydig cell maturation and steroidogenesis. In an attempt to identify these factors we studied the effect of coculture with Sertoli cells on the steroidogenic capacity of immature Leydig cells. It is demonstrated that coculture, during a period of 6 days, markedly increases the capacity of the Leydig cells to secrete C21-steroids (progesterone, 17 alpha-hydroxyprogesterone, 20 alpha-hydroxypregn-4-en-3-one) and C19-steroids (testosterone, androst-4-ene-3,17-dione) in response to stimulation with luteinizing hormone (LH). Pretreatment of the cocultures on days 4, 5 and 6 with low concentrations of gonadotropins further enhances the steroidogenic response to LH. This pretreatment results in an overall increase in steroid output. At low concentrations of Sertoli cells and when short incubation times are used, pretreatment with FSH is clearly more effective than pretreatment with LH. Pretreatment with gonadotropins also results in a disproportionate increase in C19 output caused by increased conversion of C21 precursors into C19-steroids. This effect is also observed in Leydig cell monocultures and is mainly due to LH action on Leydig cells. Finally pretreated cocultures display a selective increase in testosterone output. The latter effect is caused by FSH-dependent conversion of androstenedione into testosterone in Sertoli cells. Pretreated cocultures can be maintained for at least 28 days. During this entire period their basal steroid output increases. Using a two-chamber culture system it is demonstrated that direct cell-cell interactions are not required to observe the stimulatory effects of coculture. One or more diffusible factors are involved and continuous contact with these factors is required to maintain the effect. Immunoneutralization experiments using an insulin-like growth factor (IGF-I) antiserum show that IGF-I is an important permissive factor to maintain steroidogenesis in isolated Leydig cells and in cocultures. Under none of the conditions studied, however, does the antiserum neutralize the stimulatory effect of coculture. It is concluded that the stimulatory effects of coculture on the testosterone output of Leydig cells are complex and that important diffusible mediators still remain to be identified.


Component-resolved immunologic modifications, efficacy, and tolerance of latex sublingual immunotherapy in children.
As the frequency of natural rubber latex (NRL) allergy has increased, attempts have been made to diminish exposure in high-risk patients. Despite some good results, complete NRL avoidance was not possible, so latex immunotherapy was developed. To examine variations in immunologic parameters, clinical efficacy, and safety of NRL sublingual immunotherapy (SLIT). This prospective, observational, open, case-control study included 23 patients (18 patients receiving NRL SLIT and 5 controls). Skin prick, conjunctival provocation, and in-use tests with NRL, specific IgE and specific IgG4 to NRL, specific IgE to recombinant NRL allergens, and basophil activation test (BAT) with whole latex, natural, and recombinant allergens were performed before immunotherapy (T0) and at 6 (T1) and 12 months (T2) of treatment. Patients were sensitized to Hev b 5, Hev b 6.01, and Hev b 6.02 proteins, optimal for SLIT. Changes in specific IgE were not significant. Increases in specific IgG4 between T1 and T2 were larger in the active group. BAT determinations showed significant decreases in recombinant Hev b 6.01 and natural Hev b 6.02 in the active group at T1 but not at T2. Both groups had new sensitizations at T1 but not at T2. The active group had significant increases in the response threshold in the in vivo tests at T1 and T2. Adverse effects were limited to local reactions. NRL SLIT is effective and safe in children with latex allergy. Our results suggest that specific IgE determinations and BAT measurements to natural and recombinant latex allergens may allow obtaining an allergen-based diagnosis to help determine specific immunotherapy.


Nuclear localization of huntingtin during spermatogenesis.
Huntingtin is a ubiquitous cytoplasmic protein. Mutant huntingtin causes Huntington's disease and its intranuclear inclusion is associated with cytotoxicity. Nuclear localization of normal huntingtin is detected in the oocyte up to 2.5 days post coitum. Therefore, huntingtin is expected to reside in the nucleus even before fertilization. The present study determined normal huntingtin distribution during spermatogenesis. Testicles from an adult male Sprague-Dawley rat were stained with anti-huntingtin antibody and nuclear counterstaining was performed with 4',6-diamidino-2-phenylindole. Concerning nuclear localization, huntingtin was detected in the spermatids, whereas predominant cytoplasmic localization of it was evident in the spermatogonia. Between the primary and secondary spermatocytes, huntingtin appeared to be delocalized in the nuclei when meiosis occurred. The findings provide evidence that normal huntingtin is transported to the nuclear compartment during the meiotic stage of spermatogenesis.


