Cystic fibrosis: the delta F508 mutation does not lead to an exceptionally severe phenotype. A cohort study.
In an attempt to ascertain a relationship between genotype and phenotype, we studied the pulmonary and nutritional status of 123 cystic fibrosis patients with known genotype at an age of 8.5-10 years. Patients represent a cohort as they are almost all those born and diagnosed in a given area and period. They were followed at a single centre using uniform diagnostic and treatment protocols. Pulmonary and nutritional status of homozygous delta 508 patients did not differ from that of compound heterozygotes or of patients with other unspecified genotypes. Pulmonary manifestations varied widely in all genotype groups. With the given number of patients, a slightly higher mortality of delta F508 homozygotes could have been coincidental. We conclude that up to the age of 8.5-10 years the severity of pulmonary lesions and nutritional deficiencies is not related to the delta F508 mutation.


Rapid action of cortisol and testosterone on lipogenic enzymes in a fresh water fish Oreochromis mossambicus: short-term in vivo and in vitro study.
Rapid action of steroid hormones on lipid metabolism is not reported so far in any vertebrate. The present study was intended to evaluate the quick actions of cortisol and testosterone on enzymes, namely malic enzyme (ME), glucose-6-phosphate dehydrogenase (G6PDH), and isocitrate dehydrogenase (ICDH) in Oreochromis mossambicus. Cortisol and testosterone produced rapid and opposite effects on the lipogenic enzymes studied. Cortisol significantly decreased the activities of ME, G6PDH, as early as 5 min and ICDH as early as 10 min in vitro (10(-6) M), and 30 min in vivo (0.1 microg/g body wt.) whereas the same doses of testosterone significantly stimulated the activity of all enzymes as early as 5 min in vitro and 30 min in vivo. Actinomycin D treatment did not interfere with the inhibiting effect of cortisol on enzyme activities when measured at 10 min in the in vitro system. The transcriptional inhibitor appeared to partially block the effect of cortisol in vivo. The stimulatory effect of testosterone was insensitive to the action of actinomycin D both in vivo and in vitro. These effects appear to be brought about independently of new protein synthesis because the rapid responses occurred within a latent period of 5-30 min and were insensitive to the action of actinomycin D, suggesting a non-genomic action.


Intravenous gestational cocaine in rats: effects on offspring development and weanling behavior.
Pregnant rats were injected with cocaine (CN; 6 mg/kg) or an equal volume of saline (SAL), via the tail vein, on gestation days 8-20. A third group was untreated (UT). Maternal weight gain was not affected by dam treatment despite slight differences in food intake. Litter characteristics (e.g., litter size, pup weight) did not differ among groups. Indices of fetal mortality were not affected by the treatments. Developmental tests, initiated on postnatal day (PND) 2, indicated slight delays in the negative geotaxic response and eye opening in cocaine-exposed pups. Open-field and tail-flick tests were performed on PND 21. Pups were acutely injected with cocaine (10 mg/kg, IP), saline, or received no treatment before placement in a novel open field; morphine (1.5 mg/kg, SC) or saline was injected prior to the tail flick test. Pups from CN dams exhibited a significant decrease in spontaneous exploratory behavior compared to both controls, and a time-dependent increase in rearing compared to pups from UT dams. The acute cocaine injection prior to placement in the open field did not alter locomotion or rearing among dam treatment groups. However, the acute cocaine injection did increase stereotypy ratings for female pups from CN dams compared to similarly treated males, and females from SAL and UT dams. No differences were observed among groups in the tail-flick test. These data suggest that the IV route of administration provides a viable method of cocaine delivery in pregnant rats, and provides further evidence of the developmental and behavioral teratogenicity of prenatal cocaine exposure.


Distribution and transportation of mercury from glacier to lake in the Qiangyong Glacier Basin, southern Tibetan Plateau, China.
The Tibetan Plateau is home to the largest aggregate of glaciers outside the Polar Regions and is a source of fresh water to 1.4 billion people. Yet little is known about the transportation and cycling of Hg in high-elevation glacier basins on Tibetan Plateau. In this study, surface snow, glacier melting stream water and lake water samples were collected from the Qiangyong Glacier Basin. The spatiotemporal distribution and transportation of Hg from glacier to lake were investigated. Significant diurnal variations of dissolved Hg (DHg) concentrations were observed in the river water, with low concentrations in the morning (8:00am-14:00pm) and high concentrations in the afternoon (16:00pm-20:00pm). The DHg concentrations were exponentially correlated with runoff, which indicated that runoff was the dominant factor affecting DHg concentrations in the river water. Moreover, significant decreases of Hg were observed during transportation from glacier to lake. DHg adsorption onto particulates followed by the sedimentation of particulate-bound Hg (PHg) could be possible as an important Hg removal mechanism during the transportation process. Significant decreases in Hg concentrations were observed downstream of Xiao Qiangyong Lake, which indicated that the high-elevation lake system could significantly affect the distribution and transportation of Hg in the Qiangyong Glacier Basin.


Bis[tris-(pyridin-2-yl)amine]-iron(II) tris-(di-cyano-methyl-idene)methane-diide.
In the title compound, [Fe{(C5H4N)3N}2][C{C(CN)2}3], both ions lie across centres of inversion, with the anion being statistically disordered over two sets of atomic sites having equal occupancy. The cation and anion have approximate and 32 symmetry, respectively, and the Fe-N bond lengths indicate low-spin FeII. A combination of two-centre C-H⋯N and three-centre C-H⋯(N)2 hydrogen bonds link the ions into complex sheets. Several low-occupancy water mol-ecules are present, whose H atoms could not be located: accordingly, the reflection data were subjected to the SQUEEZE procedure [Spek (2015 ▸). Acta Cryst. C71, 9-18].


Nonstructural protein p39 of feline calicivirus suppresses host innate immune response by preventing IRF-3 activation.
Feline calicivirus (FCV) is an important veterinary pathogen that causes acute upper respiratory tract diseases and, occasionally, highly contagious febrile hemorrhagic syndrome in cats. Many viruses have adopted mechanisms for evading IFN-α/β signaling, particularly by directly or indirectly suppressing activation of IRF-3. In this study, we investigated whether nonstructural proteins of FCV possess these mechanisms. When p39, a nonstructural protein of FCV, was transiently expressed in 293T cells, it suppressed IFN-β and ISG15 mRNA production induced by dsRNA. Expression of p39 also suppressed phosphorylation and dimerization of IRF-3 induced by dsRNA. These results suggest that p39 suppresses type 1 IFN production by preventing IRF-3 activation. This may become an important factor in understanding the pathogenesis and virulence of FCV.


Depletion of m6 A reader protein YTHDC1 induces dilated cardiomyopathy by abnormal splicing of Titin.
N6 -methyladenosine (m6 A) is the most prevalent modification in mRNA and engages in multiple biological processes. Previous studies indicated that m6 A methyltransferase METTL3 ('writer') and demethylase FTO ('eraser') play critical roles in heart-related disease. However, in the heart, the function of m6 A 'reader', such as YTH (YT521-B homology) domain-containing proteins remains unclear. Here, we report that the defect in YTHDC1 but not other YTH family members contributes to dilated cardiomyopathy (DCM) in mice. Cardiac-specific conditional Ythdc1 knockout led to obvious left ventricular chamber enlargement and severe systolic dysfunction. YTHDC1 deficiency also resulted in the decrease of cardiomyocyte contractility and disordered sarcomere arrangement. By means of integrating multiple high-throughput sequence technologies, including m6 A-MeRIP, RIP-seq and mRNA-seq, we identified 42 transcripts as potential downstream targets of YTHDC1. Amongst them, we found that Titin mRNA was decorated with m6 A modification and depletion of YTHDC1 resulted in aberrant splicing of Titin. Our study suggests that Ythdc1 plays crucial role in regulating the normal contractile function and the development of DCM. These findings clarify the essential role of m6 A reader in cardiac biofunction and provide a novel potential target for the treatment of DCM.


Immobilization in external rotation combined with abduction reduces the risk of recurrence after primary anterior shoulder dislocation.
We aimed to compare the effectiveness of immobilization in abduction and external rotation vs immobilization in adduction and internal rotation after primary anterior dislocation of the shoulder. The study randomized 102 patients (age range, 15-55 years) with the diagnosis of primary anterior dislocation of the shoulder to receive immobilization in adduction and internal rotation (AdIR, n = 51) using sling and swathe bandage or immobilization in abduction and external rotation (AbER, n = 51) with a stabilizer brace. Patients received a rehabilitation program 3 weeks after the intervention. After a 24-month follow-up, 33.3% in the AdIR group and 3.9% in the AbER group had recurrence (P < .001). The difference in the recurrence rate was greater in the subgroup aged between 31 and 40 years (44.8% in the AdIR group and 3.8% in the AbER group, P < .001). Ten patients in the AbER group (19.6%) and 3 in the AdIR group (5.8%) discontinued shoulder immobilization before 3 weeks (P = .03). In patients without recurrence, the anterior apprehension test was positive in 6 of 34 in the AdIR group (17.6%) and in 4 of 49 in the AbER group (8.1%, P = .19). Immobilization with the shoulder joint in abduction and external rotation is an effective method to reduce the risk of recurrence after primary anterior shoulder dislocations and should be preferred to the traditional method of immobilization in adduction and internal rotation in clinical practice.


Camina por Salud: walking in Mexican-American women.
Forty-six percent of older Mexican-American women report that they do not engage in leisure time physical activity (PA); 38.1% of them are obese. This study (1) evaluated a PA intervention for coronary heart disease risk reduction and (2) determined which variables affect adherence to PA. For 36 weeks, Group I members walked for 3 days a week and Group II members walked for 5 days a week. We measured the participants' total body fat, regional fat, blood lipids, and adherence to PA The walking interventions favorably affected body fat, with significant differences in body mass index reduction, F(2, 16) = 12.86, p = .001. No statistical difference was noted in the anthropometric and blood lipid results between the baseline and 36-week measures.


Neurotensin, a central nervous system peptide: apparent receptor binding in brain membranes.
Neurotensin, a tridecapeptide recently isolated from bovine hypothalamia, has potent pharmacologic effects in several peripheral systems and a regional distribution in rat brain suggestive of a specialized function. 125I-neurotensin binds to membrane preparations from rat brain saturably, reversibly, and with high affinity (apparent KD = 3 nM) under conditions that minimize degradation of the polypeptide. This binding is displaced by neurotensin sequence fragments with relative potencies generally paralleling their potencies in peripheral systems. 125I-neurotensin binding is highest in specific thalamic, cerebral cortical, and hypothalamic areas of rat and calf brain. White matter, brain stem and cerebellar regions have substantially lower amounts of binding. Characteristics of this binding suggest an association with a physiologically revelant neurotensin receptor.


