Exploiting the biological response of two Serratia fonticola strains to the critical metals, gallium and indium.
The use of microorganisms that allows the recovery of critical high-tech elements such as gallium (Ga) and indium (In) has been considered an excellent eco-strategy. In this perspective, it is relevant to understand the strategies of Ga and In resistant strains to cope with these critical metals. This study aimed to explore the effect of these metals on two Ga/In resistant strains and to scrutinize the biological processes behind the oxidative stress in response to exposure to these critical metals. Two strains of Serratia fonticola, A3242 and B2A1Ga1, with high resistance to Ga and In, were submitted to metal stress and their protein profiles showed an overexpressed Superoxide Dismutase (SOD) in presence of In. Results of inhibitor-protein native gel incubations identified the overexpressed enzyme as a Fe-SOD. Both strains exhibited a huge increase of oxidative stress when exposed to indium, visible by an extreme high amount of reactive oxygen species (ROS) production. The toxicity induced by indium triggered biological mechanisms of stress control namely, the decrease in reduced glutathione/total glutathione levels and an increase in the SOD activity. The effect of gallium in cells was not so boisterous, visible only by the decrease of reduced glutathione levels. Analysis of the cellular metabolic viability revealed that each strain was affected differently by the critical metals, which could be related to the distinct metal uptakes. Strain A3242 accumulated more Ga and In in comparison to strain B2A1Ga1, and showed lower metabolic activity. Understanding the biological response of the two metal resistant strains of S. fonticola to stress induced by Ga and In will tackle the current gap of information related with bacteria-critical metals interactions.


Classical oncogenes and tumor suppressor genes: a comparative genomics perspective.
We have curated a reference set of cancer- related genes and reanalyzed their sequences in the light of molecular information and resources that have become available since they were first cloned. Homology studies were carried out for human oncogenes and tumor suppressors, compared with the complete proteome of the nematode, Caenorhabditis elegans, and partial proteomes of mouse and rat and the fruit fly, Drosophila melanogaster. Our results demonstrate that simple, semi-automated bioinformatics approaches to identifying putative functionally equivalent gene products in different organisms may often be misleading. An electronic supplement to this article provides an integrated view of our comparative genomics analysis as well as mapping data, physical cDNA resources and links to published literature and reviews, thus creating a "window" into the genomes of humans and other organisms for cancer biology.


Patterns of experience with verbs affect long-term cumulative structural priming.
Recent studies of structural priming have demonstrated that although there is evidence of verb-based effects in short-term priming (e.g., stronger priming when verbs are repeated between prime and target sentences), such effects are absent in long-term priming. We present evidence that verb-based effects can be observed in long-term priming situations. This result has important implications for theoretical accounts of the mechanisms that give rise to structural priming and other such adaptation effects in language production.


Homogeneous liquid-liquid microextraction via flotation assistance for rapid and efficient determination of polycyclic aromatic hydrocarbons in water samples.
In this work, a rapid, simple and efficient homogeneous liquid-liquid microextraction via flotation assistance (HLLME-FA) method was developed based on applying low density organic solvents without no centrifugation. For the first time, a special extraction cell was designed to facilitate collection of the low-density solvent extraction in the determination of four polycyclic aromatic hydrocarbons (PAHs) in water samples followed by gas chromatography-flame ionization detector (GC-FID). The effect of different variables on the extraction efficiency was studied simultaneously using experimental design. The variables of interest in the HLLME-FA were selected as extraction and homogeneous solvent volumes, ionic strength and extraction time. Response surface methodology (RSM) was applied to investigate the optimum conditions of all the variables. Using optimized variables in the extraction process, for all target PAHs, the detection limits, the precisions and the linearity of the method were found in the range of 14-41 μg L(-1), 3.7-10.3% (RSD, n=3) and 50-1000 μg L(-1), respectively. The proposed method has been successfully applied to the analysis of four target PAHs in the water samples, and satisfactory results were obtained.


Use of transoral carbon dioxide laser for supraglottic web from caustic ingestion in a 13 year old.
A 13 year old male was referred to the Department of Otorhinolaryngology due to unsuccessful decannulation. At 2 years of age, patient accidentally ingested a caustic liquid alkali and underwent emergency tracheostomy, exploratory laparotomy, and tube gastrectomy. 11 years after, patient was seen at the outpatient department for decannulation. On nasopharyngolaryngoscopy, a supraglottic stenosis with a central 1-mm opening was visualized. Patient underwent transoral carbon dioxide laser excision of supraglottic stenosis. Fused aryepiglottic folds were released from the epiglottis. Patient was discharged well with no immediate complications and was successfully decannulated 1 month post operation with good voice outcome. No recurrence of stenosis at one year post operation. Accidental caustic ingestion commonly occurs in the pediatric age group. A frequent complication of accidental caustic ingestion is strictures. The use of transoral carbon dioxide laser in the surgical treatment of supraglottic stenosis is a suitable option for pediatric patients as this offer several advantages such as less post operative edema, good intraoperative control of hemorrhage, few post-operative complications and is less invasive compared to open laryngeal procedures. This case exhibits the importance of close monitoring for pediatric patients after caustic ingestion in order to diagnose complications earlier.


Cerebral edema associated with diabetic ketoacidosis: Two case reports.
Diabetic ketoacidosis (DKA) is associated with a high mortality rate, especially if cerebral edema develops during the disease course. It is rarer and more severe in adults than in children. We present cases of two patients with cerebral edema-related DKA. The first patient was a 38-year-old man with diabetes mellitus who presented with DKA-related disturbed consciousness. Although glycemic correction was performed slowly, he showed pupil dilation 11 h later. He underwent emergency ventricular drainage, but died of cerebral herniation. The second patient was a 25-year-old woman who presented with impaired consciousness secondary to DKA. Head computed tomography showed subarachnoid hemorrhage and cerebral edema. No related intraoperative findings were observed; it was concluded that the first computed tomography scan revealed pseudo-subarachnoid hemorrhage. Diabetic ketoacidosis-related cerebral edema develops despite treatment according to guidelines and is difficult to predict. Therefore, adult patients should be treated cautiously during DKA management.


Structural principles for computational and de novo design of 4Fe-4S metalloproteins.
Iron-sulfur centers in metalloproteins can access multiple oxidation states over a broad range of potentials, allowing them to participate in a variety of electron transfer reactions and serving as catalysts for high-energy redox processes. The nitrogenase FeMoCO cluster converts di-nitrogen to ammonia in an eight-electron transfer step. The 2(Fe4S4) containing bacterial ferredoxin is an evolutionarily ancient metalloprotein fold and is thought to be a primordial progenitor of extant oxidoreductases. Controlling chemical transformations mediated by iron-sulfur centers such as nitrogen fixation, hydrogen production as well as electron transfer reactions involved in photosynthesis are of tremendous importance for sustainable chemistry and energy production initiatives. As such, there is significant interest in the design of iron-sulfur proteins as minimal models to gain fundamental understanding of complex natural systems and as lead-molecules for industrial and energy applications. Herein, we discuss salient structural characteristics of natural iron-sulfur proteins and how they guide principles for design. Model structures of past designs are analyzed in the context of these principles and potential directions for enhanced designs are presented, and new areas of iron-sulfur protein design are proposed. This article is part of a Special issue entitled Biodesign for Bioenergetics--the design and engineering of electronic transfer cofactors, protein networks, edited by Ronald L. Koder and J.L Ross Anderson.


Does preoperative nephrostomy increase the incidence of wound infection after nephrectomy?
To determine whether patients with nephrostomy after simple nephrectomy more often had postoperative wound complication than did matched patients without nephrostomy. The hospital records of patients who underwent retroperitoneal simple nephrectomy were evaluated, and the following data were retrieved: age, indication for nephrectomy and nephrostomy insertion, medical history, urine culture, antibiotic regimen, time elapsed from nephrostomy insertion to nephrectomy, surgical technique, type of complication, time elapsed from surgery to complication, treatment, and outcome of complications. Thirty-one patients (mean age 57.9 years, +/-SE 3.0) were evaluated. Seven (31.8%) of the 22 patients without nephrostomy (group 1 ) had wound infection compared with 7 (77.7%) of the 9 patients with nephrostomy (group 2) (P <0.05). All 9 group 2 patients had infected urine compared with 11 of the 22 in group 1 (P <0.05). Complications were apparent within a median time of 1 month (+/-SD 0.9) from surgery in group 2, whereas the median time to complication was 4.5 months (+/-SD 3.7, P <0.05) in group 1. Two patients in group 2 died of wound infection and sepsis. Both groups were similarly matched for age, indication for nephrostomy and nephrectomy, perioperative and operative techniques, and histologic findings of the removed kidneys. All patients received antibiotic agents at the time of surgery. Patients with nephrostomy inserted because of pyonephrosis or to relieve obstruction who underwent simple nephrectomy because of unrecoverable renal damage had earlier and more frequent wound infections than patients who underwent the identical procedure without nephrostomy. UROLOGY


Influence of IgG subclasses on automated anti-D (Rho) quantification.
Anti-D quantification by both an automated Polybrene method and an automated trypsin-albumin-dextran (TAD) method gave discrepant results in certain cases. These discrepancies, expressed as the polybrene TAD ratio of reactivity (PTR), were related to the IgG subclass of anti-D. Anti-D of the IgG3 subclass showed a higher PTR than IgG1 (0.94 vs 1.65). No difference was shown between G1m(1) and G1m(3) (0.93 and 0.95, respectively) or between G3m(11) and G3m(21) (1.40 and 1.81, respectively) allotypes. The simultaneous use of our automated Polybrene and TAD methods provides information about the anti-D subclass composition.


Assessment of placental perfusion using contrast-enhanced ultrasound: A longitudinal study in pregnant rabbit.
The main aim of the study was to quantitatively characterize placental perfusion using contrast-enhanced ultrasound in a longitudinal study performed in pregnant rabbits. As a secondary objective, we qualitatively assessed the placental perfusion. Three pregnant rabbits were used. Contrast-enhanced ultrasound was performed twice for each dam, either on embryo development (ED) day 14, 21 or 28 of pregnancy, under general anesthesia. Both qualitative and quantitative analyses were performed based on ultrasound assessment. Altogether, data from 29 fetuses were analyzed: 10 at ED-14; 12 at ED-21, and 7 at ED-28. Placental vascularization was mainly peripheral at the earliest gestational age and became gradually homogeneous within the whole placenta as pregnancy progressed. Intensity parameters and relative blood flows significantly increased with gestational age. The wash-in and wash-out rates were correlated and a significant increase in wash-in compared to in wash-out rates was observed with increased gestational age. Contrast-enhanced ultrasound is feasible in animal models and allows qualitative and quantitative analysis of placental vascularization during pregnancy.