High-performance liquid chromatographic analysis of bioactive triterpenes in Perilla frutescens.
Perilla frutescens (L.) Britt. (Lamiaceae), a famous traditional Chinese medicine, has been used for the treatment of various diseases. To evaluate the quality of P. frutescens, a simple, rapid and accurate high-performance liquid chromatography (HPLC) method was developed for the assessment of three bioactive triterpene acids: tormentic acid (TA), oleanolic acid (OA) and ursolic acid (UA). The HPLC system used an Spherisob octadecylsilyl silica (ODS) column with acetonitrile and aqueous H(3)PO(4) as the mobile phase and detection at 206 nm. The method was precise with relative standard deviations for these three constituents that ranged between 0.6-1.5% (intraday) and 0.7-2.6% (interday). The content of these three phytochemicals in the leaves of P. frutescens growing at eight different locations of China was determined to establish the effectiveness of the method.


Acute lymphoblastic leukemia with a unique translocation in an adolescent boy.
We report a case of an adolescent boy with acute lymphoblastic leukemia whose blasts had three chromosomal abnormalities: trisomy 8, a t(5;15), and an extra "marker" chromosome. The patient presented with huge hepatosplenomegaly and pancytopenia. The response to treatment (ALL BFM 90 protocol) was very rapid, and the patient is in complete remission 1 year after diagnosis.


Platelet-to-lymphocyte ratio and prognosis in STEMI: A meta-analysis.
Platelet-to-lymphocyte ratio (PLR) is a haematological index which reflects increased level of inflammation and thrombosis. We aimed to summarize the potential prognostic role of PLR for the in-hospital and long-term outcomes in ST-segment elevation myocardial infarction (STEMI) patients treated with primary percutaneous coronary intervention (pPCI) in a meta-analysis. Relevant cohort studies were identified by search the PubMed, Cochrane's Library and Embase databases. A random-effect model was applied to pool the results. In-hospital and long-term outcomes were compared between patients with higher and lower preprocedural PLR. Eleven cohorts with 12 619 patients were included. Pooled results showed that higher preprocedural PLR was independently associated with increased risk of in-hospital major adverse cardiovascular events (MACE, risk ratio [RR]: 1.76, 95% confidence interval [CI]: 1.39 to 2.22, P < .001; I2 = 49%), cardiac mortality (RR: 1.91, 95% CI: 1.18 to 3.09, P = .009; I2 = 0), all-cause mortality (RR: 2.14, 95% CI: 1.52 to 3.01, P < .001, I2 = 24%) and no reflow after pPCI (RR: 2.22, 95% CI: 1.70 to 2.90, P < .001, I2 = 59%). Moreover, higher preprocedural PLR was associated with increased risk of MACE (RR: 1.60, 95% CI: 1.25 to 2.03, I2 = 57%, P < .001) and all-cause mortality (RR: 2.36, 95% CI: 1.53 to 3.66, I2 = 78%, P < .001) during long-term follow-up of up to 82 months after discharge. Higher PLR predicts poor in-hospital and long-term prognosis in STEMI patients after pPCI.


PCDD/Fs traceability during triclosan electrochemical oxidation.
5-Chloro-2-(2,4-dichlorophenoxy)phenol (TCS) is a persistent organic pollutant (POP) widely used in different consumer goods. Its recalcitrant nature demands the application of effective remediation technologies in order to avoid the negative environmental impact associated to the discharge of contaminated waters. Although advanced oxidation technologies have been considered the best alternative to destroy bio-recalcitrant compounds, the likely formation of high toxicity byproducts must be analysed before large-scale deployment. In this work, we aim to trace the presence of chlorinated compounds during the electro-oxidation of aqueous TCS samples. First, we analyze the influence of the initial concentration of TCS on the toxicity of the oxidation medium expressed by the International-Toxicity Equivalency Factor (I-TEF); second, we have detected the formation of intermediate organo-chlorinated compounds by GC-MS supported by HPLC and finally, we have quantified the concentration of highly-polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) by HRGC-HRMS within the oxidation treatment. In those samples where TCS had been completely degraded the concentration of PCDD/Fs showed a high increase, especially when NaCl was used as electrolyte, with the initial concentration of TCS. Under these conditions the I-TEF achieved values up to 3.8 × 102 pg L-1.