Propofol metabolism is enhanced after repetitive ketamine administration in rats: the role of cytochrome P-450 2B induction.
In a series of ex vivo and in vivo studies we investigated the ability of repetitive ketamine administration to alter the metabolism and anaesthetic effect of propofol and the role of ketamine-mediated P-450 2B induction in rats. Male Wistar rats were pretreated with 80 mg kg(-1) ketamine i.p. twice daily for 4 days. Pentoxyresorufin O-dealkylation (PROD), P-450 2B protein and mRNA were determined. Residual propofol concentration was measured after incubating hepatic microsomes with 100 muM propofol. Sleeping times induced by i.p. 80 mg kg(-1) propofol were determined. Orphenadrine, a P-450 2B inhibitor, was added in both ex vivo and in vivo studies. Finally, serial whole blood propofol concentrations were determined after i.v. infusion of 15 mg kg(-1) propofol. Ketamine pretreatment produced 5.4-, 3.4- and 1.7-fold increases in hepatic PROD activity, P-450 2B protein and mRNA, respectively. Residual propofol concentration was 46% lower after incubation with microsomes from ketamine-pretreated rats than in the control group. The addition of orphenadrine to ketamine-pretreated microsomes produced an increase in residual propofol concentration in a concentration-dependent manner. Ketamine pretreatment reduced propofol sleeping time to 12% of the control, which was reversed by orphenadrine. The whole blood propofol concentration in ketamine-pretreated rats was significantly lower than that of control rats at 1, 2, 4 and 8 min after cessation of propofol infusion. Repetitive ketamine administration enhances propofol metabolism and reduces propofol sleeping time in rats. We suggest that P-450 2B induction may produce ketamine-propofol interaction in anaesthetic practice.


Ketogenic diet decreases circulating concentrations of neuroactive steroids of female rats.
Ketogenic diet (KD) is used to manage intractable epilepsy; however, the mechanisms underlying its therapeutic effects are not known. Steroid hormones, such as progesterone and testosterone, are derived from cholesterol, and are readily 5alpha-reduced to dihydroprogesterone and dihydrotestosterone, which are subsequently converted to 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP) and 3alpha-androstanediol, neuroactive steroids that can influence seizures. The present study examined the effects of the KD on circulating concentrations of these neuroactive steroids, and their precursors, in intact female rats. Thirty-six, 22-day-old female Sprague-Dawley rats (weaned at 21 days) were fasted for 8 hours prior to placement on one of three dietary regimens for 6 weeks: ad libitum chow, calorie-restricted chow, or KD. After 6 weeks of the diet, when six rats in each dietary condition were in diestrus and six were in behavioral estrus, all rats were administered pentylenetetrazole (PTZ, 70 mg/kg, i.p.). The latency and incidence of seizures were recorded by an observer who was uninformed of the estrous cycle and dietary treatment conditions of the rats. Immediately after each test, trunk blood was obtained for later measurement of pregnane (progesterone, dihydroprogesterone, 3alpha,5alpha-THP) and androstane (testosterone, dihydrotestosterone, 3alpha-androstanediol) neuroactive steroid concentrations in plasma by radioimmunoassay. KD tended to lengthen the latency to, and significantly reduced the number of, PTZ-induced barrel roll seizures. KD also significantly reduced plasma levels of the pregnane (dihydroprogesterone, 3alpha,5alpha-THP) and androstane (dihydrotestosterone, 3alpha-androstanediol) 5alpha-reduced metabolites. These data suggest that levels of pregnane and androstane neuroactive steroids, or their precursors, may underlie some of the antiseizure effects of KD.


[High-efficient renaturation of immobilized recombinant C-terminal fragment of human alpha-fetoprotein].
C-terminal fragment of a human oncofetal alpha-fetoprotein (AFP) may be used in targeted cytostatics delivery to malignant cells of many tumors. AFP fragment (from 404 to 595 amino acids residues of a full-sized protein) was cloned and produced in Escherichia coli cells, BL21 strain (DE3) in the form of inclusion bodies. To obtain a functionally active protein, is it necessary to renature the protein. The renaturation procedure of the AFP third domain (rAFP3D) is considerably complicated by the fact that the protein is hydrophobic and contains a large number of S-S bonds. A renaturation technique of rAFP3D immobilized on silicic metal chelate resin has been developed. The yield of renatured C-terminal fragment was no less than 60% with purity on the order of 98%. The developed technique has been applied for the first time for hydrophobic protein with a large number of S-S bonds. The approach can be applied for efficient renaturation of other hydrophobic proteins with a large number of disulfide bonds for scientific and practical purposes.


Association between preoperative biologic use and surgical morbidity in patients with Crohn's disease.
We evaluated the influence of preoperative treatments with biologics on surgical morbidity in patients with Crohn's disease (CD). We reviewed the surveillance data of patients with CD who underwent surgery between April 2018 and April 2021. The possible risk factors for morbidity were analyzed. A total of 305 surgically treated patients were included. Anti-TNF alpha agents and ustekinumab were used in 92 and 27 patients, respectively, within 12 weeks before surgery. There were no cases of mortality. In total, 70/305 (23.0%) patients developed a complication, and 42/305 (13.8%) patients developed a surgical site infection (SSI) (17 incisional SSIs and 35 organ/space SSIs). Current smoking status (OR 3.44), emergent/urgent surgery (OR 6.85), and abdominoperineal resection (APR) (OR 14.93) were identified as risk factors for total complications. Penetrating disease (OR 14.55) was identified as a risk factor for incisional SSIs. Current smoking status (OR 7.09), an American Society of Anesthesiologists (ASA) score greater than 3 (OR 5.85), a postoperative blood sugar level over 155 mg/dL (OR 4.37), and APR (OR 207.95) were identified as risk factors for organ/space SSIs. No correlation between preoperative treatment with biologics and surgical mortality or morbidity was found. However, we should perform further analyses on a larger number of patients because the analyses may be limited by selection bias for treatment and several confounding factors.


Anovaginal and rectovaginal fistulas in Crohn's disease.
Between 1957 and 1985, 886 women with Crohn's disease and an intact distal large bowel were seen at St. Mark's Hospital. Ninety of these patients developed a fistula between the vagina and anus or rectum at an average age of 34 years. The track of the fistula was clearly documented in 80 patients and was extrasphincteric or suprasphincteric in 36, transsphincteric in 42 (high 13, low 29), and superficial in two. Of the 90 patients, 12 (13 percent) were managed throughout without recourse to surgery. Twelve (13 percent) had the fistula laid open or drainage of an abscess as the only surgery. Twelve (13 percent) underwent repair of the fistula and, of these, eight remain symptomatically cured. One has had further symptoms but no surgery while three later underwent proctectomy for rectal disease. In eight patients the colon was removed and the rectum defunctioned and in 34 the rectum was excised as the initial surgery after development of the fistula. The remaining 12 (13 percent) underwent later proctectomy for rectal disease or failed conservative management of the fistula. Extensive colonic involvement, rectal disease, or associated anal lesions were the main reasons for rectal excision in 38 patients. In only ten was the rectovaginal fistula a prominent indication for proctectomy. As medical treatment, repair, or other local surgery were successful in one third of the patients, these options should always be considered in the first instance.


Developmental consequences of emotional abuse and neglect in vulnerable adolescents: A multi-informant, multi-wave study.
Extant research and theory posit that emotional abuse and emotional neglect-exposure is uniquely harmful during adolescence. Yet, these findings are mostly based on mono-informant, retrospective studies with unselected adults that examine emotional maltreatment in the aggregate. This prevents inferences concerning the unique, prospective risk emotional abuse and neglect, as reported by multiple informants, may confer within at-risk, adolescent samples. In response, the present study examined how emotional abuse and emotional neglect-exposure in adolescence uniquely related to psychological symptoms and social impairment. Our aims were tested in a child welfare system (CWS)-involved sample of adolescents (N = 657; AgeM = 12.49 at baseline) who were participating in a longitudinal study. A multi-informant approach was used to assess emotional abuse/neglect and mental health. Physical abuse and lifetime CWS contact represented covariates in growth curve models. Emotional abuse predicted symptoms within informant, such that youth-reported emotional abuse predicted youth-reported internalizing, β = 0.21, p = .001, and externalizing, β=0.35, p = .001, symptoms while parent-reported emotional abuse predicted parent-reported externalizing, β=0.30, p < .001, and internalizing β=0.29, p < .001, symptoms. Meanwhile, youth-reported emotional neglect predicted heightened self-reported internalizing symptoms, β=0.29, p < .001, parent-reported externalizing symptoms, β=0.15, p = .002 and social impairment across youth, β=-0.17, p = .01 and parent, β=-0.24, p < .001, report. This study shows the importance of distinguishing between these maltreatment subtypes in adolescence and provides measurement recommendations for future maltreatment research. The manuscript concludes by discussing adolescent emotional abuse and neglect-exposure as a maintenance, as opposed to causal risk, factors.


Incremental diagnostic value of CMR-derived LA strain and strain rate in dialysis patients with HFpEF.
To test whether left atrial (LA) strain and strain rate add incremental value in the diagnosis of heart failure with preserved ejection fraction (HFpEF) in dialysis patients over clinical and conventional parameters only. HFpEF frequently occurs in dialysis patients, however, the diagnosis of HFpEF is difficult. Although HFpEF is always companied with LA dysfunction, the performance of novel LA parameters, LA strain, and strain rate, in the diagnosis of HFpEF among dialysis patients remains unknown. In the study, 153 dialysis patients (57 without HFpEF and 96 with HFpEF) and 52 healthy controls underwent cardiovascular magnetic imaging (CMR). Three components of LA strain and strain rate, including reservoir, contractile, and booster pump, were assessed via the CMR feature tracking module. Extra diagnostic value was examined by Harrell's C-statistic. Compared with healthy controls and dialysis patients without HFpEF, dialysis patients with HFpEF had significantly impaired LA reservoir (εs) and contractile (εe) strain and strain rate (SRs, SRe), all p < 0.0001. Among these parameters, εs, εe, and SRe showed relatively high accuracy in diagnosing HFpEF among dialysis patients (areas under the curve: 0.84, 0.91, and 0.90, respectively). Reduction of εs, εe, and SRs provided incremental diagnostic value over conventional clinical and echocardiogram parameters. Combined with εs, εe or SRs, the diagnostic performance was further improved (Harrell's C-statistic: 0.83 vs. 0.96, 0.97, and 0.97, respectively, all p < 0.0001). CMR-derived εs, εe, and SRs might add incremental diagnostic value over conventional indexes in diagnosing HFpEF among dialysis patients.