Lifetimes and NADH quenching of tryptophan fluorescence in pig heart cytoplasmic malate dehydrogenase.
The time-resolved and steady state fluorescence properties were measured for pig heart cytoplasmic malate dehydrogenase at pH 6.0 and 8.0. The fluorescence decay can be described by two rate processes, according to the functions: I(t) = 0.7e(-t/1.0) + 0.3e(-t/4.4) for the free enzyme and I(t) = 0.7e(-t/0.8) + 0.3e(-t/2.0 for the enzyme . NADH complex. Quenching by NADH of the tryptophan fluorescence is linear. The only effect of pH is to change the association constant for NADH binding; the fluorescence of the free enzyme and the fluorescence quenching by NADH, I-, and acrylamide are unaffected by pH. Thus there are no changes in conformation of the free enzyme or of the NADH complex over the range of pH 6 to 8.


Effect of acute administration of bromocriptine on isoproterenol- and angiotensin II-induced water intake in estrogen-treated rats.
Chronic administration of an estrogenic agent is well known to attenuate the drinking response of rats to treatment with a variety of dipsogenic agents, and to increase plasma concentration of prolactin. Treatment with prolactin is also known to reduce the drinking response to administration of the dipsogenic agent, isoproterenol. Hence, a possibility existed that the antidipsogenic effect of chronic treatment with estrogen was mediated by an increased plasma prolactin concentration. Since bromocriptine, a dopaminergic agonist, is known to reduce plasma prolactin concentration in estrogen-treated rats, it was administered (1.0 mg/kg, IP) 15 min prior to treatment with either isoproterenol (25 micrograms/kg, SC) or angiotensin II (200 micrograms/kg, SC). The results suggest that the antidipsogenic effect of chronic treatment of rats with estradiol benzoate (30.4 and 45.7 micrograms/kg/day) can be reversed, at least partially, by acute administration of bromocriptine.


Phase transition of RNA-protein complexes into ordered hollow condensates.
Liquid-liquid phase separation of multivalent intrinsically disordered protein-RNA complexes is ubiquitous in both natural and biomimetic systems. So far, isotropic liquid droplets are the most commonly observed topology of RNA-protein condensates in experiments and simulations. Here, by systematically studying the phase behavior of RNA-protein complexes across varied mixture compositions, we report a hollow vesicle-like condensate phase of nucleoprotein assemblies that is distinct from RNA-protein droplets. We show that these vesicular condensates are stable at specific mixture compositions and concentration regimes within the phase diagram and are formed through the phase separation of anisotropic protein-RNA complexes. Similar to membranes composed of amphiphilic lipids, these nucleoprotein-RNA vesicular membranes exhibit local ordering, size-dependent permeability, and selective encapsulation capacity without sacrificing their dynamic formation and dissolution in response to physicochemical stimuli. Our findings suggest that protein-RNA complexes can robustly create lipid-free vesicle-like enclosures by phase separation.


Impact of posttransplant cyclophosphamide on the outcome of patients undergoing unrelated single-unit umbilical cord blood transplantation for pediatric acute leukemia.
Umbilical cord blood transplantation (UCBT) from unrelated donors is one of the successful treatments for acute leukemia in childhood. The most frequent side effect of UCBT is peri-engraftment syndrome (PES), which is directly associated with the greater prevalence of acute and chronic graft-versus-host-disease (aGvHD and cGvHD). In haploidentical stem cell transplantation, posttransplant cyclophosphamide (PTCY) has been demonstrated to be an effective method against GvHD. However, the effects of PTCY as a GvHD prophylactic in UCBT had not been investigated. This study aimed to evaluate the effects of PTCY on the outcomes of UCBT for pediatric acute leukemia. This retrospective study included 52 children with acute leukemia who underwent unrelated single-unit UCBT after myeloablative conditioning regimens. The results from the PTCY and non-PTCY groups were compared. The incidence of transplantation-related mortality in non-PTCY and PTCY were 5% and 10% (p = 0.525), respectively. The incidence of relapse in non-PTCY and PTCY were 5% and 23% (p = 0.095), respectively. Second complete remission status (CR2) was an independent risk factor for relapse-free survival (hazard ratio = 9.782, p = 0.001). The odds ratio for sepsis or bacteremia incidence was significantly greater in the PTCY group (9.524, p = 0.017). PTCY group had increased rates of cytomegalovirus activity and fungal infection. The incidence of PES, aGvHD, cGvHD, and hemorrhagic cystitis in the PTCY group was lower than that in the non-PTCY group, although it was not significantly different. Additionally, higher doses of PTCY (29 mg/kg and 40 mg/kg) were associated with lower incidences of aGvHD and severe GvHD (65% and 29%, respectively) than lower doses (93% and 57%, respectively). Engraftment time and graft failure incidence were similar across groups. The results support the safety and efficiency of PTCY as part of PES controlling and GvHD prophylaxis in single-unit UCBT for children with acute leukemia. A PTCY dosage of 29 mg/kg to 40 mg/kg appears to be more effective in GvHD prophylaxis for UCBT patients.


Basal forebrain GABAergic neurons promote arousal and predatory hunting.
Predatory hunting is an important approach for animals to obtain valuable nutrition and energy, which critically depends on heightened arousal. Yet the neural substrates underlying predatory hunting remain largely undefined. Here, we report that basal forebrain (BF) GABAergic neurons play an important role in regulating predatory hunting. Our results showed that BF GABAergic neurons were activated during the prey (cricket)-hunting and food feeding in mice. Optogenetic activation of BF GABAergic neurons evoked immediate predatory-like actions to both artificial and natural preys, significantly reducing the attack latency while increasing the attack probability and the number of killed natural prey (crickets). Similar to the effect of activating the soma of BF GABAergic neurons, photoactivation of their terminals in the ventral tegmental area (VTA) also strongly promotes predatory hunting. Moreover, photoactivation of GABAergic BF - VTA pathway significantly increases the intake of various food in mice. By synchronous recording of electroencephalogram and electromyogram, we showed that photoactivation of GABAergic BF - VTA pathway induces instant arousal and maintains long-term wakefulness. In summary, our results clearly demonstrated that the GABAergic BF is a key neural substrate for predatory hunting, and promotes this behavior through GABAergic BF - VTA pathway.


Design of new inhibitors for cdc2 kinase based on a multiple pseudosubstrate structure.
New inhibitors have been designed for cdc2 kinase based on a multiple pseudosubstrate structure. The new inhibitors have three different structural components: 3,4-bis(indol-3-yl)maleimide, Ac-Cys-(Ser)-Pro-Lys-Lys-NHMe, and ethyloxy group between the two components. Inhibitory activities toward cdc2 and other protein kinases were investigated, and the compound (21) with Ac-Cys-Pro-Lys-Lys-NHMe connected with the triethylene glycol spacer exhibited the most potent inhibition with relatively high selectivity.


["Since his stroke, Théo has become a child like any other"].
Théo is a 7-year-old boy who was diagnosed as having had an ischemic stroke a few hours after birth. Today, despite some sequelae, Théo is doing well. He continues however to be followed medically. His mother describes his care pathway and the support given to the whole family.


Ovariectomy influences the circadian rhythm of locomotor activity and the photic phase shifts in the volcano mouse.
Recently, the relationship between the circadian system and female reproduction has been of great interest; ovarian hormones can modify the amount and distribution of daily activity differently in rodent species. The volcano mouse Neotomodon alstoni is a species in which it is possible to study the circadian rhythm of locomotion, and it offers comparative information about the influence of ovaries on the circadian system. In this study, we used infrared crossings to compare free movement in intact and sham-operated or ovariectomized mice. We analyzed behavioral and endocrine changes related to the estrous cycle and locomotor circadian rhythm in free-running mice and photic phase shifting. Evidence shows that intact mice present a scalloped pattern of daily activity during the estrous cycle. In constant darkness, the ovariectomy reduces the total amount of activity, shortens the free-running circadian period of locomotion and increases photic phase shifts during the early subjective night. During entrainment, the ovariectomized mice increased the amplitude of total activity during the scotophase, and delay the time of activity onset. These results suggest that ovarian hormones in N. alstoni modulate the circadian rhythm of locomotor activity in a species-specific manner.


HIV, Immune Dysfunction, and the Natural History of Anal High-Risk Human Papillomavirus Infection in Gay and Bisexual Men.
Incidence of anal cancer is highest in gay and bisexual men (GBM). Better understanding of the natural history of anal high-risk human papillomavirus (hrHPV) infection is needed for anal cancer prevention. The Study of the Prevention of Anal Cancer was a 3-year study of Australian GBM, aged 35 years or older. We examined incidence, clearance, and risk factors for 13 hrHPV types at baseline and 3 annual visits. In 525 men with ≥ 2 visits, 348 (66.3%) acquired ≥ 1 incident hrHPV infection. HPV16 incidence rates were similar, but non-16 hrHPV incidence was higher in HIV-positive (51.8/100 person years [PY]) than HIV-negative men (36.5/100 PY, P < .001). Annual clearance rates of HPV16 (13.21/100 PY, 95% confidence interval, 10.53-16.56) were lower than for other hrHPV types. hrHPV clearance rates were not associated with HIV overall but were significantly lower in those with a lower nadir CD4 (<200 cells/µL) for HPV16 (P = .015) and other hrHPV types (P = .007). Higher incidence of non-16 hrHPV types, coupled with lower clearance of non-16 hrHPV types in those with past impaired immune function, is consistent with the greater role of non-16 hrHPV in anal cancer in HIV-positive people. ANZCTR365383.


Aerobic degradation of tetrabromobisphenol-A by microbes in river sediment.
This study investigated the aerobic degradation of tetrabromobisphenol-A (TBBPA) and changes in the microbial community in river sediment from southern Taiwan. Aerobic degradation rate constants (k(1)) and half-lives (t(1/2)) for TBBPA (50 μg g(-1)) ranged from 0.053 to 0.077 d(-1) and 9.0 to 13.1 d, respectively. The degradation of TBBPA (50 μg g(-1)) was enhanced by adding yeast extract (5 mg L(-1)), sodium chloride (10 ppt), cellulose (0.96 mg L(-1)), humic acid (0.5 g L(-1)), brij 30 (55 μM), brij 35 (91 μM), rhamnolipid (130 mg L(-1)), or surfactin (43 mg L(-1)), with rhamnolipid yielding a higher TBBPA degradation than the other additives. For different toxic chemicals in the sediment, the results showed the high-to-low order of degradation rates were bisphenol-A (BPA) (50 μg g(-1))>nonylphenol (NP) (50 μg g(-1))>4,4'-dibrominated diphenyl ether (BDE-15) (50 μg g(-1))>TBBPA (50 μg g(-1))>2,2',3,3',4,4',5,5',6,6'-decabromodiphenyl ether (BDE-209) (50 μg g(-1)). The addition of various treatments changed the microbial community in river sediments. The results also showed that Bacillus pumilus and Rhodococcus ruber were the dominant bacteria in the process of TBBPA degradation in the river sediments.