Changes in knee cartilage volume and serum COMP concentration after running exercise.
Investigate the relationship between running induced joint loading at the knee, changes in cartilage volume and serum cartilage oligomeric matrix protein (COMP) concentration. Serum COMP levels and knee cartilage volumes of experienced runners were tested before and after running. Joint loading was determined using a biomechanical model of the lower extremity. To date no biomechanical rationale has been identified to explain the role of mechanical load in the aetiology of running injuries. Blood samples and magnetic resonance imaging scans were taken before and following a 1h training run. Knee cartilage volume and serum COMP concentration were determined. Individual knee joint loading parameters were calculated from positional data and ground reaction forces. Electromyography was employed to quantify activity of main muscle groups crossing the knee joint. Changes in cartilage volume and COMP showed significant correlations. Net joint forces did not explain the differences in cartilage changes. Multiple regression revealed that resting COMP, COMP change after exercise and the time of co-activation of flexor and extensor muscles explain the variance of cartilage volume changes. Muscular co-activation was the main mechanical parameter related to cartilage changes. The current investigation elucidates the interaction of factors related to cartilage degeneration on an individual basis. Applications to altered loading conditions such as equipment or training methods offer an auspicious way of quantifying effects of interventions.


Study on distribution of mycotoxins in cocoa beans.
Mycotoxins are not homogeneously distributed in foods which come in naturally small units, such as pistachios and peanuts, and may instead be extremely inhomogeneously distributed due to the occurrence of so-called hot spots. Tests conducted on pistachios, for example, show that a mouldy kernel can be so strongly contaminated with mycotoxins that it has a significant impact on the contamination profile of several thousand kernels. This makes a representative sampling of such foodstuffs very important but also a very difficult task. Whether cocoa beans also have a tendency to form so-called mycotoxin hot spots is hitherto unknown.A miniaturised analysis method was used in tests made on several independent batches of cocoa beans and although these tests showed that the mycotoxins ochratoxin A and the aflatoxins are not homogeneously distributed in cocoa, the tested batches revealed no real hot spots.


Elucidation of Gut Microbiota in Mud Crab Scylla paramamosain Challenged to WSSV and Aeromonas hydrophila.
Mud crab Scylla paramamosain (S. paramamosain) is an economically important marine crab species around the world. White spot syndrome virus (WSSV) and Aeromonas hydrophila (AH) are pathogens during mud crab mariculture. It has been reported that gut microbiota possessed a great impact on the host development, nutrition, immunity, and disease resistance. However, little information was known about the impacts of WSSV or AH infection on the structure, composition, and function of the gut microbiotain of mud crabs. In this study, the gut microbiota of mud crabs infected with A. hydrophila and WSSV were characterized. The results showed that the composition and bacteria correlation of the gut microbiota were significantly decreased. During A. hydrophila infection, the pathogens played a major regulatory role in host. While in the mud crabs infected with WSSV, many beneficial strains had a great impact on the host expect for the pathogens. Therefore, our study revealed the effect of pathogens infection on gut microbiota of mud crabs and clarified the difference between viral infection and bacterial infection.


A Phase II Study of Regorafenib With a Lower Starting Dose in Patients With Metastatic Colorectal Cancer: Exposure-Toxicity Analysis of Unbound Regorafenib and Its Active Metabolites (RESET Trial).
Regorafenib demonstrated survival benefits as salvage therapy for patients with metastatic colorectal cancer. However, severe toxicities frequently occurred early in the treatment with the standard dose (160 mg/day), resulting in a dose reduction or interruption. To improve the tolerability and maintain sufficient efficacy, we conducted a phase II study of regorafenib with a lower starting dose (120 mg/day). Regorafenib was initiated at 120 mg/day, and the dosage was increased to 160 mg/day on day 15 of the first cycle for patients who had met the dose escalation criteria. The primary endpoint was the disease control rate (DCR). The pharmacokinetics of the total and unbound regorafenib and its active metabolites (M2, M5) were assessed. A total of 70 patients were enrolled from September 2016 to December 2017. Only 6 patients achieved dose escalation to 160 mg on day 15 as planned. For the 68 evaluable patients, the DCR was 32.4% (95% confidence interval, 21.5%-44.8%), which was less than the threshold (30%) of our statistical hypothesis. The serum concentrations of total regorafenib for patients whose dose was escalated to 160 mg/day were significantly lower than those of the patients whose dose was not escalated (median, 3978 vs. 7244 nM; P = .027). The serum unbound concentrations of the sum of regorafenib and the active metabolites correlated significantly with the maximum grade of regorafenib-related symptomatic adverse events in the first cycle (11,138 vs. 19,096 pM; P = .035). Regorafenib with a low starting dose of 120 mg/day did not achieve the expected DCR. A relationship of unbound exposure with toxicity was found.