Cleavage of parathyroid hormone in macrophage endosomes illustrates a novel pathway for intracellular processing of proteins.
Most ligands which are taken up by macrophages are transported to lysosomes where they are degraded to their constituents by a concert of acid hydrolases. This process requires a number of intracellular events which result in the transport of ligands from light density endosomes to the more dense lysosomes. In contrast, our studies have shown that macrophages may process some incoming ligands in endosomes (Diment, S., and Stahl, P. D. (1985) J. Biol. Chem. 260, 15311-15317) and that cathepsin D, an aspartyl protease, is localized in these organelles (Diment, S., Leech, M. S., and Stahl, P. D. (1988) J. Biol. Chem. 263, 6901-6907). Using rabbit alveolar macrophages, which can be subjected to subcellular fractionation, we have traced the intracellular transport and processing of bovine parathyroid hormone (PTH-(1-84]. We present evidence that macrophages internalize PTH-(1-84). Once in endosomes the hormone is cleaved to fragments which include a bioactive peptide, PTH-(1-34), and then the fragments are returned to the extracellular medium, without delivery to lysosomes. The entire cycle from initial binding to release of PTH-(1-34) is achieved within 10-15 min, a time period consistent with findings in vivo. Our data provide evidence for a novel route for processing of an endocytosed ligand.


Neglected pulmonary thromboembolism in a traumatic anterior hip dislocation: A case report.
We report the case of treatment delay for an early onset of pulmonary embolism due to thrombosis in the femoral vein occurred in a traumatic anterior hip dislocation. Mistakes interpretations of clinical findings and pre-operative chest CT-angiography has endangered our patient's life because of the medical report was focused only on acute traumatic injuries. After all, we were not able to find in the literature any similar report that could explain the pathogenesis of this early onset and help us in the patient management. We have tried to list a number of practical suggestions to use in emergency room in these situations.


Cellular telephone communication between hospitals and ambulances.
Cellular telephones provide a new means of wireless telephone communication. In an experimental design study comparing the clarity of receiving and transmitting communications with cellular telephones versus conventional emergency medical radio/microwave equipment, cellular telephones proved to be superior in all aspects of clarity and ease. Cellular telephones have numerous cost and equipment advantages over radio equipment and should replace conventional radio/microwave equipment for emergency medical communication in areas where reliable cellular telephone networks are available. This will improve an important facet of prehospital emergency care.


Diagnostic value of B-Type natriuretic peptide and chest radiographic findings in patients with acute dyspnea.
To compare chest radiographic findings and circulating B-type natriuretic peptide (BNP) levels as an adjunct to clinical findings in the diagnosis of heart failure in patients presenting with acute dyspnea. The diagnostic performance of radiographic evidence of cardiomegaly/redistribution and BNP levels > or =100 pg/mL as indicators of heart failure were assessed in 880 patients presenting with acute dyspnea to the emergency departments of five U.S. and two European teaching hospitals. BNP levels were determined by a rapid, point-of-care device. Two blinded cardiologists reviewed all clinical data and categorized patients as to whether they had acute heart failure (n = 447) or not (n = 433). Three-factor analyses showed that BNP levels > or =100 pg/mL contributed significantly to the prediction of heart failure over each of the radiographic indicators. In a multivariate logistic regression analysis, both BNP levels > or =100 pg/mL (odds ratio [OR] = 12.3; 95% confidence interval [CI]: 7.4 to 20.4) and radiographic findings of cardiomegaly (OR = 2.3; 95% CI: 1.4 to 3.7), cephalization (OR = 6.4; 95% CI: 3.3 to 12.5), and interstitial edema (OR = 7.0; 95% CI: 2.9 to 17.0) added significant, predictive information above historical and clinical predictors of heart failure. In patients presenting to the emergency department with acute dyspnea, BNP levels and chest radiographs provide complementary diagnostic information that may be useful in the early evaluation of heart failure.


Molecular and biological features of mollicutes (mycoplasmas).
The small size of the mollicute genome considerably restricts the amount of genetic information available to the organisms. This is reflected in the relatively small number of cell proteins synthesized, the lack of many biosynthetic pathways and the marked dependence on exogenous nutrients for growth. The protein synthesizing machinery of mollicutes resembles that of eubacteria and is sensitive to the same antibiotics, except for rifampicin, to which RNA polymerases of mollicutes appear resistant. The mollicute ribosomes are built of 50 S and 30 S subunits and contain about 50 different proteins and 5 S, 16 S and 23 S rRNA, as in eubacteria. However, the 5 S rRNA in mollicutes appears shorter (107-112 nucleotides) than in eubacteria (116-120 nucleotides). We hybridized restriction endonuclease-digested DNA from a variety of Mycoplasma, Ureaplasma, Acholeplasma and Spiroplasma species with nick-translated probes consisting of defined portions of the rrnB rRNA operon of Escherichia coli and the rRNA operon of M. capricolum. The results suggest the presence of only one or two sets (operons) of rRNA genes in the genome of Mollicutes, a number falling considerably below that of the eubacteria examined so far but resembling that found in archaebacteria. Our data also indicate a marked nucleotide sequence homology along the rrnB rRNA operon of E. coli and the rRNA operons of the various mollicutes, indicating that the rRNA genes in mollicutes are linked in the classical prokaryotic fashion 16 S-23 S-5 S. Each mollicute appeared to possess, on its genome, different flanking sequences adjacent to the rRNA operon(s), resulting in species-specific hybridization patterns.(ABSTRACT TRUNCATED AT 250 WORDS)


Outcome of Liver Transplants Using Donors After Cardiac Death With Normothermic Regional Perfusion.
The incorporation of normothermic regional perfusion (NRP) to donors after cardiac death (DCD) allows the recovery of liver grafts without the deleterious effects on graft survival the super-rapid technique may cause. The aim of the present report is to determine if the use of NRP in Maastricht type III DCD donors achieves short- and medium-term results comparable to donors after brain death (DBD). This is an observational cohort study including 117 liver transplants executed between November 2016 and April 2021, divided into NRP (n = 39) and DBD (n = 78). Donors were younger in the NRP group (NRP 52 vs DBD 59.4 years; P < .005). Liver recipients in each study group were of similar age and severity of liver disease, although the predominant transplant indication in the NRP group was hepatocellular carcinoma. No differences in ischemia times were found. The incidence of early allograft disfunction and primary nonfunction was balanced between NRP and DBD. Eight patients required retransplant, all of them in the DBD group. No differences were found in biliary complications (NRP 12% vs DBD 5%; P = .104). Ischemic cholangiopathy affected a single DBD patient. Graft survival's Kaplan Meier curve shows a better outcome in the NRP group, although the difference did not reach significance (P = .075). The incorporation of perfusion machines, and specifically the NPR in situ, converts suboptimal liver grafts such as DCD into organs comparable to DBDs.


Inhaled isobutyl nitrite compromises T-dependent, but not T-independent, antibody induction.
Habitual abuse of nitrite inhalants has been linked in epidemiological studies with seropositivity to human immunodeficiency virus and, separately, with Kaposi's sarcoma among AIDS patients. Mice exposed to isobutyl nitrite in an inhalation chamber for 45 min/day for 14 days had depressed IgM and IgG antibody responses. The inhibition was dose-dependent at 750-900 ppm, but antibody responses were increased at an intermediate (600 ppm) dose. Gender differences in immunotoxicity were not observed. Antibody responses to a T-independent antigen (DNP-ficoll) were not affected by the immunotoxic exposure, suggesting that B-cells were refractory to the toxic exposure. Toxic exposure to isobutyl nitrite did not selectively deplete a particular spleen cell population, but caused equivalent reductions of T-cells and B-cells. Finally, exposed mice remained immunocompromised for 3-5 days after terminating exposures. Normal immune responses returned by 5-7 days, suggesting that inhibition of cellular function was reversible.


Loss of the cortisol response to naltrexone in Alzheimer's disease.
The administration of a single dose of the opiate antagonist naltrexone (NT) was accompanied by significant elevations in plasma cortisol in normal elderly subjects; in contrast, the cortisol response to NT was absent in individuals of comparable age with Alzheimer's disease (AD). The differential effect of AD on the cortisol response was not accompanied by a significant group difference in plasma prolactin in response to NT administration. Furthermore, this differential cortisol response to NT was not associated with any evident differences in age, sex ratio, plasma levels of naltrexone or its major metabolite beta-naltrexol, or with differences in measures of nonspecific stress, such as plasma free MHPG, pulse, or blood pressure, between the two groups. The absence of the well-characterized cortisol response to NT in AD, together with other reports of abnormal responses to other pharmacological challenges, suggests that neuroendocrine abnormalities might be an important concomitant and possibly a central contributor to the pathophysiology of Alzheimer's disease.


Infrared spectra of metal-free, N',N-dideuterio, and magnesium porphyrins: density functional calculations.
Infrared frequencies and intensities for the metal-free porphyrin (H2Por), N',N-dideuterio porphyrin (D2Por), and magnesium porphyrin complex (MgPor) have been calculated at density functional B3LYP level using the 6-31G(d) basis set. Detailed assignments of the NH and NM vibrational bands in the IR spectra have been made on the basis of comparison between the calculated data and the experimental results. The previous empirical controversial assignments for the bands at 1224, 1110 and 771 cm(-1) for metal-free porphyrin are also clearly interpreted.


Widespread insecticide resistance in head lice to the over-the-counter pediculocides in England, and the emergence of carbaryl resistance.
Increasing resistance to insecticides used for the control of head lice infestation has been documented over the last decade. Treatment failure and tolerance to insecticides have been validated in a number of studies undertaken in several U.K. centres. To establish the extent of insecticide resistance in head lice and acetylcholinesterase activity in the presence of carbaryl in head lice. Head lice were collected from school children in four centres across England (Exmouth, Loughborough, Leeds and South Shields), and tested in their response to the insecticides permethrin, phenothrin, malathion and carbaryl. Data were compared with information collected in Bristol and Bath in 1998 and with susceptible body lice. The activity of louse acetylcholinesterase was measured with and without carbaryl in head lice collected in Bristol, Leeds, Loughborough and in body lice. The efficacy of a 1% carbaryl lotion was compared in children in Bristol and Leeds. Compared with body lice, head lice from all six centres were significantly different in their response (P < 0.0001) to permethrin, phenothrin and malathion after 2-h exposure tests. There were significant differences in louse acetylcholinesterase activity in body lice, and head lice collected in Loughborough and Bristol in the presence or absence of carbaryl (P < 0.001), indicating enzyme inhibition. However, the difference for lice from Leeds was not significant (P = 0.363) suggesting that the enzyme was resistant to carbaryl. Eighty-nine per cent of children treated in Leeds with carbaryl were cured compared with 100% in Bristol. The data suggest head lice resistance is present in many parts of England to over-the-counter products containing synthetic insecticides (permethrin, phenothrin and malathion). They further suggest that resistance is starting to develop to carbaryl in head lice in Leeds and that extensive use of this product would lead to significant resistance.


[Surgical treatment of unstable trochanteric fractures of the femur. I. Evaluation of the surgical treatment of trochanteric fractures of the femur using angular nails].
One hundred and two patients with trochanteric fractures of the femur were treated with angular nail-plates during a four-year period between 1984 and 1987. It is proved that this method is efficient for stable fracture patients, particularly used 24 hours after trauma. Whereas concerning unstable fractures III-th and IV-th degree most results are unsatisfactory. Disadvantage of the method is that patients can not be weight bearing operated leg soon after operation, what has negative influence for healing results.