Empiric antibiotic therapy in early periprosthetic joint infection: a retrospective cohort study.
Periprosthetic joint infection (PJI) is a devastating complication following total knee or total hip arthroplasty (TKA/THA). Appropriate empiric antibiotic treatment, initiated directly after debridement and implant retention (DAIR), is suggested to contribute to treatment success. The aim of this study was to describe the microbiology and the antibiotic susceptibility in early PJI to guide future empiric treatment in a region with a low incidence of methicillin-resistant Staphylococcus aureus (MRSA). Consecutive patients who underwent DAIR within 3 months after primary unilateral TKA or THA between January 2011 and December 2018 were retrospectively identified from the hospital electronic health records. Data on causative pathogens, antimicrobial susceptibility and the number of post-operative days until cultures demonstrated bacterial growth were collected. One hundred and eleven early PJIs were identified of which 65 (59%) were monomicrobial and 46 (41%) polymicrobial. Among all isolated pathogens, Staphylococcus aureus (n = 53; 29%) was the most commonly identified pathogen in early PJI without any involvement of MRSA. 72% of PJIs were susceptible to vancomycin which could be increased to around 90% by adding gram-negative coverage. On the 5th postoperative day, bacterial growth was observed in 98% of cases. All gram-negative bacteria demonstrated positive tissue cultures on the 4th postoperative day. Vancomycin combined with ciprofloxacin or a third generation cephalosporin provided the highest antimicrobial coverage of all responsible pathogens identified in early PJI. Empiric treatment of gram-negative treatment can be safely terminated in the absence of gram-negative pathogens after 4 days of culturing in cases without preoperative antibiotic treatment.


Gene expression profiling on acute rejected transplant kidneys with microarray.
To investigate the gene expression profiles in acute allograft rejection of renal transplantation, and identify the markers for the early diagnosis of acute rejection, heterotopic kidney transplantation was performed by using F344 or Lewis donors and Lewis recipients. No immunosuppressant was used. Renal grafts were harvested on days 3, 7, and 14. A commercial microarray was used to measure gene expression levels in day-7 grafts. The expression levels of 48 genes were up-regulated in the allograft in comparison with the isograft control, and interferon-gamma-induced GTPase gene was most significantly up-regulated in allografts. It is concluded that a variety of pathways are involved in organ transplant rejection which is dynamic and non-balanced. IFN-inducible genes, such as IGTP, may play an important role in the rejection. A lot of important factors involved in acute rejection are unnecessary but sufficient conditions for the rejection. We are led to conclude that it is virtually impossible to make an early diagnosis based on a single gene marker, but it could be achieved on the basis of a set of markers.


Bilateral traumatic third nerve palsy.
Bilateral palsy of the third cranial nerve in a head-injured patient is described. Delayed computed tomography scanning demonstrated a midline necrotic lesion within the mesencephalon ventral to the aqueduct. The lesion, possibly the sequela of a focal contusion, involved both third nerve nuclear complexes and caused paralysis of their voluntary as well as autonomic functions. Gaze mechanisms and long tracts appeared to be less heavily damaged. The literature dealing with third nerve palsy, particularly bilateral cases, from traumatic and other origins is briefly reviewed.


Circadian clocks and metabolism.
Circadian clocks maintain periodicity in internal cycles of behavior, physiology, and metabolism, enabling organisms to anticipate the 24-h rotation of the Earth. In mammals, circadian integration of metabolic systems optimizes energy harvesting and utilization across the light/dark cycle. Disruption of clock genes has recently been linked to sleep disorders and to the development of cardiometabolic disease. Conversely, aberrant nutrient signaling affects circadian rhythms of behavior. This chapter reviews the emerging relationship between the molecular clock and metabolic systems and examines evidence that circadian disruption exerts deleterious consequences on human health.


Intrathoracic shunt displacement with massive pleural effusion after successful shunt placement in a hydropic fetus with multilocular macrocystic congenital pulmonary airway malformation.
We successfully performed shunting for a fetus with a multilocular macrocystic lung mass with hydrops at 22 weeks' gestation. Complete resolution of hydrops was achieved; however, at 35 weeks' gestation, the fetus developed acute massive pleural effusion. Fetal ultrasound examination revealed that one end of the shunting tube had migrated downward in the thoracic cavity, which led to fluid draining from the lung cyst. The baby was delivered at term and was discharged following neonatal intensive care management. Intrathoracic displacement of the shunt can occur, followed by massive pleural effusion due to drainage of cystic fluid.


Metabolism of leukotriene C4 in the anesthetized guinea pig.
The distribution and metabolism of [3H] leukotriene (LT)C4 has been studied in the anesthetized guinea pig. The intravenous administration of [3H] LTC4 (1 muCi/kg) to seven guinea pigs showed a rapid vascular clearance of radioactivity with significant metabolism evident at the 15 sec and 1 min time points with material chromatographing like LTC4 (45.6 +/- 7.5%, 35.0 +/- 4.4%). LTD4 (18.4 +/- 5.1%, 33.2 +/- 4.4%) as well as polar material (25.5 +/- 6.0%, 29.7 +/- 4.7%) respectively. The biliary recovery of radioactivity was found to be 74.5 +/- 5.5% n = 4, over 120 min in the guinea pig with less than 1% of radioactivity present in the urine. Examination of the metabolic profile of the biliary radioactivity showed total conversion of LTC4 to LTD4 which was the major metabolite at early time points, and LTD4 as well as LTE4 at later time points. Significant radioactivity which increased with time was also present at the solvent front of the chromatogram indicating the presence of polar biliary metabolites. These results show that the major route of elimination of peptide leukotrienes is through the bile duct in the anesthetized guinea pig and that LTD4 is the major eliminated metabolite in this model.


Ultracentrifugation evidence for a somatostatin-binding protein in serum.
Using a technique of high speed centrifugation of serum and a well validated immunoassay for the measurement of serum somatostatin-like immunoreactivity, we have demonstrated that somatostatin, unlike other peptide hormones, appears to sediment with large molecular weight proteins. When synthetic somatostatin of increasing concentration was incubated with serum prior to ultracentrifugation, a linear plot of concentration of somatostatin added against concentration sedimenting (or apparently bound to protein) revealed an association curve. These data provide further evidence for the existence of a serum-binding protein for somatostatin.


When will tumor gene expression profiling be incorporated into clinical breast cancer decision making?
DNA microarray based gene expression analysis of primary breast cancers is not in general clinical use in spite of the widespread enthusiasm for the approach. However, several studies have identified gene expression patterns, or "signatures" that are predictive of disease outcome and assays based on these findings have become commercially available. Additional studies are required to further define gene signatures that can be used to help tailor the therapy best suited to for an individual patient. Recent gene expression profiling studies using cell line models to identify downstream transcriptional targets of oncogenic signaling pathways may help achieve this goal.


Combined drought and virus infection trigger aspects of respiratory metabolism related to grapevine physiological responses.
In the Mediterranean region, grapevines usually deal with drought during their summer growth season. Concurrently, grapevines are hosts to a large number of viruses from which grapevine leafroll associated virus-3 is one of the most widespread and provokes considerable economic losses in many vineyards. However, information concerning grapevine metabolic responses to the combination of drought and viral infection is scarce. Gas-chromatography coupled to mass-spectrometry based metabolite profiling was used in combination with growth analysis, viral loads and gas exchange data to perform an integrative study of the effects of individual and combined stress in two Majorcan grapevine varieties at two experimental years. Metabolic responses of both varieties to the combination of water stress and virus infection were specific and not predicted from the sum of single stress responses. Correlations between respiration, biomass and key metabolites highlight specific adjustments of respiratory and amino acid metabolism possibly underlying the maintenance of carbon balance and growth in grapevines under stress combination.


Characteristics and outcomes of patients undergoing high-dose chemotherapy and autologous stem cell transplantation admitted to the intensive care unit: a single-center retrospective analysis.
High-dose chemotherapy and autologous stem cell transplantation (ASCT) can be associated with adverse events necessitating treatment on the intensive care unit (ICU). Data focusing on patients admitted to the ICU during hospitalization for high-dose chemotherapy and ASCT are scarce. We thus conducted a single-center retrospective analysis comprising 79 individuals who had high-dose chemotherapy and ASCT between 2014 and 2020 and were admitted to the ICU between the initiation of conditioning therapy and day 30 after ASCT. The median age was 57 years (range: 20-82 years); 38% of patients were female. B-cell non-Hodgkin lymphoma (34%) and plasma cell disorders (28%) were the most common indications for high-dose chemotherapy and ASCT. Sepsis represented the major cause for ICU admission (68%). Twenty-nine percent of patients required mechanical ventilation (MV), 5% had renal replacement therapy, and 44% needed vasopressors. The ICU, hospital, 90-day, and 1-year survival rates were 77.2%, 77.2%, 72.2%, and 60.3%, respectively. Stable disease or disease progression prior to the initiation of high-dose chemotherapy (p = 0.0028) and MV (p < 0.0001) were associated with an impaired survival. A total of 36 patients died during observation. The most frequent causes of death were the underlying malignancy (44%) and sepsis (39%). Taken together, the present analysis indicates a favorable overall outcome for patients admitted to the ICU during hospitalization for high-dose chemotherapy and ASCT. Thus, this patient group should not be denied admission and treatment on the ICU.


A peptidyl transferase ribozyme capable of combinatorial peptide synthesis.
The formation of peptide bonds is a key step in both the chemical and biological synthesis of peptides. The ribozyme can use a wide range of amino acids as its substrate for the dipeptide synthesis. A library containing 29 peptides whose synthesis was catalyzed by this unique ribozyme was analyzed by mass spectrometry. These results implicate that ribozyme may have potential application in the peptide synthesis.