A delicate interplay of structure, dynamics, and thermodynamics for function: a high pressure NMR study of outer surface protein A.
Outer surface protein A (OspA) is a crucial protein in the infection of Borrelia burgdorferi causing Lyme disease. We studied conformational fluctuations of OspA with high-pressure (15)N/(1)H two-dimensional NMR along with high-pressure fluorescence spectroscopy. We found evidence within folded, native OspA for rapid local fluctuations of the polypeptide backbone in the nonglobular single layer β-sheet connecting the N- and C-terminal domains with τ << ms, which may give the two domains certain independence in mobility and thermodynamic stability. Furthermore, we found that folded, native OspA is in equilibrium (τ >> ms) with a minor conformer I, which is almost fully disordered and hydrated for the entire C-terminal part of the polypeptide chain from β8 to the C-terminus. Conformer I is characterized with ΔG(0) = 32 ± 9 kJ/mol and ΔV(0) = -140 ± 40 mL/mol, populating only ∼0.001% at 40°C at 0.1 MPa, pH 5.9. Because in the folded conformer the receptor binding epitope of OspA is buried in the C-terminal domain, its transition into conformer I under in vivo conditions may be critical for the infection of B. burgdorferi. The formation and stability of the peculiar conformer I are apparently supported by a large packing defect or cavity located in the C-terminal domain.


c-Jun N-terminal kinase mediates microtubule-depolymerizing agent-induced microtubule depolymerization and G2/M arrest in MCF-7 breast cancer cells.
Microtubule-binding agents (MBAs) form one of the most important anticancer-drug families, but their molecular mechanisms are poorly understood. MBAs such as paclitaxel (PTX) stabilize microtubules, whereas XRP44X (a novel pyrazole) and combretastatins A4 (CA4) destabilize microtubules. These two different types of MBAs have potent antitumor activity. Comparisons of their effects on signal transduction and cellular responses will help uncover the molecular mechanism by which MBAs affect tumor cells. We used MCF-7 cells to compare the effects of the three MBAs on the cytoskeleton, cell cycle distribution, and activation of the three major mitogen-activated protein kinase (MAPK) signaling cascades [extracellular signal-related kinases, c-Jun N-terminal kinase (JNK), and p38 MAPK] using pharmacological inhibitors. The G2/M phase arrest was induced following polymerization of microtubules by PTX and depolymerization by XRP44X and CA4. The three major MAPKs were rapidly activated by XRP44X, and extracellular signal-related kinases and p38 by PTX, whereas JNK did not quickly respond to PTX. Pharmacological inhibitors indicated that activation of JNK is principally required for XRP44X- and CA4-induced microtubule depolymerization and G2/M phase arrest. Our results suggest that early phosphorylation of JNK is a specific mechanism involved in microtubule depolymerization by certain MBAs.


Neuropathological analysis in spinal muscular atrophy type II.
We performed a neuropathological analysis, including in situ nick end labeling (ISEL) and immunohistochemistry, of two cases of clinicogenetically confirmed infantile spinal muscular atrophy (SMA) type II. Both cases showed severe reduction of the motor neurons and gliosis in the spinal cord and brain stem, although the occurrences of central chromatolysis and ballooned neurons were not frequent. Clark's and lateral thalamic nuclei, which are usually altered in SMA type I, were spared, whereas Betz cells in the precentral gyrus and large myelinated fibers in the lateral funiculus were reduced in number. Regarding apoptosis, only the younger case demonstrated a few ISEL-positive nuclei in the dorsal horn, with reduced Bcl-x expression level in the Purkinje cells. Unlike SMA type I, the expression of neurofilaments was not disturbed and the reduction in synaptophysin expression level in the anterior horn was mild. An oxidative stress-related product was deposited in atrophic motor neurons in the spinal cord, and neurons with nuclei immunoreactive for 8-hydroxy-2'-deoxyguanosine were found in the lateral thalamus. In contrast, the expression of glial glutamate transporters was not altered. These data suggest that oxidative stress and, to a lesser extent, apoptotic cell death, but not disturbed neurofilament metabolism or excitotoxicity, may be involved in neurodegeneration in SMA type II.