Ratiometric fluorescent immunochromatography for simultaneously detection of two nitrofuran metabolites in seafoods.
Competitive immunochromatographic assay (ICA) has been widely used in food safety on-site monitoring. However, traditional competitive ICA relies on the change of single-signal intensity to discriminate the results, leading to limited sensitivity and accuracy of naked-eye visual inspection. In this study, we developed a sensitive ratiometric fluorescent immunochromatographic assay (RFICA) judging results based on the color changes of fluorescence. Two different color quantum dot nanobeads (QBs) were introduced as signal labels for rapid and accurate detection of 3-amino-2-oxazolidinone (AOZ) and semicarbazide (SEM) in seafoods. Under the optimal conditions, the visual detection limit for 2-NPAOZ and 2-NPSEM reached up to 1 and 0.5 ng/mL, which were 5- and 14-fold lower than that of traditional single-signal response fluorescent ICA strips. In addition, the RFICA strips were applied to real samples with acceptable recoveries of 81.33 ∼ 99.23 %. These results demonstrate that the developed RFICA platform possesses considerable potential in the on-site detection.


Human health risk assessment of lead (Pb) through the environmental-food pathway.
Drinking water and farm-to-fork pathways have been identified as the predominant environmental pathways associated with human exposure (HE) to Pb. This study integrates a GIS-based survey of metal concentrations in soil and a probabilistic quantitative risk assessment of Pb through the food chain. The case study area was selected in the east of Ireland. A step-wise exposure assessment collated the data for Pb concentration in soil and water media, bioaccumulation of Pb in unprocessed food products, such as potatoes, carrots, green vegetables, and salad vegetables. The daily mean HE to Pb through selected food products was found to be 0.073 mg day-1, where a mean weekly exposure was estimated as 0.0065 mg kg body weight-1 week-1. Multiple risk estimates were used. Hazard Quotient (HQ), Daily Dietary Index (DDI), Daily Intake of Metal (DIM), Health Risk Index (HRI), Target Hazard Quotient (THQ) and Cancer Risk (CR) were found as 0.234 to 0.669, 0.002, 0.0002, 0.020 to 0.057, 0.234 to 0.669, and 0.00001, respectively which signify a low to moderate risk. A sensitivity analysis revealed that intake of potato is the most sensitive parameter of the model, which is positively correlated (coeff. + 0.66) followed by concentration of Pb in the arable soil (+0.49), bioaccumulation in tubers (+0.37), consumption of salad vegetables (+0.20), and consumption of green vegetables (+0.13) (top 5). A back-calculated limit of Pb in the soil (51 mg kg-1) justifies the lower threshold limit of Pb (50-300 mg kg-1) in agricultural soil set by the European Union to mitigate potential bio-transfer into food products. The study concludes there is a low to moderate risk posed by Pb, within the system boundary of the probabilistic model, and highlights the significance of limiting Pb concentrations in the vegetable producing agricultural soil.


The use of monoclonal anti-CEA antibody immunohistochemistry in detecting the origin of oral cavity metastasis.
A 75-year-old woman presented with a painless swelling in the right mandibular retromolar area and numbness of the left lower lip. Radiographic examination of the mandible demonstrated an osteolytic lesion of the ascending ramus. Biopsy revealed adenocarcinoma of obscure origin. Staining of the specimen with a monoclonal antibody specific to colon carcinoma revealed its origin. On subsequent examinations, a primary tumor in the rectosigmoid region with extensive lung, liver and skeletal metastases were diagnosed. This unusual case of colonorectal carcinoma, presenting as a metastatic lesion of the mandible, was readily diagnosed by a novel immunohistochemical technique that utilizes highly specific monoclonal antibodies.


Does smoking among friends explain apparent genetic effects on current smoking in adolescence and young adulthood?
We used data from a prospective cohort study of twins to investigate the influence of unmeasured genetic and measured and unmeasured environmental factors on the smoking behaviour of adolescents and young adults. Twins were surveyed in 1988 (aged 11-18 years), 1991, 1996 and 2004 with data from 1409, 1121, 732 and 758 pairs analysed from each survey wave, respectively. Questionnaires assessed the smoking behaviour of twins and the perceived smoking behaviour of friends and parents. Using a novel logistic regression analysis, we simultaneously modelled individual risk and excess concordance for current smoking as a function of zygosity, survey wave, parental smoking and peer smoking. Being concordant for having peers who smoked was a predictor of concordance for current smoking (P<0.001). After adjusting for peer smoking, monozygotic (MZ) pairs were no more alike than dizygotic pairs for current smoking at waves 2, 3 and 4. Genetic explanations are not needed to explain the greater concordance for current smoking among adult MZ pairs. However, if they are invoked, the role of genes may be due to indirect effects acting through the social environment. Smoking prevention efforts may benefit more by targeting social factors than attempting to identify genetic factors associated with smoking.


Duodenal Follicular Lymphoma: Track or Treat?
Duodenal follicular lymphoma (DFL) is a rare variety of non-Hodgkin's lymphoma of the gastrointestinal tract that usually carries a favorable course, recognized as a new entity in 2016. It is usually diagnosed at an early stage located predominantly in the second portion of the duodenum. We report the case of a 74-year-old male patient with epigastric pain in whom gastroscopy revealed white mucosal nodules that were pathologically diagnosed as grade 1-2 DFL. Staging investigations revealed secondary lesions in the spleen and at the base of the tongue together with latero-cervical adenopathy. The tumor was stage IV according to the Lugano staging system. We reviewed the recent (last five years) literature defining the importance of combination therapy in the advanced stage. The patient achieved complete remission of the disease through chemoimmunotherapy following the Rituximab-Bendamustine scheme.


Characterization and gene deletion analysis of four homologues of group 3 pyridine nucleotide disulfide oxidoreductases from Thermococcus kodakarensis.
Enzymatic characterization of the four group 3 pyridine nucleotide disulfide oxidoreductase (PNDOR) homologues TK1299, TK0304, TK0828, and TK1481 from Thermococcus kodakarensis was performed, with a focus on their CoA-dependent elemental sulfur (S(0)) oxidoreductase (NSR) and NAD(P)H oxidase (NOX) activities. TK1299 exhibited NSR activity with a preference for NADPH and showed strict CoA-dependency similar to that of the Pyrococcus furiosus homologue PF1186. During the assays, the non-enzymatic formation of H2S from S(0) and free CoA-SH was observed, and the addition of enzyme and NADPH enhanced H2S evolution. A catalytic cycle of TK1299 was proposed suggesting that CoA-SH acted to solubilize S(0) by forming CoA persulfides, followed by reduction of an enzyme-S-S-CoA intermediate produced after both enzymatic and non-enzymatic evolution of H2S from the CoA persulfide, with NADPH as an electron donor. TK1481 showed NSR activity independently of CoA-SH, implying a direct reaction with S(0). TK1299, TK1481, and TK0304 exhibited high NOX activity, and the NADH-dependent activities were inhibited by the addition of free CoA-SH. Multiple disruptions of the four group 3 PNDOR homologues in T. kodakarensis demonstrated that none of these homologues were essential for S(0)-dependent growth. Many disruptants grew better than the parent strain, but a few multiple disruptants showed decreased growth properties after aerobic inoculation into a pyruvate-containing medium without S(0), suggesting the complicated participation of these group 3 PNDORs in sensitivity/resistance to dissolved oxygen when S(0) was absent.


Effects of sexual and physical abuse: a comparison of adolescent inpatients.
The present study was designed to collect descriptive data on a sample of hospitalized adolescents who were victims of sexual and/or physical abuse and to compare these groups to patients without a history of abuse. A questionnaire was administered to 51 inpatient teenagers at Fort Logan Mental Health Center, a state psychiatric hospital in Denver, Colorado. Information was obtained about family background, type, extent and duration of abuse, drug and alcohol abuse and self-reported mental health and general adjustment problems. In addition, clinical staff completed the Revised Behavior Problem Checklist (1) for the subjects. The results indicated statistically significant differences (p less than .05) between groups with adolescents who were both physically and sexually abused reporting the most problems. The implications for the findings for treatment are discussed.


DNASE1L3 inhibits hepatocellular carcinoma by delaying cell cycle progression through CDK2.
Dysregulated cell cycle targeting is a well-established therapeutic strategy against hepatocellular carcinoma (HCC). Dissecting the underlying mechanism may improve the efficacy of HCC therapy. HCC data from TCGA and new clinical samples were used for DNASE1L3 expression analysis and for assessing its correlation with HCC development. The in vitro function of DNASE1L3 in HCC cell proliferation, colony formation, migration and invasion was assessed using RTCA, CCK-8 and transwell assays and the in vivo function in subcutaneous tumor formation in a xenograft nude mouse model. The role of DNASE1L3 in HCC tumorigenesis was further verified in AKT/NRASV12-induced and DEN/CCl4-induced primary liver cancers in wildtype and Dnase1l3-/- mice. Finally, RNA-Seq analysis followed by biochemical methods including cell cycle, immunofluorescence, co-immunoprecipitation and Western blotting assays were employed to reveal the underlying mechanism. We found that DNASE1L3 was significantly downregulated and served as a favorable prognostic factor in HCC. DNASE1L3 dramatically attenuated HCC cell proliferation, colony formation, migration and invasion in vitro and reduced subcutaneous tumor formation in nude mice in vivo. Furthermore, DNASE1L3 overexpression dampened AKT/NRASV12-induced mouse liver cancer in wildtype mice and DNASE1L3 deficiency worsened DEN/CCl4-induced liver cancer in Dnase1l3-/- mice. Systemic analysis revealed that DNASE1L3 impaired HCC cell cycle progression by interacting with CDK2 and inhibiting CDK2-stimulated E2F1 activity. C-terminal deletion (DNASE1L3ΔCT) diminished the interaction with CDK2 and abrogated the inhibitory function against HCC. Our study unveils DNASE1L3 as a novel HCC cell cycle regulator and tumor suppressor. DNASE1L3 impairs HCC tumorigenesis by delaying cell cycle progression possibly through disrupting the positive E2F1-CDK2 regulatory loop. DNASE1L3 may serve as a target for the development of novel therapeutic strategies against HCC.