Real-world experience of using combinatorial pharmacogenomic test in children and adolescents with depression and anxiety.
To evaluate the real-world impact of using a commercially available combinatorial pharmacogenomic (CPGx) test on medication management and clinical outcomes in children and adolescents treated at a tertiary care psychiatry practice. A retrospective cohort study using our prospectively maintained database of patients undergoing CPGx testing was performed. Only patients with clinical data at the time of ordering CPGx test (pre-baseline), potential medication change visit (baseline) and 8-weeks follow-up (post-baseline) visit were included. Clinical Global Impression (CGI) scores for each visit were calculated. Appropriate statistical analysis, including one-sample t-test, paired t-test and Chi-square test was performed. Based on the inclusion criteria, 281 (75.9%) of the 370 patients with CPGx testing were included. Their mean age was 15.8 ± 4.5 years (111 females; 39.5%). The average number of medications significantly increased to 2.4 ± 1.2 on the post-baseline visit [t(280) = 8.34, p < 0.001). Medications were added in 123 (43.7%), replaced in 92 (32.7%) patients and remained unchanged in rest. There was no significant association between medication-related adverse effects and psychotropic medication change group (p = 0.27). The study population showed a significant improvement (p < 0.001) in the CGI severity, efficacy, and global improvement indices. In our experience of using CPGx test in a large cohort of children and adolescents during routine clinical practice, three-quarter of them underwent medication change. Additionally, we noted an improvement in clinical outcomes without impacting adverse effects. While the role of clinical judgement in medication changes in our cohort is likely, CPGx may supplement clinical decision making. However, the best use and benefit of CPGx in routine clinical practice needs further investigation.


The radial artery in coronary surgery: a 5-year experience--clinical and angiographic results.
The radial artery (RA) has been used extensively by us as a way of reducing the use of the saphenous vein. It has been hoped that the RA will maintain greater late patency than the saphenous vein. We evaluated our initial 5-year experience with the RA in coronary surgery. We studied 6,646 consecutive patients who had a single RA (4,872), or bilateral RA (1,774), as coronary grafts, from June 1995 to June 2000. Angiograms were performed mostly in symptomatic patients, or as part of a research project in asymptomatic patients. The patients' mean age was 65.1 years; 23% had diabetes, 14% had unstable angina, and 42% had prior myocardial infarction. An average of 3.3 grafts per patient were performed, 87% from arterial conduit. Conduits used were RA (8,420), left internal thoracic artery (6,296), and right internal thoracic artery (1,076). Operative mortality occurred in 58 (0.9%) patients, stroke in 92 (1.4%), deep sternal infection in 97 (1.4%), reoperation for hemorrhage in 56 (0.9%), and myocardial infarction in 52 (0.8%). Peak mean postoperative creatine kinase MB (CKMB) was 16.5 IU/L. Two patients developed fingertip ischemia. Postoperative angiographic RA patency was 90.2% (333 of 369 distal anastomoses). Good early clinical and angiographic results can be achieved by using the RA in coronary surgery.


VEGF signaling governs the initiation of biliary-mediated liver regeneration through the PI3K-mTORC1 axis.
Biliary epithelial cells (BECs) are a potential source to repair the damaged liver when hepatocyte proliferation is compromised. Promotion of BEC-to-hepatocyte transdifferentiation could be beneficial to the clinical therapeutics of patients with end-stage liver diseases. However, mechanisms underlying the initiation of BEC transdifferentiation remain largely unknown. Here, we show that upon extreme hepatocyte injury, vegfaa and vegfr2/kdrl are notably induced in hepatic stellate cells and BECs, respectively. Pharmacological and genetic inactivation of vascular endothelial growth factor (VEGF) signaling would disrupt BEC dedifferentiation and proliferation, thus restraining hepatocyte regeneration. Mechanically, VEGF signaling regulates the activation of the PI3K-mammalian target of rapamycin complex 1 (mTORC1) axis, which is essential for BEC-to-hepatocyte transdifferentiation. In mice, VEGF signaling exerts conserved roles in oval cell activation and BEC-to-hepatocyte differentiation. Taken together, this study shows VEGF signaling as an initiator of biliary-mediated liver regeneration through activating the PI3K-mTORC1 axis. Modulation of VEGF signaling in BECs could be a therapeutic approach for patients with end-stage liver diseases.


5-aminolevulinic acid photodynamic therapy enhance the effect of acitretin on squamous cell carcinoma cells: An in vitro study.
Squamous cell carcinoma (SCC) remains the second most common nonmelanoma skin cancer (NMSC) worldwide. Both acitretin and 5-Aminolevulinic acid mediated photodynamic therapy (ALA-PDT) have validated effect on SCC. However, the effects of both treatmens remain limited, and there has been no report concerning the potential synergistic effect of both treatments for SCC. To investigate the cytotoxic effect of acitretin on SCL-1 cells, and whether ALA-PDT enhances this effect. CCK-8 and trypan blue exclusion array were used to detect the cell cytotoxicity after acitretin treatment with different concentrations (1.6 × 10-4mg/mL, 1.6 × 10-3 mg/mL, 1.6 × 10-2mg/mL and 1.6 × 10-1mg/mL) for 24 h, 48 h and 72 h. Flow cytometry and trypan blue exclusion assay were used to detect the apoptosis and viability of SCL-1 cells after treated with acitretin, ALA-PDT and ALA-PDT immediately followed by acitretin. Independent sample t test was used to analyze the different incubation time of acitretin and acitretin combined with ALA-PDT on SCL-1 cells. Bonferroni Test One-way Anova method was used to analyze the effect of different treatment on the SCL-1 cells. A significant cytotoxic effect was observed after acitretin treatment, in an acitretin concentration-dependent manner within the range of 1.6 × 10-4mg/mL to 1.6 × 10-1mg/mL and an acitretin incubation time-dependent manner within 24 h-72 h. The total apoptosis rate and dead cells rate in group of ALA-PDT combined with acitretin were both significantly higher than that of acitretin, ALA-PDT group. A stronger apoptotic and cytotoxic effect detected 24 h after treated with acitretin than that of 12 h was observed in this study. Acitretin has a cytotoxic effect on SCL-1 cells, and ALA-PDT treatment enhances the the cytotoxic effect of acitretin.


Short-term and long-term effects of small incision lenticule extraction (SMILE) on corneal endothelial cells.
Small incision lenticule extraction (SMILE) is being used increasingly as a novel approach to correct refractive errors with femtosecond laser. The study was aimed to investigate the corneal endothelial changes 1 day (short term) and 1 year (long term) after SMILE procedure. In this prospective study, a total of 56 eyes of 30 patients with myopia ranging from -3.25 to -8.25 diopters (D) and cylinder up to -3.50D were treated by SMILE. Postoperative uncorrected distance visual acuity (UDVA) and complications were assessed. Endothelial cell density (ECD), the coefficient of variation (CV), and the percentage of hexagonal cells were measured using a noncontact specular microscope preoperatively and at 1 day and 1 year postoperatively. The estimated residual stromal thickness (RST) of each patient was recorded. The median UDVA improved significantly from 1.00logMAR preoperatively to -0.10logMAR both at 1 day and 1 year (p<0.001). No eyes developed corneal edema or other complications during the follow-up period. There were no significant changes in the ECD, CV or the percentage of hexagonal cells at any visit point (all p>0.05). The mean estimated RST was 355.1±32.2μm (range 290-429μm). Endothelial cell changes in terms of ECD, CV or the percentage of hexagonal cells were not correlated with the estimated RST (p>0.05). SMILE yielded improvement in visual acuity and no adverse effects to corneal endothelial cells were found. However, further studies with a lower preoperative ECD and deeper lenticule extraction are needed to conduct.


An analysis of the blood pressure and safety outcomes to renal denervation in African Americans and Non-African Americans in the SYMPLICITY HTN-3 trial.
SYMPLICITY HTN-3, the first trial of renal denervation (RDN) versus sham, enrolled 26% African Americans, a prospectively stratified cohort. Although the 6-month systolic blood pressure (SBP) reduction in African Americans (AAs) was similar in the RDN group (-15.5 ± 25.4 mm Hg, n = 85 vs. -17.8 ± 29.2, n = 49, P = .641), the sham SBP response was 9.2 mm Hg greater (P = .057) in AAs than non-AAs. In multivariate analyses, sham SBP response was predicted by an interaction between AA and a complex antihypertensive regimen (at least one antihypertensive medication prescribed ≥3 times daily), while in the RDN group, SBP response was predicted by an interaction between AA race and baseline BP ≥ 180 mm Hg. AA race did not independently predict SBP response in either sham or RDN. There appears to be effect modification by race with individual-level patient characteristics in both treatment arms that affect the observed pattern of SBP responses.


Memories following physical trauma in patients treated in the ICU: does gender and head injury make a difference?
The aim of this study was to explore differences between male and female survivors and describe the relation between head injury and memories in men and women. This multi-centre study included 239 trauma patients who answered a self-administered questionnaire (ICUM tool) 6-18 months after discharge from the ICU. Few gender differences were found regarding their memories before, during and after the ICU stay. The univariate and multivariate analyses showed that more women than men reported memories from the time in hospital before being admitted to the ICU. Women also recalled a significantly higher number of memories of feelings than men. Furthermore, patients without head injury had more memories from the time before and during their stay in the ICU than patients with head injury but did not report more memories from the time after the ICU. The ISS mean score was higher for men than for women, more men than women had renal failure and more men than women received opioid treatment. The conclusion is that gender is of less importance concerning memories from the time before, during and after the stay in the ICU.


Aortic origin of right pulmonary artery: successful surgical correction in three consecutive patients.
Over a period of four years (1981 through 1984), 3 infants with aortic origin of the right pulmonary artery were seen in our surgical unit. Two infants had a persistent ductus arteriosus, which arose from the left subclavian artery in 1 of them who also had a right-sided aortic arch. There were no other associated abnormalities. Preoperative diagnosis was established by echocardiography in 2 infants. Anatomical surgical correction was undertaken in all 3 infants under cardiopulmonary bypass. There were no operative deaths. One child required reoperation at 15 months for anastomotic stenosis. All 3 children were well two to five years postoperatively with scintillographic evidence of normal ventilation and perfusion. Our experience indicates that this rare but severe congenital cardiac anomaly is easily diagnosed by echocardiography, and confirms that it is eminently amenable to surgical correction.


From neurotransmitter to gene: identifying the missing links.
Survival demands adaptation to changes in the environment, and some of the most important adaptations are accomplished by the nervous system. Adaptive processes such as learning and memory are mediated by complex intracellular adjustments that result, at least in part, from changes in the levels of certain proteins. The involvement of neurotransmitters in the regulation of protein biosynthesis has recently become the focus of much investigation. Questions that are beginning to be explored include: how do neurotransmitters regulate the expression of specific genes in various cell populations? How can a gene product be regulated in opposing or coordinated fashion by different stimuli? How does a neurotransmitter exert differential regulatory influences in the same cell type under different growth conditions? Teresa Esterle and Elaine Sanders-Bush examine these questions with an emphasis on the studies that explore the ability of neurotransmitters to have diverse actions on the same target gene in different tissues or on different genes in the same cell.