Long-term outcome after urinary diversion using the ileocecal segment in children and adolescents: Complications of the efferent segment.
Long-term outcomes are of special concern in children after urinary diversion. In a single institution study we evaluated retrospectively the long-term outcomes of urinary diversion in children, in whom the ileocecal segment had been used, in respect to complications of the efferent segment. The Mainz pouch was used in 107 children for continent urinary diversion. Indications were neurogenic bladder (53%, 57/107), exstrophy-epispadias complex (25%, 27/107), malignancy (13%, 14/107), and others (9%, 9/107). Continent cutaneous diversion was performed in 95 patients, and 12 patients received bladder augmentation/substitution with a continent cutaneous stoma. As efferent segment, we used the in situ submucosally embedded appendix in 55/107, and an intussuscepted ileal nipple valve in 45/107 patients, other in seven patients. Complication rates and degree of satisfaction with urinary diversion were evaluated by assessing medical records and using follow-up questionnaires. The median follow-up time was 15.9 years (0.4-27.6 years). There was a significant difference between the stenosis rates of appendical stoma and the intussuscepted ileal nipple valve. In general, 38% of patients (41/107) with a continent cutaneous stoma developed a stoma stenosis at the skin level, with 41% (17/41) of those having recurrent stenoses (2-6×). Of the patients with an appendix stoma, 49% (27/55) developed a stenosis, compared with 20% (9/45) of those with an intussuscepted ileal nipple valve (p < 0.001). In an attempt to reduce the incidence of stenosis, we implemented use of the ACE stopper during the last 2 years, but whether this is successful is subject to further investigations. For surgical treatment of stoma stenosis, reanastomosis of the efferent segment to the skin (n = 37, re-stenosis rate 37%, [14/37]) was superior to endoscopic scar incision (n = 44, re-stenosis rate 59% [26/44], OR 2.4). The overall continence rate was 85% (91/107). At the latest follow-up, 89% (49/55) of patients with an appendix stoma and 82% (37/45) of those with an intussuscepted ileal nipple valve were completely continent. The in situ submucosally embedded appendix and the intussuscepted ileal nipple valve are reliable continence mechanisms in the long-term median follow-up of nearly 16 years. The stomal stenosis rate is approximately double for the submucosal appendix, likely because of its smaller diameter in comparison with the ileal valve. Although we encountered a high overall rate of complications, the satisfaction rate with the diversion was high (99% [88/89] satisfied or very satisfied patients).


Oxidized low-density lipoprotein depletes PKCalpha and attenuates reactive oxygen species formation in monocytes/macrophages.
Preexposure of macrophages to oxidized low-density lipoprotein (oxLDL) attenuates formation of reactive oxygen species (ROS) upon stimulation with phorbol 12-myristate 13-acetate (PMA) or acute exposure to oxLDL. We examined whether attenuation of the oxidative burst is attributed to down-regulation of protein kinase C alpha (PKCalpha). Acute exposure of a mouse macrophage cell line (RAW 264.7) to both PMA and oxLDL provoked ROS generation that was blocked by the PKCalpha/beta1 inhibitor Go 6967. However, in RAW 264.7 macrophages preincubated with oxLDL, ROS formation in response to stimulation with oxLDL or PMA was reduced. Attenuated ROS production correlated with down-regulation of the amount of PKCalpha protein in a time-dependent manner and was maximal at 8 h and concentrations of 50-100 microg/ml oxLDL. PMA, a well-established PKCalpha activator, inhibited ROS formation as well when preincubated for 8 to 16 h. In cells stably overexpressing PKCalpha-EGFP, we noticed that ROS formation remained intact upon pre-exposure of these cells to oxLDL. Moreover, in these cells endogenous but not overexpressed PKCalpha-EGFP disappeared, further substantiating the importance of PKCalpha in stimulating ROS production. In addition, we noticed a concentration-dependent ability of oxLDL to halt ROS production. Whereas 10 microg/ml oxLDL was insufficient in attenuating ROS formation over an 8-h incubation period in RAW 264.7 cells, 50 microg/ml oxLDL impaired ROS generation. These results indicate that attenuation of the oxidative burst in oxLDL-pretreated macrophages is closely associated with down-regulation of PKCalpha, which is elicited in a dose- and time-dependent manner.