Removal of Protein-Bound Uremic Toxins by Liposome-Supported Peritoneal Dialysis.
Background:Protein-bound uremic toxins (PBUTs) are poorly cleared by peritoneal dialysis (PD). This study aimed to enhance PBUT removal in PD by adding a binder to the peritoneal dialysate and to evaluate the feasibility and efficacy of liposome-supported PD (LSPD) to increase the removal of PBUTs compared with albumin PD.Methods:Removal of p-cresyl sulfate (PCS), indoxyl sulfate (IS), and indole-3-acetic acid (3-IAA) was first evaluated in an in vitro PD model using artificial plasma preloaded with test solutes. Male Sprague-Dawley rats (n = 24) were then subjected to 5/6 nephrectomy and fed for 16 weeks to establish end-stage renal failure, after which they were treated with either conventional glucose-based PD, albumin-based PD, or liposome-based PD. Removal of PBUTs and small water-soluble solutes was determined during a 6-hour PD dwell.Results:In vitro experiments showed that adding albumin as a toxin binder to the dialysate markedly increased the removal of PCS, IS, and 3-IAA compared with the control. The uptake capacity of liposomes was comparable with that of albumin for PCS and 3-IAA, though slightly inferior for IS. In vivo PD in uremic rats demonstrated that LSPD resulted in higher intraperitoneal concentrations and more total mass removal for PBUTs than the conventional glucose-based PD, which was comparable with albumin PD.Conclusions:Supplementing conventional glucose-based PD solutions with a binder could efficiently increase the removal of PBUTs. This preliminary study suggested that LSPD may be a promising alternative to albumin PD for increasing PBUT removal in the development of next-generation PD solutions for PD patients.


Efficacy of tyrosine kinase inhibitors on a mouse chronic myeloid leukemia model and chronic myeloid leukemia stem cells.
Chronic myeloid leukemia (CML) is a hematopoietic stem cell disorder caused by constitutively active BCR-ABL1 tyrosine kinase resulting from the t(9;22) Philadelphia translocation. Imatinib, a BCR-ABL1 tyrosine kinase inhibitor (TKI), is a revolutionary molecular target inhibitor for CML. However, leukemic stem cells (LSCs) eventually become resistant to imatinib and thereby cause relapse. The next-generation BCR-ABL1 TKI dasatinib is also unable to eliminate CML LSCs. On the other hand, the third-generation BCR-ABL1 TKI ponatinib is not well studied in terms of its efficacy on CML LSCs. Here, we evaluate the efficacy of ponatinib against CML LSC-containing lin-Sca-1+c-Kit+ (LSK) cells using a mouse CML-like model. To this end, we compared the efficacy of imatinib, dasatinib, and ponatinib on CML LSK cells and showed that ponatinib is more effective at eliminating CML LSK cells. Our results suggest that ponatinib could be potentially useful for achieving treatment-free remission in CML patients.


Activation of Rubisco controls CO(2) assimilation in light: a perspective on its discovery.
CO(2) fixation during photosynthesis is regulated by the activity of ribulose bisphosphate carboxylase (Rubisco). This conclusion became more apparent to me after CO(2)-fixation experiments using isolated spinach chloroplasts and protoplasts, purified Rubisco enzyme, and intact leaves. Ribulose bisphosphate (RuBP) pools and activation of Rubisco were measured and compared to (14)CO(2) fixation in light. The rates of (14)CO (2) assimilation best followed the changes in Rubisco activation under moderate to high light intensities. RuBP pool sizes regulated (14) (2) assimilation only in very high CO(2) levels, low light and in darkness. Activation of Rubisco involves two separate processes: carbamylation of the protein and removal of inhibitors blocking carbamylation or blocking RuBP binding to carbamylated sites before reaction with CO(2) or O(2).


Post-traumatic stress and disability.
Post-traumatic stress is thought to account for significant disability. It is also known to be highly comorbid with other psychiatric conditions such as depression and alcohol dependence. To determine the relationship between post-traumatic stress, depression, alcohol dependence and disability. Seventy armed services personnel were assessed for DSM-IV diagnoses of post-traumatic stress disorder, major depressive disorder and alcohol dependence, and with continuous measures of symptoms of post-traumatic stress, depression and alcohol dependence following a traumatic event. These variables, as predictors of disability (using the Sheehan Disability Scale), were analysed using multivariate analysis of variance, analysis of covariance and multiple regression backward elimination models. No significant interaction was found for the diagnostic variables even after controlling for the continuous symptom measures. In the regression models, symptoms of depression were a significant predictor of total disability (R(2)=0.39). Symptoms of alcohol dependence and post-traumatic stress did not significantly predict disability. Since post-traumatic stress was not found to be associated with disability, its clinical importance may be questionable.


The inhibition of lipoprotein-associated phospholipase A2 exerts beneficial effects against atherosclerosis in LDLR-deficient mice.
To investigate the effects of darapladib, a specific inhibitor of lipoprotein-associated phospholipase A2 (lp-PLA2), on inflammation and atherosclerotic formation in the low density lipoprotein receptor (LDLR)-deficient mice. Six-week-old LDLR-deficient mice were fed an atherogenic high-fat diet for 17 weeks and then randomly divided into two groups. One group was administered darapladib (50 mg·kg(-1)·d(-1); po) for 6 weeks. The other group was administered saline as a control. Serum lipid levels were measured using the corresponding kits, and three inflammatory markers--interleukin-6 (IL-6), C reactive protein (hs-CRP), and platelet activating factor (PAF)--were determined using ELISA. Atherosclerotic plaque areas were stained with Sudan IV, and inflammatory gene expression at the lesions was evaluated using quantitative real-time PCR. The body weight and serum lipid level between the two groups were similar at the end of the dietary period. The serum lp-PLA2 activity, hs-CRP and IL-6 levels, however, were significantly reduced in the darpladib group. The inhibition of lp-PLA2 did not alter the serum PAF level. Furthermore, the plaque area, from the aortic arch to the abdominal aorta, was significantly reduced in the darpladib group. Additionally, the expression of inflammatory genes monocyte chemotactic protein-1 (MCP-1) and vascular cell adhesion molecule-1 (VCAM-1) was significantly reduced at the lesions in the darapladib group. Inhibition of lp-PLA2 by darapladib decreases the inflammatory burden and atherosclerotic plaque formation in LDLR-deficient mice, which may be a new strategy for the treatment of atherosclerosis.


Robust Virus-Specific Adaptive Immunity in COVID-19 Patients with SARS-CoV-2 Δ382 Variant Infection.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) that have become dominant as the pandemic progresses bear the ORF8 mutation together with multiple spike mutations. A 382-nucleotide deletion (Δ382) in the ORF7b and ORF8 regions has been associated with milder disease phenotype and less systemic inflammation in COVID-19 patients. However, its impact on host immunity against SARS-CoV-2 remains undefined. Here, RNA-sequencing was performed to elucidate whole blood transcriptomic profiles and identify contrasting immune signatures between patients infected with either wildtype or Δ382 SARS-CoV-2 variant. Interestingly, the immune landscape of Δ382 SARS-CoV-2 infected patients featured an increased adaptive immune response, evidenced by enrichment of genes related to T cell functionality, a more robust SARS-CoV-2-specific T cell immunity, as well as a more rapid antibody response. At the molecular level, eukaryotic initiation factor 2 signaling was found to be upregulated in patients bearing Δ382, and its associated genes were correlated with systemic levels of T cell-associated and pro-inflammatory cytokines. This study provides more in-depth insight into the host-pathogen interactions of ORF8 with great promise as a therapeutic target to combat SARS-CoV-2 infection.


Identification and purification of the main components of cellulases from a mutant strain of Trichoderma viride T 100-14.
A new mutant strain of fungus Trichoderma viride T 100-14 was cultivated on 1% microcrystalline cellulose (Avicel) for 120h and the resulting culture filtrate was prepared for protein identification and purification. To identify the predominant catalytic components, cellulases were separated by an adapted two-dimensional electrophoresis technique. The apparent major spots were identified by high performance liquid chromatography electrospray ionization mass (HPLC-ESI-MS). Seven of the components were previously known, i.e., the endoglucanases Cel7B (EG I), Cel12A (EG III), Cel61A (EG IV), the cellobiohydrolases Cel7A (CBH I), Cel6A (CBH II), Cel6B (CBH IIb) and the beta-glucosidase. The seven major components in the fermentation broth of T. viride T 100-14 probably constitute the essential enzymes for crystalline cellulose hydrolysis and they were further purified to electrophoretic homogeneity by a series of chromatography column. Hydrolysis studies of the purified elements revealed that three of the cellulases were classified as cellobiohydrolases due to their main activities on p-nitrophenyl-beta-d-cellobioside (pNPC). Three of the cellulases, with the abilities of hydrolyzing both carboxymethyl-cellulose (CMC) and Avicel indicate their endoglucanase activities. It deserved noting that the beta-glucosidase from the T 100-14 displayed an extremely high activity on p-nitrophenyl-beta-D-glycopyranoside (pNPG), which suggested it was a good candidate for the conversion of cellobiose to glucose.


Nuclear androgen receptors in the epithelium and stroma of human benign prostatic hypertrophic glands.
Androgen receptors have been characterized and quantified in nuclear extracts of separated epithelium and stroma from human benign prostatic hypertrophic (BPH) glands. Tritiated dihydrotestosterone was used as the ligand and incubation was carried out at 15 degrees C for 18-20 hr before separation of bound and free ligand using dextran-coated charcoal. The results were analysed by Scatchard-type analysis. The concentration of receptor was found to be significantly (p = 0.022) greater in stromal than in epithelial nuclei: 1765 +/- 152 vs 1030 +/- 227 fmol/mg DNA (SEM, n = 6). Fourteen competitors were tested and the results indicated the presence of specific androgen receptors rather than contaminating sex-hormone-binding globulin. This was also borne out by the results of agar gel electrophoresis and sucrose gradient ultracentrifugation studies. The results are in line with current opinion that prostatic stroma is an important androgen-sensitive tissue, particularly in human BPH.


Improving Effective Magnetic Resonance Imaging (MRI) Application in Soft Tissue Wrist Injury.
Magnetic resonance imaging (MRI) for soft-tissue wrist injury may be overprescribed, contributing to ineffective health care resource use. We aimed to discern predictive factors that may improve MRI's application in soft-tissue wrist injury. We conducted a retrospective chart review of adults who underwent MRIs for possible soft-tissue wrist injury between June 2009 and June 2014. Clinical data and treatment recommendations before and after MRI were analyzed. If the MRI-directed treatment recommendation was different from before MRI, the MRI was noted to have influenced the patient's treatment (Impact MRI). Among 140 MRI scans, 39 (28%) impacted treatment recommendation. Twenty-six Impact MRIs were ordered by hand surgeons, whereas 13 were ordered by referring physicians (P = .001). More Impact MRIs were found when an MRI was ordered for patients younger than 36 years (P = .01), within 6 weeks of symptom onset (P = .03), to question a specific anatomic injury (P = .0001), or by a board-certified hand surgeon (P = .001). Adjusting for other covariates, these 4 clinical factors were identified as independent predictive factors to Impact MRIs. MRIs for soft-tissue wrist injuries may more likely change management when the patient is younger, ordered within 6 weeks of symptom onset, and prescribed with a specific differential diagnosis. Referral to a hand surgeon should be considered before wrist MRI for the following patients: history of hand surgery/trauma, older than 36 years likely due to confounding chronic wrist changes, symptomatic for more than 6 weeks, and without clear differential diagnoses for the symptoms.