Outcomes and Readmissions after Left Atrial Appendage Occlusion in Octogenarians: A Contemporary Analysis.
The outcomes of left atrial appendage occlusion (LAAO) with the Watchman device in octogenarians are unknown as this population was underrepresented in major clinical trials. This study aims at examining the causes and outcomes of readmission after LAAO. A retrospective cohort study based on the National Readmission Database in the United States. Patients aged ≥80 years, admitted between January 2016 and December 2018, with the primary diagnosis of atrial fibrillation or flutter or who had LAAO were included in the study. Patients who died during index admission were excluded. We used the National Readmission Database and International Classification of Diseases, Tenth Revision, Clinical Modification (ICD-10-CM) codes to identify study participants. Data analysis was performed through Stata, version 17. Univariate and multivariate aggression analysis was used to adjust for confounders using Student t tests and χ2 tests. We identified 491,329 patients on anticoagulation (AC) and 2030 patients who underwent LAA closure. Neither group differed regarding hypertension, previous myocardial infarction, or valvular heart disease. All-cause readmissions were lower in the LAAO group at 45 days (adjusted P < .01). All-cause readmissions at 45 and 90 days were similar in both groups. There was an increase in gastrointestinal bleeding (GIB) readmissions in the LAAO at 45 (P < .01), 90 (P < .01), and 180 (P < .01) days. There was no difference in GIB readmission between the 2 groups. There was no also difference in stroke or intracranial hemorrhage rates between the 2 groups throughout the follow-up period. In octogenarians who received LAAO, the rate of GIB increased during the first 6 months after the procedure; however, it was not different from that of AC after that. Special attention should be given to the antithrombotic regimens after LAAO to avoid bleeding in this vulnerable patient population.


Regional localization of the gene coding for the GM2 activator protein (GM2A) to chromosome 5q32-33 and confirmation of the assignment of GM2AP to chromosome 3.
The gene coding for the GM2 activator protein (GM2A) was previously mapped by us to chromosome 5 by an ELISA-based technique. Here we confirm this assignment using a PCR analysis of somatic cell hybrids and describe a regional localization to chromosome 5q32-33 by in situ hybridization. We also confirm the assignment of a pseudogene GM2AP to chromosome 3.


Gold(III)-dithiocarbamato complexes induce cancer cell death triggered by thioredoxin redox system inhibition and activation of ERK pathway.
Although gold compounds are now recognized as promising anticancer agents, so far only gold(I) derivatives have been investigated for this purpose, whereas the use of gold(III) complexes has been hampered by their poor stability under physiological conditions. We have therefore carried out studies on selected gold(III) anticancer agents, showing enhanced stability due to the presence of chelating dithiocarbamato ligands. We found that they induce cancer cell death through both apoptotic and nonapoptotic mechanisms. They also inhibit thioredoxin reductase activity, generate free radicals, modify some mitochondrial functions, and increase ERK1/2 phosphorylation. Based on our results, we propose and discuss a working model suggesting that deregulation of the thioredoxin reductase/thioredoxin redox system is a major mechanism involved in the anticancer activity of the investigated gold(III)-dithiocarbamato complexes.


The longitudinal relationship between processing speed and cognitive ability: genetic and environmental influences.
Goals of the present study were to investigate the relationship between age changes in speed and cognition and the genetic and environmental influences on that relationship. Latent growth models and quantitative genetic methods were applied to data from the Swedish Adoption/Twin Study of Aging. The sample included 778 individuals from both complete and incomplete twin pairs who participated in at least 1 of 4 testing occasions over a 13-year-period. Four factors were constructed from 11 cognitive measures: verbal, spatial, memory, and processing speed. Results indicate that for measures of fluid abilities, the explanatory value of processing speed is paramount for both mean cognitive performance and acceleration with age. A significant proportion of the genetic influences on cognitive ability arose from genetic factors affecting processing speed. For measures of fluid abilities, it is not the linear age changes but the accelerating age changes in cognition that share genetic variance with processing speed.


Sinusoidal intrathecal infusion for assessment of CSF dynamics in kaolin-induced hydrocephalus.
To evaluate whether changes of CSF outflow resistance and compliance in hydrocephalus can be assessed by an intrathecal infusion which is performed at a sinusoidal varying rate. Hydrocephalus was produced in 10 Sprague Dawley rats by instillation of 0.0375 g of kaolin in 0.9% saline into the cisterna magna. Measurements were performed 4 weeks later: With each animal both, three successive constant rate infusions (0-0.02 ml/min) and a sinusoidal infusion (0-0.02 ml/min, frequency 0.006 Hz) were performed. 6 normal animals served as control. The pressure recordings of both infusion techniques were used for the assessment of the CSF outflow resistance. The time constant and the pressure volume index were calculated only from the sinusoidal input testing. The sinusoidal test as well as the constant rate infusion both demonstrated a severe impairment of CSF absorption. By the sinusoidal input, a decreased compliance was confirmed additionally. Thus, the sinusoidal infusion test demonstrated a high resistance and low compliance hydrocephalus in the kaolin-treated group. A simple graphical procedure is presented which allows an easy assessment of CSF dynamics by the sinusoidal infusion test.


Regulatory oversight of the iron trade: posttranscriptional regulation in yeast.
Though a required element, iron is toxic in high concentration. New evidence reveals pathways for controlling the abundance of mRNAs encoded by iron utilization and distribution genes. These studies demonstrate sophisticated systems that limit iron uptake under conditions of abundance and lead to metabolic remodeling when iron is limited.


[Oral hairy leukoplakia induced by topical steroids].
Oral hairy leukoplakia (OHL) is an EBV-associated condition of the oral mucosa, which is often painless. It is found predominantly in HIV-positive patients and is considered a clinical indicator of immunosuppression. OHL has rarely been described in HIV-negative patients, being found most often in association with iatrogenic immunosuppression. OHL induced by topical steroids remains extremely rare. An 81-year-old HIV-negative woman, treated for 3 months with topical steroids for oral lichen planus, developed an asymptomatic white, corrugated, non-removable plaque with vertical folds on the lateral edge of the tongue. Associated oral candidiasis was noted. Based upon histological findings and in situ hybridisation showing numerous EBV-infected epithelial cells, a diagnosis of oral hairy leucoplakia was made. To our knowledge, we report herein only the second recorded case of OHL induced strictly by topical steroids. Self-medication and poor adherence to dosage recommendations were noted in the patient's medical history. Physicians must be aware of the rare but nevertheless possible adverse events associated with topical steroid use, particularly when such medication is prescribed over a long period for inflammatory diseases of the oral mucosa.


Transcatheter arterial chemoembolization in the treatment of hepatoblastoma.
The prognosis of hepatoblastoma is poor unless the tumor is completely resected. Various types of chemotherapy have been developed to increase its resectability. Recently, transcatheter arterial chemoembolization (TACE) has been developed for the treatment of unresectable adult hepatoma with favorable results. The authors applied this procedure to hepatoblastoma in infants and children. TACE was performed in eight hepatoblastoma cases. After an intraarterial catheter was inserted into the main feeding artery of the tumor, injection of adriamycin or THP-adriamycin (20 to 30 mg/m2) dispersed in lipiodol and cisplatin (40 to 60 mg/m2) followed by embolization using Gelfoam pieces was performed. Effects of TACE were evaluated according to shrinkage of tumor mass on imaging examinations, alpha-fetoprotein (AFP) levels, and pathological findings of the surgical specimens 4 weeks after TACE. A marked reduction in tumor size associated with a decrease in AFP level occurred 1 month after the treatment. Tumor shrinkage ranged from 0.9% to 45.0% with a mean value of 25.8%. AFP levels decreased by 0.2% to 11.9% with a mean level of 4.6% from initial levels. In addition, there was no marked chemotherapeutic agent-induced toxicity noted during the observation period. Resection of the tumors was performed safely after TACE in all cases. Pathological examination showed massive necrosis in the surgical specimens, and the mean percentage of necrotic area within the tumor was 71.1%. Two patients died of extensive lung metastasis 2 months and 3 years after the operation, respectively. The remaining six were doing well and free of disease at a mean follow-up period of 50 months. TACE is an effective, safe, and useful method for the initial treatment of hepatoblastoma.


Cardiomyocyte-Targeting Peptide to Deliver Amiodarone.
Amiodarone is underutilized due to significant off-target toxicities. We hypothesized that targeted delivery to the heart would lead to the lowering of the dose by utilizing a cardiomyocyte-targeting peptide (CTP), a cell-penetrating peptide identified by our prior phage display work. CTP was synthesized thiolated at the N-terminus, conjugated to amiodarone via Schiff base chemistry, HPLC purified, and confirmed with MALDI/TOF. The stability of the conjugate was assessed using serial HPLCs. Guinea pigs (GP) were injected intraperitoneally daily with vehicle (7 days), amiodarone (7 days; 80 mg/kg), CTP-amiodarone (5 days; 26.3 mg/kg), or CTP (5 days; 17.8 mg/kg), after which the GPs were euthanized, and the hearts were excised and perfused on a Langendorff apparatus with Tyrode's solution and blebbistatin (5 µM) to minimize the contractions. Voltage (RH237) and Ca2+-indicator dye (Rhod-2/AM) were injected, and fluorescence from the epicardium split and was captured by two cameras at 570-595 nm for the cytosolic Ca2+ and 610-750 nm wavelengths for the voltage. Subsequently, the hearts were paced at 250 ms with programmed stimulation to measure the changes in the conduction velocities (CV), action potential duration (APD), and Ca2+ transient durations at 90% recovery (CaTD90). mRNA was extracted from all hearts, and RNA sequencing was performed with results compared to the control hearts. The CTP-amiodarone remained stable for up to 21 days at 37 °C. At ~1/15th of the dose of amiodarone, the CTP-amiodarone decreased the CV in hearts significantly compared to the control GPs (0.92 ± 0.05 vs. 1.00 ± 0.03 ms, p = 0.0007), equivalent to amiodarone alone (0.87 ± 0.08 ms, p = 0.0003). Amiodarone increased the APD (192 ± 5 ms vs. 175 ± 8 ms for vehicle, p = 0.0025), while CTP-amiodarone decreased it significantly (157 ± 16 ms, p = 0.0136), similar to CTP alone (155 ± 13 ms, p = 0.0039). Both amiodarone and CTP-amiodarone significantly decreased the calcium transients compared to the controls. CTP-amiodarone and CTP decreased the CaTD90 to an extent greater than amiodarone alone (p < 0.001). RNA-seq showed that CTP alone increased the expression of DHPR and SERCA2a, while it decreased the expression of the proinflammatory genes, NF-kappa B, TNF-α, IL-1β, and IL-6. Our data suggest that CTP can deliver amiodarone to cardiomyocytes at ~1/15th the total molar dose of the amiodarone needed to produce a comparable slowing of CVs. The ability of CTP to decrease the AP durations and CaTD90 may be related to its increase in the expression of Ca-handling genes, which merits further study.