Assessment of an automated approach for variant interpretation in screening for monogenic disorders: A single-center study.
Automation has been introduced into variant interpretation, but it is not known how automated variant interpretation performs on a stand-alone basis. The purpose of this study was to evaluate a fully automated computerized approach. We reviewed all variants encountered in a set of carrier screening panels over a 1-year interval. Observed variants with high-confidence ClinVar interpretations were included in the analysis; those without high-confidence ClinVar entries were excluded. Discrepancy rates between automated interpretations and high-confidence ClinVar entries were analyzed. Of the variants interpreted as positive (likely pathogenic or pathogenic) based on ClinVar information, 22.6% were classified as negative (variants of uncertain significance, likely benign or benign) variants by the automated method. Of the ClinVar negative variants, 1.7% were classified as positive by the automated software. On a per-case basis, which accounts for variant frequency, 63.4% of cases with a ClinVar high-confidence positive variant were classified as negative by the automated method. While automation in genetic variant interpretation holds promise, there is still a need for manual review of the output. Additional validation of automated variant interpretation methods should be conducted.


Neuromuscular comorbidity, heart failure, and atrial fibrillation as prognostic factors in left ventricular hypertrabeculation/noncompaction.
There is some controversy concerning the prognosis of patients with left ventricular hypertrabeculation/noncompaction (LVHT). LVHT is frequently associated with neuromuscular disorders (NMDs). The aim of this study was to assess cardiac and neurological findings as predictors of mortality in patients with LVHT. The study included patients with LVHT diagnosed between June 1995 and January 2014 in one echocardiographic laboratory. They underwent a baseline cardiologic examination and were invited for a neurological examination. Between January and February 2014, their survival status was assessed. LVHT was diagnosed in 220 patients (68 female, aged 52 ± 17 years) with a prevalence of 0.35 %/year. During a follow-up of 72 ± 61 months, 65 patients died. The mortality was 5 %/year. A neurological investigation was performed on 173 patients (79 %) and revealed specific NMDs in 31 (14 %), NMD of unknown etiology in 103 (47 %), and normal findings in 39 (18 %) patients. In multivariate analysis, the predictors of mortality were increased age (p = 0.0001), presence of a specific NMD (p = 0.0062) or NMD of unknown etiology (p = 0.0062), heart failure NYHA III (p = 0.0396), atrial fibrillation (p = 0.0022), and sinus tachycardia (p = 0.0395). LVHT patients should undergo systematic neurological examinations. Whether an optimal therapy of heart failure and atrial fibrillation will improve the prognosis of LVHT patients needs to be addressed in further studies.


Conformations of Three Types of Ultra-Long-Chain Fatty Acids in Multicomponent Lipid Bilayers.
Ultra-long-chain fatty acids (ULCFAs) are biosynthesized in certain types of tissues, but their biological roles remain unknown. Here, we report how the conformation of ULCFAs depends on the length and unsaturated-bond ratio of the ultra-long chains and the composition of the host bilayer membrane using molecular dynamics simulations. The ultra-long chain of ULCFAs flips between the two leaflets and fluctuates among three conformations: elongated, L-shaped, and turned. Furthermore, we found that the saturated ultra-long chain exhibited an elongated conformation more frequently than the unsaturated chain. In addition, the truncation of the ultra-long chain at C26 had little effect on the remaining ULCFAs. ULCFAs respond to lipid-density differences in the two leaflets, and the ratio of the elongated and turned conformations changed to reduce this difference. However, in cholesterol-containing membranes, ULCFAs exhibit no density difference after the flip-flop of cholesterol removes the difference.