Quality-of-life impact after in-office treatment of nasal valve obstruction with a radiofrequency device: 2-year results from a multicenter, prospective clinical trial.
Insufficiency of the nasal valve is increasingly being recognized as a cause of nasal airway obstruction. The condition is associated with many symptoms, including nasal congestion, sleep disturbance, snoring, and an overall decline in quality of life (QoL). An in-office, minimally invasive radiofrequency treatment of the nasal valve has been associated with improved symptoms of nasal obstruction and patients' QoL for a 6-month period in a noncontrolled, prospective, single-arm study. The purpose of this study was to determine whether the results achieved with radiofrequency treatment at 6 months would be sustained through 24 months. Thirty-nine adult patients from an original cohort of 49 patients with severe to extreme Nasal Obstruction Symptom Evaluation (NOSE) Scale scores and dynamic or static internal nasal valve obstruction as the primary or significant contributor to obstruction were studied. Patients received intranasal bilateral radiofrequency treatment in a clinical study with a follow-up to 6 months, and were prospectively evaluated at 12, 18, and 24 months at 8 community-based otolaryngology practices. The patient-reported NOSE Scale score and 21 QoL questions were assessed. Clinically significant improvement from baseline in NOSE Scale score change demonstrated at 6 months (mean, 55.9; standard deviation [SD], 23.6; p < 0.0001) was maintained through 24 months (mean, 53.5; SD, 24.6; p < 0.0001). Responders (≥15-point improvement) consisted of 92.3% of participants at 6 months and 97.2% at 24 months. Responses to the QoL questions also showed improvement in patients' QoL. Treatment of the nasal valve with an in-office, transnasal temperature-controlled radiofrequency procedure was associated with stable and lasting improvement in symptoms of nasal obstruction and QoL through 24 months in this noncontrolled, single-arm study.


Optimizing therapy in primary biliary cholangitis: Alkaline phosphatase at six months identifies one-year non-responders and predicts survival.
Patients with primary biliary cholangitis (PBC) and insufficient response to ursodeoxycholic acid (UDCA), currently assessed after 1 year, are candidates for second-line therapy. The aims of this study are to assess biochemical response pattern and determine the utility of alkaline phosphatase (ALP) at six months as a predictor of insufficient response. UDCA-treated patients in the GLOBAL PBC database with available liver biochemistries at one year were included. POISE criteria were used to assess response to treatment, defined as ALP <1.67 × upper limit of normal (ULN) and normal total bilirubin at one year. Various thresholds of ALP at six months were evaluated to predict insufficient response based on negative predictive value (NPV) and that with nearest to 90% NPV was selected. For the study, 1362 patients were included, 1232 (90.5%) female, mean age of 54 years. The POISE criteria were met by 56.4% (n = 768) of patients at one year. The median ALP (IQR) of those who met POISE criteria compared to those who did not was 1.05 × ULN (0.82-1.33) vs. 2.37 × ULN (1.72-3.69) at six months (p < .001). Of 235 patients with serum ALP >1.9 × ULN at six months, 89% did not achieve POISE criteria (NPV) after one year of UDCA. Of those with insufficient response by POISE criteria at one year, 210 (67%) had an ALP >1.9 × ULN at six months and thus would have been identified early. We can identify patients for second-line therapy at six months using an ALP threshold of 1.9 × ULN, given that approximately 90% of these patients are non-responders according to POISE criteria.


Factors associated with career decision in Taiwanese nursing students: a questionnaire survey.
The vocational decisions of nursing students have always been a concern for nursing educators and administrators. It is unclear, however, what factors play a role in determining the professional choices made by these students. Limited research into this area has been undertaken in Taiwan. This study, therefore, was designed to investigate the factors associated with career choices in Taiwan's nursing students. The convenience sample included all fourth-year nursing students of a college in eastern Taiwan. A validated and reliable self-administered questionnaire developed by the investigators was used in the study. The sample population consisted of 231 fourth-year nursing students. Notably, 65.4% reported that they would not choose nursing as a career after graduation. Significant variables associated with this career decision were clinical ability (t=2.35, p<0.05), degree of stress during clinical practice (t=-3.04, p<0.01), and perceived support from staff nurses (t=2.28, p<0.05). These results suggest that both educators and administrators need to reconsider the way nursing students are educated, and indicate that strategies must be developed to enhance students' motivation to select nursing as a career.


Word repetition within- and across-visual fields: an event-related potential study.
A divided visual field (DVF) procedure was used to investigate the scalp distribution of the event-related potential (ERP) repetition effect. ERPs were recorded from 27 scalp sites whilst subjects (n = 20) discriminated between words and non-words presented to either the left (LVF) or the right (RVF) visual field. A proportion of the words were repeated on the trial immediately following their first presentation. In two within-field repetition conditions the two encounters with a word occurred in the same visual field (LVF or RVF). In two across-field repetition conditions, the two encounters with a word occurred in different visual fields. For both words and non-words, task performance was better for RVF presentations than for LVF presentations. In each repetition condition there was a positive-going shift in the ERP elicited by repeated words compared to that elicited by words on their first presentation. This ERP repetition effect was equivalent in magnitude and lateralised to the right hemisphere to an equivalent degree in all four repetition conditions. It is suggested that the ERP effects largely reflect the processing of visual form thought to occur predominately in the right hemisphere.


Factors influencing surgical management of proximal humerus fractures: do shoulder and trauma surgeons differ?
Proximal humerus fractures (PHFs) are managed with open reduction and internal fixation (ORIF), hemiarthroplasty (HA), reverse shoulder arthroplasty (RSA), or nonoperatively. Given the mixed results in the literature, the optimal treatment is unclear to surgeons. The purpose of this study was to survey orthopedic shoulder and trauma surgeons to identify the patient- and fracture-related characteristics that influence surgical decision-making. We distributed a 23-question closed-response email survey to members of the American Shoulder and Elbow Surgeons and Orthopaedic Trauma Association. Questions posed to respondents included demographics, surgical planning, indications for ORIF and arthroplasty, and the use of surgical augmentation with ORIF. Numerical and multiple-choice responses were compared between shoulder and trauma surgeons using unpaired t-tests and χ2 tests, respectively. Respondents included 172 shoulder and 78 trauma surgeons. When surgery is indicated, most shoulder and trauma surgeons treat 2-part (69%) and 3-part (53%) PHFs with ORIF. Indications for managing PHFs with arthroplasty instead of ORIF include an intra-articular fracture (82%), bone quality (76%), age (72%), and previous rotator cuff dysfunction (70%). In patients older than 50 years, 90% of respondents cited a head-split fracture as an indication for arthroplasty. Both shoulder and trauma surgeons preferred RSA for treating PHFs presenting with a head-split fracture in an elderly patient (94%), pre-existing rotator cuff tear (84%), and pre-existing glenohumeral arthritis with an intact cuff (75%). Similarly, both groups preferred ORIF for PHFs in young patients with a fracture dislocation (94%). In contrast, although most trauma surgeons preferred to manage PHFs in low functioning patients with a significantly displaced fracture or nonreconstructable injury nonoperatively (84% and 86%, respectively), shoulder surgeons preferred either RSA (44% and 46%, respectively) or nonoperative treatment (54% and 49%, respectively) (P < .001). Similarly, although trauma surgeons preferred to manage PHFs in young patients with a head-split fracture or limited humeral head subchondral bone with ORIF (98% and 87%, respectively), shoulder surgeons preferred either ORIF (54% and 62%, respectively) or HA (43% and 34%, respectively) (P < .001). ORIF and HA are preferred for treating simple PHFs in young patients with good bone quality or fracture dislocations, whereas RSA and nonoperative management are preferred for complex fractures in elderly patients with poor bone quality, rotator cuff dysfunction, or osteoarthritis. The preferred management differed between shoulder and trauma surgeons for half of the common PHF presentations, highlighting the need for future research.


Evidence for a diffusional model of Alzheimer amyloid A4 (beta-amyloid) deposition during neuritic plaque formation.
A recent study reported that Alzheimer senile plaques immunostained with monoclonal antibodies against the A4 (beta-amyloid) region of the amyloid precursor protein show gradients of density (Majocha R. E., Benes F. M., Reifel R. L., Rodenrys A. M. and Marotta C. A., Proc. natn. Acad. Sci. U.S.A. 85, 6182-6186, 1988). Although more than one explanation was suggested for this observation, the possible involvement of a diffusional process during plaque maturation was considered. In order to examine this hypothesis, specimens from prefrontal cortex, entorhinal area and hippocampal formation were immunoprocessed in a similar fashion and subjected to quantitative microdensitometric analyses of A4 amyloid reaction product. All plaques in the three brain areas examined showed a curvilinear relationship between the area of amyloid reaction product (expressed in pixel counts) and optical density (expressed as each of six grey scale levels). There was an increase in the area of amyloid at progressively lower density levels. When the area of amyloid reaction product at each density level was correlated with the overall size of individual plaques, it was found that there was a striking increase in the correlation coefficients at progressively lower grey scale levels, with r = 0.853 at the lowest level examined. When a second order derivation of these correlations was performed by expressing individual r-values with respect to an optical density index, an asymptotic relationship resulted with the lowest density levels showing an increasingly sharp rise toward unity. These data are consistent overall with a model for plaque maturation that involves diffusion of amyloid protein through the extracellular space from focal regions of high density where synthesis and/or release may occur.


Effect of environmental temperature and photoperiod on the melatonin levels in the pineal, lateral eye, and plasma of the frog, Rana perezi: importance of ocular melatonin.
Day-night melatonin concentrations were studied in the pineal body, lateral eye, and plasma of the frog Rana perezi in animals maintained in February and July under long (18L:6D) or short (6L:18D) photoperiod and high (25 +/- 1 degree) or low (6 +/- 1 degree) temperature in order to evaluate the influence of these environmental factors. When frogs were kept under short photoperiod and low temperature in February, no melatonin rhythm was observed in the pineal, ocular tissue, and plasma. High temperature at this period of the year induced a day-night rhythm of melatonin levels in the lateral eye and plasma. In July, under long photoperiod and high temperature, animals showed pronounced rhythms of melatonin in the pineal, eye, and plasma. A decrease of environmental temperature in this season abolished the melatonin rhythm. When animals were maintained in August under high (25 +/- 1 degree) temperature and long (18L:6D) or short (6L:18D) photoperiod, the duration of high night time ocular melatonin levels was correlated to the length of the dark phase. In all experiments the high ocular melatonin concentrations and the close parallelism observed between ocular and circulating melatonin profiles suggest that in this species melatonin could be released from the eyes in the general circulation.