Efficacy of inverted internal limiting membrane flap technique for the treatment of macular hole in high myopia.
To determine the effect of pars plana vitrectomy (PPV) with inverted internal limiting membrane (ILM) flap technique for macular hole with or without retinal detachment in highly myopic eyes. Retrospective, interventional case series. Ten eyes of 10 patients with macular hole with (4 eyes) or without (6 eyes) retinal detachment in high myopia (axial length more than 26.5 mm) were treated by PPV with inverted ILM flap technique. Macular hole closure was observed in 8 eyes (80%) following the initial surgery (in 5 eyes without retinal detachment and in 3 eyes with retinal detachment). In 4 eyes with retinal detachment caused by macular hole, retinas of 3 eyes were reattached by the initial surgery. Postoperative best-corrected visual acuity improved by more than 2 lines in 5 eyes (50%), was unchanged in 4 eyes (40%), and worsened by more than 2 lines in 1 eye (10%). Inverted ILM flap technique might contribute to a high closure rate of macular hole and be a preferable adjuvant to the treatment of macular hole in high myopia with or without retinal detachment.


A novel and efficient synthesis of diverse dihydronaphtho[1,2-b]furans using the ceric ammonium nitrate-catalyzed formal [3 + 2] cycloaddition of 1,4-naphthoquinones to olefins and its application to furomollugin.
A novel approach was developed for the synthesis of diverse dihydronaphtho[1,2-b]furans from 1,4-naphthoquinones and olefins in the presence of ceric ammonium nitrate. This reaction provides a rapid route for the synthesis of a variety of dihydronaphtho[1,2-b]furans and naphtho[1,2-b]furans bearing different substituents. This methodology was also used to synthesize the biologically important natural product furomollugin in only 2 steps.


Long-term outcome following imatinib therapy for chronic myelogenous leukemia, with assessment of dosage and blood levels: the JALSG CML202 study.
A prospective multicenter Phase II study was performed to examine the efficacy and safety of imatinib therapy in newly diagnosed Japanese patients with chronic-phase CML. Patients were scheduled to receive imatinib 400 mg daily. Plasma imatinib concentrations were measured by liquid chromatography-tandem mass spectrometry. In 481 evaluable patients, estimated 7-year overall survival (OS) and event-free survival (EFS) at a median follow-up of 65 months were 93% and 87%, respectively. Because imatinib dosage was reduced in many patients due mainly to adverse events, subgroup analysis was performed according to the mean daily dose during the first 24 months of treatment: ≥360 mg (400-mg group; n = 294), 270-359 mg (300-mg group; n = 90) and <270 mg (200-mg group; n = 67). There were no significant differences in OS and EFS between the 300- and 400-mg groups; however, cumulative rates of complete cytogenetic and major molecular responses differed significantly between the two groups. There were no significant differences in mean imatinib trough levels between these two groups for the patients in whom trough levels had been measured. Survival and efficacy in the 200-mg group were markedly inferior to the former two groups. These results suggest that, although a daily dose of 400 mg imatinib is associated with better outcomes, 300 mg imatinib may be adequate for a considerable number of Japanese patients who are intolerant to 400 mg imatinib. Blood level monitoring would be useful to determine the optimal dose of imatinib.


Point-of-Care Chest Ultrasonography as a Diagnostic Resource for COVID-19 Outbreak in Nursing Homes.
Bedside chest ultrasonography, when integrated with clinical data, is an accurate tool for improving the diagnostic process of many respiratory diseases. This study aims to evaluate the feasibility of a chest ultrasonographic screening program in nursing homes for detecting coronavirus disease-19 (COVID-19)-related pneumonia and improving the appropriateness of hospital referral of residents. Pragmatic, descriptive, feasibility study from April 2 to April 9, 2020. A total of 83 older residents (age 85 ± 8) presenting mild to moderate respiratory symptoms and not previously tested for COVID-19, residing in 5 nursing homes in Northern Italy. Chest ultrasonography was performed at the bedside by a team of hospital specialists with certified expertise in thoracic ultrasonography, following a systematic approach exploring 4 different areas for each hemithorax, from the anterior and posterior side. Presence of ultrasonographic signs of interstitial pneumonia, including comet-tail artifacts (B-lines) with focal or diffuse distribution, subpleural consolidations, and pleural line indentation, was detected. The specialist team integrated ultrasonography data with clinical and anamnestic information, and gave personalized therapeutic advice for each patient, including hospital referral when needed. The most frequent reasons for ultrasonographic evaluation were fever (63% of participants) and mild dyspnea (40%). Fifty-six patients (67%) had abnormal ultrasonographic findings. The most common patterns were presence of multiple subpleural consolidations (32 patients) and diffuse B-lines (24 patients), with bilateral involvement. A diagnosis of suspect COVID-19 pneumonia was made in 44 patients, and 6 of them required hospitalization. Twelve patients had ultrasonographic patterns suggesting other respiratory diseases, and 2 patients with normal ultrasonographic findings were diagnosed with COPD exacerbation. In nursing home residents, screening of COVID-19 pneumonia with bedside chest ultrasonography is feasible and may represent a valid diagnostic aid for an early detection of COVID-19 outbreaks and adequate patient management.


Adsorption of concanavalin A on human platelets.
A physiological cell surface adsorption system approach is investigated on human platelets utilizing mathematical modeling. Monodispersed washed platelets are freshly collected in an isotonic buffer as a suspension utilizing a gel filtration technique. Concanavalin A is used as a glycoprotein receptor adsorbate in the adsorption studies. Three mathematical models are proposed based on simple chemical equilibrium reactions between adsorbate and cell surface receptors in an effort to explain concanavalin A - platelet surface glycoprotein interaction. Model I assumes that all receptors are undergoing simultaneous surface reactions with the adsorbate and without correlation. Model II reflects a strong correlation between the receptors, when only one receptor is active and the second receptor(s) is nothing but the combination of first receptor-adsorbate complex. Model III assumes the presence of multiple receptors on the cell surface. Only when a specific fraction of the total number of one receptor have reacted, will the other receptor(s) initiate reaction with the adsorbate. The results suggest the existence of at least three major glycoprotein receptors interacting with the lectin, and having different equilibrium constants as indicated in the adsorption isotherm. Model III seems to support best the experimental data of concanavalin A interaction with platelet surface glycoproteins.


Parametric studies of matched filters to enhance the signal-to-noise ratios of LC-MS-MS peaks.
Chromatographic parameters of reference signals employed in matched filter methods have been studied using numerical experiments to improve the signal-to-noise (S/N) ratios of small liquid chromatography (LC) peaks obtained with electrospray tandem mass spectrometers (MS-MS). These parameters include the width, shape, and S/N ratios of chromatographic peaks used as the reference signal profiles. Our results show the effect of reference peak widths on improving the S/N ratio of chromatographic peaks; the influence of reference peak shapes is negligible. To verify simulation results, various reference signals, including analyte peaks of high concentration standards, internal standard peaks, and artificial Gaussian peaks of different widths, have been employed to enhance signal peaks on real liquid chromatography-tandem mass spectrometry (LC-MS-MS) chromatograms via matched filter methods. Our experimental results demonstrate that the S/N ratio enhancement of chromatographic peaks agree with the simulation predictions. These findings, therefore, suggest that regardless of peak shape, a well-smooth peak with a width close to that of the analyte peak is an adequate reference signal, when matched filter methods are used to improve LC-MS-MS chromatograms. Nevertheless, all methods processed LC-MS-MS peaks in this study do not achieve the ideal improvement ratios estimated with simulation results. We attribute this deficiency to spike-like noise, which have considerable low frequency components riding on LC-MS-MS chromatograms. Matched filtering, which works as a low-pass filter in the frequency domain, cannot effectively eliminate low frequency flicker noise contributed by these spikes. In addition, simple median filtering does not provide adequate improvement despite being able to smooth out most spikes in the chromatograms.


Deficient neurogenic innervation of the myenteric plexus with normal submucous plexus involving the entire small and large bowel.
A newborn presented with a picture of intestinal obstruction. Multiple biopsies of the small and large bowel showed an unusual neurogenic innervation. The myenteric plexus of Auerbach was severely depleted of ganglion cells and nerve fibers, while the submucous plexus of Meissner was normally innervated. An ileostomy failed to function and extended trial with experimental smooth muscle stimulant (Cisapride) was equally ineffective. The patient was finally treated by a myectomy from the duodenum to the descending colon with a sigmoid colostomy. This procedure, coupled with a Nissen fundoplication, stopped the vomiting and allowed normal defecation through the colostomy. The patient is presently taking increasing increments of oral fluids with a concomitant decrease in the volume of parenteral nutrition. The myectomy initiated marked hypertrophy of the muscularis mucosa. Could this muscular hypertrophy account for the improvement in bowel function? Possible etiology will be discussed. We caution that rectal submucosal suction biopsy alone may be misleading if normal ganglion cells and nerve fibers are found, yet the patient's clinical symptoms fail to improve. A full thickness bowel wall biopsy is then recommended.


Selection of Lactobacillus plantarum strains to use as starters in fermented table olives: Oleuropeinase activity and phage sensitivity.
Fermented table olives (Olea europaea L.) are largely diffused in the Mediterranean area. Olives are picked at different stages of maturity and after harvesting, processed to eliminate the characteristic bitterness caused by the presence of the oleuropein glucoside and to become suitable for human consumption. The spontaneous fermentation of table olives mainly depends on lactic acid bacteria (LAB), and in particular on Lactobacillus plantarum which plays an important role in the degradation of oleuropein. The hydrolysis of oleuropein is attributed to the β-glucosidase and esterase activities of the indigenous LAB microflora. This study investigated the potential of L. plantarum strains isolated from dairy products and olives to be used as starters for fermented table olives. Forty-nine strains were typed by RAPD-PCR and investigated for the presence of the β-glucosidase (bglH) gene. The full sequence of the bglH gene was carried out. All the 49 L. plantarum strains were also tested for phage resistance. A total of six strains were selected on the basis of genotypic polymorphism, bglH gene sequence analysis, and phage resistance profile. These strains were further characterized to assess the acidifying capability, the growth at different temperatures, the tolerance to different NaCl concentrations, and the oleuropeinolytic activity. Although further characterizations are required, especially concerning the influence on sensory properties, L. plantarum proved to have the potential to be used as a debittering and fermentative agent in starter culture for fermented table olives.