Analysis of structural differences between Ly-5 molecules of T- and B-cells.
A group of closely related high mol. wt (Mr) membrane glycoproteins is expressed with varying Mr on different subpopulations of lymphocytes, but the different Mr forms share the Ly-5 and T200 antigenic determinants. The Ly-5 molecule expressed by thymocytes has an Mr of 175,000 (175ly5). The antigenically related molecule on B-cells has an Mr of 210,000 (210ly5). It is not known whether the variations in size are due to differences in the polypeptide chain, post-translational modifications such as glycosylation, or both. In this report we examine the glycosylation of 175ly5 and 210ly5 to determine whether differences in carbohydrate moieties may account for the different Mr of these two Ly-5 species. Pronase digestion and alkaline borohydride treatment of these molecules labeled in the terminal galactose residues revealed that 210ly5 molecules have a more complex oligosaccharide pattern than 175ly5 molecules. While Ly-5 oligosaccharides from a T-cell tumor line were very similar to those of normal thymocytes, the pattern of Ly-5 carbohydrates from a B-cell tumor were somewhat different than those from normal B-cells. This report also presents evidence for O-linked sugars on Ly-5 molecules.


Structure-based investigation of fluorogenic Pepper aptamer.
Pepper fluorescent RNAs are a recently reported bright, stable and multicolor fluorogenic aptamer tag that enable imaging of diverse RNAs in live cells. To investigate the molecular basis of the superior properties of Pepper, we determined the structures of complexes of Pepper aptamer bound with its cognate HBC or HBC-like fluorophores at high resolution by X-ray crystallography. The Pepper aptamer folds in a monomeric non-G-quadruplex tuning-fork-like architecture composed of a helix and one protruded junction region. The near-planar fluorophore molecule intercalates in the middle of the structure and is sandwiched between one non-G-quadruplex base quadruple and one noncanonical G·U wobble helical base pair. In addition, structure-based mutational analysis is evaluated by in vitro and live-cell fluorogenic detection. Taken together, our research provides a structural basis for demystifying the fluorescence activation mechanism of Pepper aptamer and for further improvement of its future application in RNA visualization.


An omics data analysis method based on feature linear relationship and graph convolutional network.
Biological networks are known to be highly modular, and the dysfunction of network modules may cause diseases. Defining the key modules from the omics data and establishing the classification model is helpful in promoting the research of disease diagnosis and prognosis. However, for applying modules in downstream analysis such as disease states discrimination, most methods only utilize the node information, and ignore the node interactions or topological information, which may lead to false positives and limit the model performance. In this study, we propose an omics data analysis method based on feature linear relationship and graph convolutional network (LCNet). In LCNet, we adopt a way of applying the difference of feature linear relationships during disease development to characterize physiological and pathological changes and construct the differential linear relation network, which is simple and interpretable from the perspective of feature linear relationship. A greedy strategy is developed for searching the highly interactive modules with a strong discrimination ability. To fully utilize the information of the detected modules, the personalized sub-graphs for each sample based on the modules are defined, and the graph convolutional network (GCN) classifiers are trained to predict the sample labels. The experimental results on public datasets show the superiority of LCNet in classification performance. For Breast Cancer metabolic data, the identified metabolites by LCNet involve important pathways. Thus, LCNet can identify the module biomarkers by feature linear relationship and a greedy strategy, and label samples by personalized sub-graphs and GCN. It provides a new manner of utilizing node (molecule) information and topological information in the defined modules for better disease classification.


The evolution of neocortex in primates.
We can learn about the evolution of neocortex in primates through comparative studies of cortical organization in primates and those mammals that are the closest living relatives of primates, in conjunction with brain features revealed by the skull endocasts of fossil archaic primates. Such studies suggest that early primates had acquired a number of features of neocortex that now distinguish modern primates. Most notably, early primates had an array of new visual areas, and those visual areas widely shared with other mammals had been modified. Posterior parietal cortex was greatly expanded with sensorimotor modules for reaching, grasping, and personal defense. Motor cortex had become more specialized for hand use, and the functions of primary motor cortex were enhanced by the addition and development of premotor and cingulate motor areas. Cortical architecture became more varied, and cortical neuron populations became denser overall than in nonprimate ancestors. Primary visual cortex had the densest population of neurons, and this became more pronounced in the anthropoid radiation. Within the primate clade, considerable variability in cortical size, numbers of areas, and architecture evolved.