A comprehensive and sensitive method for hair analysis in drug-facilitated crimes and incorporation of zolazepam and tiletamine into hair after a single exposure.
Hair is a highly relevant specimen that is used to verify drug exposure in victims of drug-facilitated crime (DFC) cases. In the present study, a new analytical method involving ultrahigh-performance liquid chromatography-tandem mass spectrometry was developed for determining the presence of model drugs, including zolazepam and tiletamine and their metabolites in hair specimens from DFCs. The incorporation of zolazepam and tiletamine into hair after a single exposure was investigated in Long-Evans rats with the ratio of the hair concentration to the area under the curve. For rapid and simple sample preparation, methanol extraction and protein precipitation were performed for hair and plasma, respectively. No interference was observed in drug-free hair or plasma, except for hair-derived diphenhydramine in blank hair. The coefficients of variance of the matrix effects were below 12%, and the recoveries of the analytes exceeded 70% in all of the matrices. The precision and accuracy results were satisfactory. The limits of quantification ranged from 20 to 50 pg in 10 mg of hair. The drug incorporation rates were 0.03 ± 0.01% for zolazepam and 2.09 ± 0.51% for tiletamine in pigmented hair. We applied the present method to real hair samples in order to determine the drug that was used in seven cases. These results suggest that this comprehensive and sensitive hair analysis method can successfully verify a drug after a single exposure in crimes and can be applied in forensic and clinical toxicology laboratories.


Ceramic Bearings with Titanium Adapter Sleeves Implanted During Revision Hip Arthroplasty Show Minimal Fretting or Corrosion: a Retrieval Analysis.
The BIOLOX® option system, consisting of a BIOLOX® delta ceramic femoral head with a titanium alloy adapter sleeve, is being increasingly utilized in revision hip arthroplasty. The sleeve protects the ceramic head from fracture and improper motion about the stem trunnion when a damaged trunnion is encountered at revision surgery. Corrosion and fretting due to metal-metal contact at the taper region of hip prosthesis create the potential of causing periprosthetic osteolysis and adverse local tissue reactions. The objective of this study was to identify the type and extent of damage to retrieved sleeves and ceramic heads to determine their in vivo performance. Twenty-four ceramic heads with titanium alloy sleeves were examined. The articular and taper surfaces for each ceramic head were assessed for metal transfer using a subjective grading system. All surfaces of the 24 titanium sleeves and stem trunnions (only available for 7 of 24 cases) were assessed for corrosion and fretting using an established grading system. Scanning electron microscopy and energy dispersive X-ray analysis were conducted on representative sample of sleeves. Fretting and corrosion were higher at the inner surface of the taper sleeve than the outer sleeve. Mean fretting scores at the inner taper and outer taper sleeve surfaces were 1.8 and 1.2, respectively. The mean corrosion score at the inner taper surface was 1.8; no corrosion was observed on the outer surface of any taper sleeve. SEM and EDS analyses provided further indications of low levels of damage. Fretting and corrosion were less severe than previously reported for conventional THA metal-metal taper connections, indicating that a ceramic head and titanium sleeve is a safe alternative in revision THA.


Ecology and Feeding Habits Drive Infection of Water Bugs with Mycobacterium ulcerans.
Mycobacterium ulcerans (MU), the causative agent of Buruli ulcer, is present in a wide spectrum of environments, including terrestrial and aquatic ecosystems in tropical regions. The most promising studies on the epidemiological risk of this disease suggest that some ecological settings may favor infection of animals with MU including human. A species' needs and impacts on resources and the environment, i.e., its ecological niche, may influence its susceptibility to be infected by this microbial form. For example, some Naucoridae may dive in fresh waters to prey upon infected animals and thus may get infected with MU. However, these studies have rarely considered that inference on the ecological settings favoring infection and transmission may be confounded because host carrier sister species have similar ecological niches, and potentially the same host-microbe interactions. Hence, a relationship between the ecological niche of Naucoridae and its infection with MU may be due to a symbiotic relationship between the host and the pathogen, rather than its ecological niche. To account for this confounding effect, we investigated the relationships between surrogates of the ecological niche of water bug species and their susceptibility to MU, by performing phylogenetic comparative analyses on a large dataset of 11 families of water bugs collected in 10 different sites across Cameroon, central Africa. Our results indicate that MU circulates and infects a couple of host taxa, i.e., Belostomatidae, Naucoridae, living both in the aquatic vegetation and as predators inside the trophic network and sister species of water bugs have indeed similar host-microbe interactions with MU.


Author Correction: Climate change, range shifts, and the disruption of a pollinator-plant complex.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Draft Genome Sequences of Prototrophic and Biotin-Auxotrophic Fusarium langsethiae Strains Isolated from an Oat Grain in the Northern Region of Russia.
Fusarium langsethiae is a suspected plant-pathogenic fungus causing cereal contamination with trichothecene mycotoxins. Here, we report the genome sequences of two F. langsethiae strains, MFG217701 (a prototroph) and MFG217702 (a biotin auxotroph), isolated from a grain of oat harvested in Russia.


New clinical forms of hereditary apoA-I amyloidosis entail both glomerular and retinal amyloidosis.
Apolipoprotein A1 amyloidosis (ApoAI) results from specific mutations in the APOA1 gene causing abnormal accumulation of amyloid fibrils in diverse tissues. The kidney is a prominent target tissue in ApoAI amyloidosis with a remarkable selectivity for the renal medulla. Here, we investigated six French families with ApoAI Glu34Lys, p.His179Profs∗47, and a novel p.Thr185Alafs∗41 variant revealing unprecedented clinical association of a glomerular with a retinal disease. Comprehensive clinicopathological, molecular and proteomics studies of numerous affected tissues ensured the correlation between clinical manifestations, including novel unrecognized phenotypes, and apoA-I amyloid deposition. These ophthalmic manifestations stemmed from apoA-I amyloid deposition, highlighting that the retina is a previously unrecognized tissue affected by ApoAI amyloidosis. Our study provides the first molecular evidence that a significant fraction of ApoAI amyloidosis cases with no family history result from spontaneous neomutations rather than variable disease penetrance. Finally, successful hepatorenal transplantation resulted in a life- and vision-saving measure for a 32-year-old man with a hitherto unreported severe ApoAI amyloidosis caused by the very rare Glu34Lys variant. Our findings reveal new modes of occurrence and expand the clinical spectrum of ApoAI amyloidosis. The awareness of glomerular and ocular manifestations in ApoAI amyloidosis should enable earlier diagnosis and avoid misdiagnosis with other forms of renal amyloidosis. Thus, documented apoA-I amyloid deposition in the retina offers new biological information about this disease and may change organ transplantation practice to reduce retinal damage in patients with ApoAI amyloidosis.


Blood pressure and blood pressure variability following withdrawal of propranolol and clonidine.
The effects of placebo, propranolol, and clonidine on blood pressure and blood pressure variability were examined in 14 subjects who had moderate essential hypertension. Hydrochlorothiazide was given throughout successive 4- to 5-week periods of placebo, propranolol, and clonidine administration. During each treatment period, subjects were admitted twice to the clinical research unit for 24-hour blood pressure monitoring performed during administration of placebo, propranolol and clonidine and repeated 1 to 2 weeks later during the first 24 hours after the abrupt cessation of placebo or drug administration. Blood pressure was recorded at 15-minute intervals using an automated noninvasive recorder (Arteriosonde, Roche, New Jersey). Systolic and diastolic blood pressure readings were averaged and the standard deviation taken as the measure of long-term variability (LTV). Systolic and diastolic blood pressure in sequential overlapping blocks of seven readings were averaged and the standard deviation calculated. Short-term variability (STV) was estimated as the average of the standard deviations of the running means. During placebo administration and withdrawal, systolic and diastolic blood pressures as well as LTV and STV were unchanged. Systolic and diastolic pressures did not differ during propranolol administration from those during propranolol withdrawal or placebo withdrawal. Systolic and diastolic LTV and STV did not differ during propranolol administration and withdrawal from those observed during placebo administration or withdrawal. After clonidine withdrawal, both systolic and diastolic pressures increased to values significantly greater than during clonidine administration. These values were significantly greater than those observed after placebo withdrawal, thereby indicating rebound hypertension. After clonidine withdrawal, seven of fourteen subjects developed systolic pressure rebound; diastolic pressure rebound was observed in three patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Myocardial adipose deposition and the development of heart failure with preserved ejection fraction.
It has been proposed that an increase of myocardial adiposity is related to left ventricular (LV) diastolic dysfunction. The specific roles of myocardial steatosis including epicardial fat and intramyocardial fat for diastolic function are unknown in those patients suffering heart failure (HF) with reduced (HFrEF) or preserved ejection fraction (HFpEF). This study aims to determine the complex relationship between myocardial adiposity in patients with HFrEF or HFpEF. Using cardiac magnetic resonance imaging (CMRI), myocardial steatosis was measured in 305 subjects (34 patients with HFrEF, 163 with HFpEF, and 108 non-HF controls). We also evaluated cardiac structure and diastolic and systolic function by echocardiography and CMRI. Patients with HFpEF had significantly more intramyocardial fat than HFrEF patients or non-HF controls [intramyocardial fat content (%), 1.56 (1.26, 1.89) vs. 0.75 (0.50, 0.87) and 1.0 (0.79, 1.15), P < 0.05]. Intramyocardial fat amount (%) was higher in HFpEF women than in men [1.91% (1.17%, 2.32%) vs. 1.22 (0.87%, 2.02%), P = 0.01]. When estimated by CMRI (left ventricular peak filling rate), echocardiographic E/e' level, or left atrial volume index, intramyocardial fat correlated with LV diastolic dysfunction parameters in HFpEF patients, and this was independent of age, co-morbidities, body mass index, gender, and myocardial fibrosis (standardized coefficient: β = -0.34, P = 0.03; β = 0.29, P = 0.025; and β = 0.25, P = 0.02, respectively). Patients with HFpEF had significantly more intramyocardial fat than HFrEF patients or non-HF controls. Independent of risk factors or gender, intramyocardial fat correlated with LV diastolic dysfunction parameters in HFpEF patients.


Characterization of the L-arginine:nitric oxide pathway in human platelets.
1. The activation of the L-arginine: nitric oxide (NO) pathway during aggregation of human platelets by adenosine 5'-diphosphate (ADP), arachidonic acid, thrombin and the calcium ionophore A23187 and its inhibition by NG-monomethyl-L-arginine (L-NMMA), NG-nitro-L-arginine methyl ester (L-NAME) and N-iminoethyl-L-ornithine (L-NIO) were studied. The inhibition of the cytosolic platelet NO synthase by these compounds was also examined. 2. Platelet aggregation induced by ADP (1-10 microM) and arachidonic acid (0.1-10 microM), but not that induced by thrombin (1-30 mu ml-1) or A23187 (1-10 nM), was inhibited by L-, but not D-arginine (1-30 microM). However, in the presence of a subthreshold concentration of prostacyclin (0.1 nM) or of M & B 22948 (1 microM), a selective inhibitor of guanosine 3':5'-cyclic monophosphate (cyclic GMP) phosphodiesterase, L-arginine caused concentration-dependent inhibition of aggregation induced by all of these aggregating agents. 3. L-NMMA, L-NAME and L-NIO (all at 1-30 microM), but not their D-enantiomers, enhanced to the same extent platelet aggregation induced by ADP, arachidonic acid and thrombin without affecting that induced by A23187. 4. In the presence of 300 microM L-arginine, the NO synthase in platelet cytosol was inhibited by L-NMMA, L-NAME and L-NIO with IC50s of 74 +/- 9, 79 +/- 8 and 8.5 +/- 1.5 microM (n = 3), respectively. 5. These results indicate that the L-arginine: NO pathway in human platelets plays a role in the modulation of platelet aggregation.