Elevated levels of FN1 and CCL2 in bronchoalveolar lavage fluid from sarcoidosis patients.
Sarcoidosis is a granulomatous systemic inflammatory disease in which more than 90 % of all patients develop pulmonary manifestations. Several gene associations have previously been described, but established and clinically useful biomarkers are still absent. This study aimed to find proteins in bronchoalveolar lavage (BAL) fluid that can be associated with the disease. We developed and performed profiling of 94 selected proteins in BAL fluid and serum samples obtained from newly diagnosed and non-treated patients with sarcoidosis. Using multiplexed immunoassays, a total of 317 BAL and 217 serum samples were analyzed, including asthmatic patients and healthy individuals as controls. Our analyses revealed increased levels of eight proteins in sarcoidosis patients compared to controls. Out of these, fibronectin (FN1) and C-C motif chemokine 2 (CCL2) revealed the strongest associations. In addition, cadherin 5 (CDH5) was found to correlate positively with lymphocyte cell numbers in BAL fluid. Applying a high throughput proteomics screening technique, we found proteins of potential clinical relevance in the context of sarcoidosis.


The impact of selective logging on forest structure and tenrec populations in western Madagascar.
This paper reports on possible effects of selective logging on vegetation structure and its consequences for two tenrec species in a dry forest in western Madagascar. On a small scale semi-mechanized logging of less than 10 m3 per ha alters forest structure significantly. However on a large scale most of these changes are smaller than the variation of forest structure due to natural causes. The most profound ecological effect of logging is a reduction in the number of woody species in the regenerating cohort of trees. Potential longterm consequences of this feature are discussed. Microhabitat variables influencing the habitat utilization of two tenrec species,Echinops telfairi andTenrec ecaudatus, were identified in an area of unlogged forest. The conclusions derived from this analysis were then tested in other parts of the forest which had been modified by selective logging. Logging of the extent described above slightly impairs habitats forE. telfairi but has no effect onT. ecaudatus. Though the latter species is hunted extensively in the logged areas, loggingper se does not threaten the survival of either species.


miR-137-LAPTM4B regulates cytoskeleton organization and cancer metastasis via the RhoA-LIMK-Cofilin pathway in osteosarcoma.
Osteosarcoma (OS) is a rare malignant bone tumor but is one leading cause of cancer mortality in childhood and adolescence. Cancer metastasis accounts for the primary reason for treatment failure in OS patients. The dynamic organization of the cytoskeleton is fundamental for cell motility, migration, and cancer metastasis. Lysosome Associated Protein Transmembrane 4B (LAPTM4B) is an oncogene participating in various biological progress central to cancer biogenesis. However, the potential roles of LAPTM4B in OS and the related mechanisms remain unknown. Here, we established the elevated LAPTM4B expression in OS, and it is essential in regulating stress fiber organization through RhoA-LIMK-cofilin signaling pathway. In terms of mechanism, our data revealed that LAPTM4B promotes RhoA protein stability by suppressing the ubiquitin-mediated proteasome degradation pathway. Moreover, our data show that miR-137, rather than gene copy number and methylation status, contributes to the upregulation of LAPTM4B in OS. We report that miR-137 is capable of regulating stress fiber arrangement, OS cell migration, and metastasis via targeting LAPTM4B. Combining results from cells, patients' tissue samples, the animal model, and cancer databases, this study further suggests that the miR-137-LAPTM4B axis represents a clinically relevant pathway in OS progression and a viable target for novel therapeutics.


Alien balsams, strawberries and their pollinators in a warmer world.
Strawberries are a common crop whose yield success depends on the availability of pollinators. Invasive alien plants, such as Impatiens glandulifera and I. parviflora, are also attractive for bees and hoverflies, respectively, and occur in close proximity to strawberry cultivation areas. The aim of the study was to test whether alien plants may decrease pollination of strawberry cultivation. However, even if the pollinators are abundant, efficiency of their pollination may decrease as a result of revisits of flowers that were already probed. It is addressed by pollinators by scent marking. Moreover, such revisits can be determined by nectar replenishment, which may occur rapidly in nectar-rich flowers. We studied revisits to I. glandulifera by bumblebees and defined the factors that influence the probability of revisits (air temperature; pollinator species; family caste and size; flower area; sun radiation; and time of day). We found that the two alien species decreased the number of pollinators visiting strawberries. Apoidea, Bombini and Syrphidae significantly decreased on Fragaria × ananassa when alien Impatiens were present. We also revealed the influence of increasing air temperature on bumblebee foraging, which was particularly significant for female workers. At very high temperatures (> 37°C), bumblebee males revisited probed flowers less often than female workers. Our results demonstrate that in experimental conditions attractive alien species decrease pollination of strawberries, which may negatively affect production of this crop. Although the results have not been verified in real-life strawberry fields yet, we recommend that alien plant species that share the same pollinators and occur in close proximity of strawberries are controlled. Moreover, we found that revisits of probed flowers may weaken feeding efficiency of bumblebees. If revisits are not induced by nectar replenishment, then global warming may pose a serious threat to the survival of colonies, which may have consequences also for the plants that attract them, e.g., for strawberries.


[Tensometry - development of a measurement technique of the tube function in a temporal bone model].
Middle ear pressure depends widely on the function of the eustachian tube. Tube dysfunction is often a trigger for middle ear diseases like chronic otitis media but also for barotrauma. Patients with impaired tubal function should not be exposed to situations with extreme pressure changes. Until today, there is no valid examining method for long-time measuring of the development of middle ear pressure. The basic idea was to develop a thin flexible film with integrated strain gauges made of platinum and gold, which can be applied directly on the surface of the tympanic membrane. Using these, shifts or distensions of the tympanic membrane in a temporal bone model can be measured and documented. We were able to show that the measuring system was sensitive enough to register pressure variations in the middle ear volume of a temporal bone model. While using a full bridge design of the strain gauge resistors it could be shown that the measuring voltage of the strain gauges were in phase of the pressure measurement curve or according to the positioning in opposite phase. In single resistor mode the measurement were not so positioning depended. Measuring tympanic membrane movement in case of perforation was feasible. To improve the longtime stability of the strain gauge film the next development step will be to make a new design with Konstantan resistors (CU-Ni-Mn). After a testing period, longtime measurements in clinical studies will follow.


Erratum: Cold snare polypectomy of colorectal polyps ≤ 10 mm on clopidogrel: Australian and New Zealand randomized controlled trial.
[This corrects the article DOI: 10.1055/a-1813-1019.].


Fenofibrate impairs liver function and structure more pronounced in old than young rats.
Since old animals are known to accumulate lipids in some organs, we compared effects of fenofibrate (FN) on systemic lipid metabolism, activity of liver marker enzymes and structure in young and old rats. Young and old rats were fed chow supplemented with 0.1 % or 0.5 % FN. After 30 days, intraperitoneal glucose tolerance test (IPGTT) was performed, and blood and liver samples were collected. In young rats, 0.1 % FN, but not 0.5 % FN, decreased serum Chol by 74 %, and did not affect TG levels at either doses. In old rats, 0.5 % FN, but not 0.1 % FN, decreased Chol and TG level by 56 % and 49 %, respectively. In young rats, 0.1 % and 0.5 % FN increased serum activity of ALP by 227 % and 260 %, respectively, and did not affect AST and ALT activities. In old rats, only 0.5 % FN increased serum ALP activity by 150 %, respectively. In old rats, neither dose of FN affected serum AST activity, and only 0.5 % FN increased serum ALT activity by 200 %. The histological examination of liver structure revealed that both doses of FN impaired lobular architecture, expansion of bile canaliculi, and degeneration of parenchymal cells with the presence of cells containing fat droplets; administration of FN increased area occupied by collagen fibers. Although 0.5 % FN decreased serum Chol concentration, it increased serum ALP activity and impaired liver structure in both in both age groups of rats. Thus, FN treatment should be under the control of liver function, especially in older patients.


Green construction of recyclable amino-tannic acid modified magnetic nanoparticles: Application for β-glucosidase immobilization.
The β-glucosidase (BGL) enzyme in food industry is great interest due to its role in food conversion to produce functional food products. In this study, the BGL was covalently immobilized onto amino-tannic acid modified Fe3O4 magnetic nanoparticles (ATA-Fe3O4 MNPs) as biocompatible nanoplatform by modified poly-aldehyde pullulan (PAP) as a cross-linker to enhance the ability and strength of the nanoparticle connection to the enzyme. The properties of support were subsequently characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), Fourier transformed infrared (FTIR), X-ray diffraction (XRD), and vibrating sample magnetometer (VSM). The highest percentage of loading and immobilization yield was obtained with 0.1 mg enzyme/mL citrate buffer (pH 6, 1 M) enzyme solution, carrier solution of 10 mg ATA-Fe3O4/3 mL citrate buffer (pH 6, 1 M), and PAP solution of 20% total reaction system volume. Optimum pH and temperature were found for free (pH 5.0 and temperature 30 °C) and immobilized (pH 6.0 and temperature 40 °C) enzyme. The immobilized BGL maintains its activity to 83% after 10 cycles. Therefore, immobilization of BGL by this method is an efficient procedure to improve the properties of enzyme.


The maternal benefits of corticosteroids with HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome.
To determine the more effective regime in improving hematologic abnormalities associated with HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome, by comparing two different antenatal dosing protocols for dexamethasone (given primarily for fetal lung maturity). A retrospective cohort study of 30 women between 24 and 34 weeks' gestation, diagnosed with HELLP syndrome prior to delivery, who received 2 doses of dexa-methasone 12 mg intramuscularly, given either 24 hours apart or 12 hours apart. There was a shorter time from the first corticosteroid dose to the beginning of improvement with the 12-hour regime for platelet count (11 hours [9, 25 hours] versus 69 hours [41, 112 hours], median [quartiles], P = 0.003) and for alkaline phosphatase (25 hours [5, 44 hours] versus 59 hours [31, 69 hours], median [quartiles], P = 0.02). Worsening recurred from 11 hours, for alanine aminotransferase (ALT) and aspartate transaminase (AST), to 32 hours, for uric acid, after initial improvement. Hematologic improvement occurred in 37% (for ALT) to 67% (for alkaline phosphatase) of women overall. A temporary improvement in the hematologic parameters of HELLP syndrome can occur with antenatal dexa-methasone administration, with the 12-hour regime having quicker onset to improvement in platelet count and alkaline phosphatase.


Neurological manifestations of coronavirus infections, before and after COVID-19: a review of animal studies.
Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This virus, which was first identified in December 2019 in China, has resulted in a yet ongoing viral pandemic. Coronaviridae could potentially cause several disorders in a wide range of hosts such as birds and mammals. Although infections caused by this family of viruses are predominantly limited to the respiratory tract, Betacoronaviruses are potentially able to invade the central nervous system (CNS) as well as many other organs, thereby inducing neurological damage ranging from mild to lethal in both animals and humans. Over the past two decades, three novel CoVs, SARS-CoV-1, MERS-CoV, and SARS-CoV-2, emerging from animal reservoirs have exhibited neurotropic properties causing severe and even fatal neurological diseases. The pathobiology of these neuroinvasive viruses has yet to be fully known. Both clinical features of the previous CoV epidemics (SARS-CoV-1 and MERS-CoV) and lessons from animal models used in studying neurotropic CoVs, especially SARS and MERS, constitute beneficial tools in comprehending the exact mechanisms of virus implantation and in illustrating pathogenesis and virus dissemination pathways in the CNS. Here, we review the animal research which assessed CNS infections with previous more studied neurotropic CoVs to demonstrate how experimental studies with appliable animal models can provide scientists with a roadmap in the CNS impacts of SARS-CoV-2. Indeed, animal studies can finally help us discover the underlying mechanisms of damage to the nervous system in COVID-19 patients and find novel therapeutic agents in order to reduce mortality and morbidity associated with neurological complications of SARS-CoV-2 infection.


[Dynamics of changes in integral parameters of muscle tissue heat resistance after preliminary heating at the damaging temperature in various frog species].
The dynamics of mean heat resistance of the Rana temporaria m. sartorius against heating at 38 degrees C were studied. The following indices were used: heat resistance (HR), coefficient of correlation between the initial HR level of the muscle and the magnitude and direction of its change, and coefficient of correlation between the heat resistance of the paired muscle and HR variability under the influence of preliminary heating at 38 degrees C for 5-180 min. Almost all these indices showed a biphasic pattern of changes with reference to the dose of preliminary treatment. The dynamics of the coefficient of correlation between the initial HR of muscle and its change and the coefficient of correlation between the resistance of paired muscle and HR variability provides much more information about both the pattern of muscle reaction to preliminary heating and muscle functional state than the mean HR level does.


Different evolutionary strategies for the origin of caspase-1 inhibitors.
Caspase 1 (CASP-1) inhibitors share sequence similarity to CASP-1 itself and are all mapped to chr11q22.3. Here we show that these inhibitors are all products of a series of gene duplications that occurred at this locus after the divergence between human and mouse. Surprisingly, stop codons originated independently in all duplicated copies to generate CARD-only proteins with inhibitory activity. We discuss this evolutionary model in the context of both neo- and subfunctionalization.


Low fasting glucose is associated with enhanced thrombin generation and unfavorable fibrin clot properties in type 2 diabetic patients with high cardiovascular risk.
To investigate the effect of low blood glucose on thrombin generation and fibrin clot properties in type 2 diabetes (T2DM). In 165 patients with T2DM and high cardiovascular risk, we measured ex vivo plasma fibrin clot permeation [Ks], turbidity and efficiency of fibrinolysis including clot lysis time [t50%], together with thrombin generation and platelet activation markers in relation to fasting blood glucose. As compared to patients in medium (4.5-6.0 mmol/l, n = 52) and higher (>6.0 mmol/l, n = 75) glucose group, subjects with low glycemia (<4.5 mmol/l, n = 38) had lower Ks by 11% (p < 0.001) and 8% (p = 0.01), respectively, prolonged t50% by 10% (p < 0.001) and 7% (p = 0.016), respectively, and higher peak thrombin generation by 21% and 16%, respectively (p < 0.001 for both). There were no significant differences in Ks and t50% between patients in medium and higher glucose group. In the whole group, a J-shape relationship was observed between glycemia and the following factors: peak thrombin generation, Ks and t50%. Only in patients with HbA1c < 6.0% (42 mmol/mol) (n = 26) fasting glucose positively correlated with Ks (r = 0.53, P = 0.006) and inversely with t50% (r = -0.46, P = 0.02). By multiple regression analysis, after adjustment for age, fibrinogen, HbA1c, insulin treatment and T2DM duration, fasting glycemia was the independent predictor of Ks (F = 6.6, df = 2, P = 0.002), t50% (F = 8.0, df = 2, P < 0.001) and peak thrombin generation (F = 13.5, df = 2, P < 0.0001). In T2DM patients fasting glycemia <4.5 mmol/l is associated with enhanced thrombin formation and formation of denser fibrin clots displaying lower lysability, especially when strict glycemia control was achieved (HbA1c<6.0%).


Extra-large G proteins have extra-large effects on agronomic traits and stress tolerance in maize and rice.
Heterotrimeric G proteins - comprising Gα, Gβ, and Gγ subunits - are ubiquitous elements in eukaryotic cell signaling. Plant genomes contain both canonical Gα subunit genes and a family of plant-specific extra-large G protein genes (XLGs) that encode proteins consisting of a domain with Gα-like features downstream of a long N-terminal domain. In this review we summarize phenotypes modulated by the canonical Gα and XLG proteins of arabidopsis and highlight recent studies in maize and rice that reveal dramatic phenotypic consequences of XLG clustered regularly interspaced short palindromic repeats (CRISPR) mutagenesis in these important crop species. XLGs have both redundant and specific roles in the control of agronomically relevant plant architecture and resistance to both abiotic and biotic stresses. We also point out areas of current controversy, suggest future research directions, and propose a revised, phylogenetically-based nomenclature for XLG protein genes.


Hypoxia induced responses are reflected in the stromal proteome of breast cancer.
Cancers are often associated with hypoxia and metabolic reprogramming, resulting in enhanced tumor progression. Here, we aim to study breast cancer hypoxia responses, focusing on secreted proteins from low-grade (luminal-like) and high-grade (basal-like) cell lines before and after hypoxia. We examine the overlap between proteomics data from secretome analysis and laser microdissected human breast cancer stroma, and we identify a 33-protein stromal-based hypoxia profile (33P) capturing differences between luminal-like and basal-like tumors. The 33P signature is associated with metabolic differences and other adaptations following hypoxia. We observe that mRNA values for 33P predict patient survival independently of molecular subtypes and basic prognostic factors, also among low-grade luminal-like tumors. We find a significant prognostic interaction between 33P and radiation therapy.


Weak link of tendon-bone healing and a control experiment to promote healing.
This study aims to observe the mechanical weak point and histological features of tendon-bone interface after anterior cruciate ligament reconstructive surgery and to explore the tendon-bone healing effects of the platelet-rich gel (PRG) + deproteinized bone (DPB) compound. A total of 48 New Zealand white rabbits were randomly divided into normal group, model (without embedding), experimental (embedded with the PRG + DPB compound), and control (embedded with DPB) groups. The rabbits were executed at 2, 4, 8, and 12 weeks after the operation. Then, micro-computed tomography scan and uniaxial tensile test were conducted. The fractured specimens were subjected to histological observation. At 4, 8, and 12 weeks after the operation, the bone density of the tendon-bone bound section of the experimental group was higher than that of the other groups (P < 0.05). At 4 and 8 weeks, the maximum tensile load of the experimental group was obviously higher than that of the control and model groups (P < 0.05). Histological observation indicated that the tendon-bone interface in the experimental group had more cartilage and bone tissue growing toward the internal tendon, but the fracture layer mainly occurred in the non-ankylosed part. The mechanical weak point of the early tendon-bone interface was in the immature fibrous tissue. The PRG + DPB compound can effectively trigger tendon-bone healing by promoting the maturation and ossification of the tendon-bone tissue. This compound improved the tensile strength of the healing interface and reduced bone tunnel enlargement.


Single-strand conformation polymorphism (SSCP) analysis as a new diagnostic tool to distinguish dorsal-spined larvae of the Elaphostrongylinae (Nematoda: Protostrongylidae) from cervids.
Single-strand conformation polymorphism (SSCP) was used to genetically differentiate morphologically indistinguishable first-stage larvae (L(1)) of the six species of elaphostrongyline nematodes. A partial fragment (317-336bp) of the first internal transcribed spacer (pITS-1) plus 5' flanking region (76bp of the 18S gene) of the nuclear ribosomal DNA (rDNA) was amplified from individual L(1) of known identity and subjected to SSCP. The results showed that the four species of elaphostrongylines found in North American cervids, Parelaphostrongylus tenuis, P. andersoni, P. odocoilei and Elaphostrongylus rangiferi, could be distinguished from one another based on their distinct (i.e. species-specific) SSCP profiles. In addition, E. alces, a species that occurs in moose in Fennoscandinavia, also had a distinct SSCP profile with respect to the other species of elaphostrongylines. However, the SSCP profiles of E. cervi could not be distinguished from those of E. rangiferi because of a lack of interspecific sequence differences in this region of the ITS-1. The distinct SSCP profiles for the other species were consistent with the interspecific differences in ITS-1 sequences, which ranged from 2 (between P. tenuis and P. andersoni) to 59bp (between genera). The pITS-1 SSCP approach was also used to identify unknown elaphostrongyline L(1) from different hosts and localities in North America. The ability to distinguish between L(1) of the four elaphostrongyline species that occur in North American cervids has important diagnostic and epidemiological implications.


HIV-1 Subtype C Vpr Amino Acid Residue 45Y and Specific Conserved Fragments Are Associated with Neurocognitive Impairment and Markers of Viral Load.
There is increasing evidence that HIV-1 viral protein R (Vpr) plays an important role in the pathogenesis of cognitive impairment. We investigated the relationship between HIV-1 subtype C Vpr sequence variation and HIV-associated neurocognitive impairment as measured by global deficit score (GDS) in treatment-naive individuals. We used different bioinformatic tools and statistical models to correlate vpr variation and cognitive function. We identified a tyrosine at position 45 (45Y) as a signature for neurocognitive impairment and histidine (45H) as a signature in the non-impaired individuals. The presence of signature 45Y was associated by 3.66 times higher GDS, 525 times higher plasma viral load, 15.84 times higher proviral load, and 60% lower absolute CD4-T cell count compared with those without the signature. Additionally, we identified four conserved Vpr fragment sequences, PEDQGPQREPYNEWTLE (5-21), LGQYIY (42-47), TYGDTW (49-54), and PEDQGPQREPYNEW (5-18), that were associated with higher plasma viral load and proviral load. The implication of these findings is that variation of Vpr leads to neurocognitive impairment in HIV infection and worsens the progression of disease in general by promoting the production of provirus, promoting HIV replication and depletion of CD4+ T cells in the periphery.