Late relapse of ulcerative colitis presenting as tracheobronchitis: a case report.
Lung involvement in inflammatory bowel diseases usually follows colitis. However, the time to lung involvement onset varies depending on the case, and pulmonary lesions are usually not parallel to exacerbations of the colitis. A 67-year-old Asian woman with a 38-year history of ulcerative colitis presented to our hospital with a complaint of prolonged dry cough for 2 months. The colitis had remained quiescent for > 35 years with low-dose salazosulfapyridine treatment. Chest computed tomography indicated circumferential thickening of the tracheal wall, while bronchoscopy examination revealed widespread erythematous edema and diffuse narrowing of the bronchial lumen. Biopsy of the bronchial mucosa showed submucosal lymphocytic infiltration. She was diagnosed with ulcerative-colitis-related tracheobronchitis and successfully treated with corticosteroids. Tracheobronchitis, in our case, occurred despite the longest remission period previously reported. Careful follow-up is necessary for the early recognition and treatment of pulmonary disease in patients with ulcerative colitis, regardless of the disease duration and long-term remission of colitis.


Organic matter content and particle size modifications in mangrove sediments as responses to sea level rise.
Mangroves sediments contain large reservoirs of organic material (OM) as mangrove ecosystems produce large quantities and rapidly burial OM. Sediment accumulation rates of approximately 2.0 mm year(-1), based on (210)Pb(ex) dating, were estimated at the margin of two well-developed mangrove forest in southern Brazil. Regional data point to a relative sea level (RSL) rise of up to ∼4.0 mm year(-1). This RSL rise in turn, may directly influence the origin and quantity of organic matter (OM) deposited along mangrove sediments. Lithostratigraphic changes show that sand deposition is replacing the mud (<63 μm) fraction and OM content is decreasing in successively younger sediments. Sediment accumulation in coastal areas that are not keeping pace with sea level rise is potentially conducive to the observed shifts in particle size and OM content.


Plasmodium falciparum Clinical Isolates Reveal Analogous Circulation of 3D7 and FC27 Allelic Variants and Multiplicity of Infection in Urban and Rural Settings: The Case of Adama and Its Surroundings, Oromia, Ethiopia.
Despite significant progress achieved globally in reducing malaria burden, still it is one of the major public health and economic problems in Ethiopia. Investigation of the local genetic polymorphism of P. falciparum, the most virulent and predominant malaria parasite primarily targeted in malaria control and elimination program, is paramount to assess intensity of parasite transmission. Analysis of the block 3 region of the msp-2 gene of P. falciparum provides strong molecular evidence to evaluate the real picture of malaria epidemiology to fine-tune the ongoing control and elimination programs in the region. Thus, this study was aimed at examining the status of such polymorphic gene and its implications in Adama and its surroundings. 148 isolates from patients with uncomplicated falciparum malaria were collected in the study from September 2019 to August 2020. Tween® 20 and the Chelex method were employed for parasite DNA extraction. msp-2 allelic families were genotyped by using nested polymerase chain reaction targeting its 3D7 and FC27 allelic variants followed by gel electrophoresis for fragment analysis. Seventeen different polymorphic forms of msp-2 allelic fragments were detected in the study area. Moreover, 47 (31.8%) and 41(27.7%) were detected for 3D7 and FC27 allelic families, respectively. Furthermore, the multiclonal allele type accounted for 60 (40.5%). The mean MOI was 1.4, and the heterogeneity index (He) is 0.49 indicating nearly intermediate malaria transmission in the study area. The study revealed nearly intermediate genetic diversity and mean MOI of P. falciparum in the study area, demanding further scale up of the ongoing control and elimination efforts.


[Traumatically induced junctional activity of nevocellular nevi (author's transl)].
Forty papillomatous nevi, mainly with junctional nests, were partially removed surgically in such a way that a small margin was left intact on one side and the deep nevus cells were retained on the flor of the wound. After healing of the wound, a small layer of scar tissue separated the regenerated epidermis from the deep nevus cells, which were left in the dermis. After very differing intervals, as early as 30 days after the partial surgical removal, typical alterations could be observed in the epidermis above the cicatrical tissue. Most frequently, there was a hyperpigmentation with an increase in the number of basal fluorescing dendritic cells, which could also be found along the uppermost parts of epidermal appendages. Moreover, nest-like conglomerations of pigment-producing cells appeared in the form of junctional nevus cell nests. About 2-3 month after the partial surgical removal "Abtropfung" was visible, indicating a regeneration of the nevus. Fluorescence-histochemical and enzyme-histochemical results revealed that marked alterations can be found not only in the newly formed epidermis above the scar, which morphologically show minor alterations.


Is CD69 an effective brake to control inflammatory diseases?
Early studies described CD69 as a leukocyte activation marker, and suggested its involvement in the activation of different leukocyte subsets as well as in the pathogenesis of chronic inflammation. However, recent investigations have showed that CD69 knockout mice exhibit an enhanced or reduced susceptibility to different experimental models of inflammatory diseases, including those mediated by T helper 17 (Th17) lymphocytes. In this regard, the expression of CD69, both in Th17 lymphocytes and by a subset of regulatory T cells, has an important role in the control of the immune response and the inflammatory phenomenon. Therefore, different evidence indicates that CD69 exerts a complex immunoregulatory role in humans, and that it could be considered as a target molecule for the therapy of immune-mediated diseases.


Tissue specificity of 8-prenylnaringenin: protection from ovariectomy induced bone loss with minimal trophic effects on the uterus.
Plant secondary metabolites with estrogenic activity (phyto-estrogens) have been studied in the past as a potential alternative to classical hormone-replacement therapy (HRT) in menopausal women. No final verdict on the efficacy of soy or red clover based pharmaceutical preparations has been reached despite numerous clinical studies. We have studied the novel and most potent phyto-estrogen 8-prenylnaringenin (8-PN) in adult ovariectomized rats, an established animal model to mimic hormone dependent osteoporosis in menopausal women. Our results demonstrate that 8-PN can completely protect from ovariectomy induced bone-loss while exhibiting minimal, (dose independent) trophic effects on uterus and endometrium. It is estimated that at equivalent bone protective doses of 17beta-estradiol and 8-PN, the phyto-estrogen has a 10-fold lower stimulatory effect on uterus and endometrium. The bone tissue specific effect of 8-PN was confirmed in a transgenic reporter mouse model (ERE-Luc mice). Here we also found pronounced estrogenic activity in prostate. Present results add important aspects to the pharmacological profile of 8-PN and position this compound as an interesting alternative new candidate for treatment of peri- and postmenopausal symptoms.


Residue-by-Residue View of In Vitro FUS Granules that Bind the C-Terminal Domain of RNA Polymerase II.
Phase-separated states of proteins underlie ribonucleoprotein (RNP) granules and nuclear RNA-binding protein assemblies that may nucleate protein inclusions associated with neurodegenerative diseases. We report that the N-terminal low-complexity domain of the RNA-binding protein Fused in Sarcoma (FUS LC) is structurally disordered and forms a liquid-like phase-separated state resembling RNP granules. This state directly binds the C-terminal domain of RNA polymerase II. Phase-separated FUS lacks static structures as probed by fluorescence microscopy, indicating they are distinct from both protein inclusions and hydrogels. We use solution nuclear magnetic resonance spectroscopy to directly probe the dynamic architecture within FUS liquid phase-separated assemblies. Importantly, we find that FUS LC retains disordered secondary structure even in the liquid phase-separated state. Therefore, we propose that disordered protein granules, even those made of aggregation-prone prion-like domains, are dynamic and disordered molecular assemblies with transiently formed protein-protein contacts.


Efficacy and safety of four COVID-19 vaccines in preventing SARS-CoV-2 infection: A rapid review.
Since the emergence of the SARS-CoV-2, there have been efforts to develop vaccines to control the COVID-19 pandemic. The present study assessed the efficacy and safety of the BNT162b2, mRNA-1273, ChAdOx1/AZD1222 and Gam-COVID-Vac rAd26-S/rAd5-S vaccines against the SARS-CoV-2. We searched PubMed/MEDLINE, Google Scholar, Cochrane, and the WHO International Clinical Trials Registry Platform on March 15, 2021. The search terms used were: "vaccine" OR "vaccination" AND "covid19" OR "coronavirus" OR "sarscov2" AND "bnt162b2" OR "chadox1-S" OR "azd1222" OR "sputnik" OR "Gam-COVID-Vac" OR "mrna" OR "mRNA-1273" . We measured the risk of bias of the studies and the quality of the evidence using GRADE profiles. A qualitative and quantitative analysis of the results of clinical trials is presented. Of the 74 identified studies, 4 were finally included in this review. The efficacies of the BNT162b2, mRNA-1273, ChAdOx1/AZD1222 and Gam-COVID-VacrAd26-S/rAd5-S vaccines against symptomatic COVID-19 were 95,0% (CI95% 90,3-97,6), 94,1% (CI95% 89,3-96,8), 66,7% (CI95% 57,4-74,0), and 91,1% (CI95% 83,8-95,1), respectively. There was moderate certainty of the evidence due to serious indirectness, when we measured the risk of bias of the studies and the quality of the evidence using GRADE profile. The safety profiles were acceptable, and data on serious adverse events (summary RR=0,93; CI95% 0,77-1,12; p=0,16) and deaths from all causes (summary RR=0,70; CI95% 0,33-1,50; p=0,90) showed no significant differences. The results of this review support the level of evidence for the efficacy and safety of the COVID-19 vaccines analysed.


[The three fundamental stages in the historical evolution of the surgical treatment of laryngeal cancer and the creators or authors of these stages].
This paper, written to honor the memory of one the greatest developers of ENT surgery, Professor R. Poch Viñals, presents a summary of what has been achieved in the surgical treatment of laryngeal carcinoma since the earlier attempts to eradicate this tumor up to the present time. These developments are written by someone who has lived and actively participated in the fight against this tumor from the time when really positive results were obtained up to nowadays. Currently, surgery still produces better results in the treatment of larynx malignancies than other therapeutic methods. However, we do not consider that the cancer cure results obtained with surgery are definitive. Therefore, physical and medical therapies should not be forgotten in our daily work since the latter--the only systemic treatment of cancer--is that in which we have our hopes.