EMA-real-time PCR as a reliable method for detection of viable Salmonella in chicken and eggs.
Culture-based Salmonella detection takes at least 4 d to complete. The use of TaqMan probes allows the real-time PCR technique to be a rapid and sensitive way to detect foodborne pathogens. However, unlike RNA-based PCR, DNA-based PCR techniques cannot differentiate between DNA from live and dead cells. Ethidium bromide monoazide (EMA) is a dye that can bind to DNA of dead cells and prevent its amplification by PCR. An EMA staining step prior to PCR allows for the effective inhibition of false positive results from DNA contamination by dead cells. The aim of this study was to design an accurate detection method that can detect only viable Salmonella cells from poultry products. The sensitivity of EMA staining coupled with real-time PCR was compared to that of an RNA-based reverse transcription (RT)-real-time PCR. To prevent false negative results, an internal amplification control was added to the same reaction mixture as the target Salmonella sequences. With an optimized EMA staining step, the detection range of a subsequent real-time PCR was determined to be 10(3) to 10(9) CFU/mL for pure cultures and 10(5) to 10(9) CFU/mL for food samples, which was a wider detection range than for RT-real-time PCR. After a 12-h enrichment step, EMA staining combined with real-time PCR could detect as low as 10 CFU/mL Salmonella from chicken rinses and egg broth. The use of EMA with a DNA-based real-time PCR can successfully prevent false positive results and represents a simple, yet accurate detection tool for enhancing the safety of food.


A new method of measuring propagation coefficients and characteristic impedance in blood vessels.
True propagation coefficients of pulse wave harmonics in an artery can be determined in vivo by measuring pulsatile blood pressure and flow at each of two points along the length of the vessel. These coefficients, which are complex numbers that describe the attenuation and the phase shift imposed on a traveling wave, are independent of the reflected waves in the circulation and thus provide information about the viscoelastic state and other properties of an artery. The equations involved are implicit in standard transmission-line theory, but they have not previously been applied in this particular way to blood vessels. The femoral artery, exposed in situ, was studied in 11 anesthetized dogs. At 1.5 Hz, true attenuation constants averaged 0.0151 nepers/cm, and true phase constants averaged 0.0155 radians/cm. As frequency increased, the apparent phase velocity of flow, in contrast, was relatively low at the first harmonic and rose as frequency increased. True phase velocities lay between the apparent pressure and flow values. Characteristic impedance at 1.5 Hz had an average modulus of 1.76 times 10-4 dyne sec/cm5 and a phase of minus 0.31 radians. The modulus diminished as frequency increased, and the phase became less negative. These results show that true phase constants and characteristic impedances determined by this method are consistent with data reported by others and provide information not previously available about flow wave propagation.


Mechanism of failure of recovery from hypoxic apnea by gasping in 17- to 23-day-old mice.
The mechanism of failure of autoresuscitation from hypoxic apnea in 17- to 23-day-old (weanling) Swiss Webster related/J mice was investigated by recording electrocardiogram (ECG) and ventilation in adult, weanling, and 11-day-old mice. Hypoxic apnea was induced with 97% N2-3% CO2. O2 (21% or 50% O2) or 97% N2-3% CO2 was given at the onset of apnea. The ECG showed no arrhythmias predictive of failure of autoresuscitation. The first indication of failure was a progressive fall in gasp volume ("run down"). This pattern also occurred in animals given continuous 97% N2-3% CO2 and was significantly different from that in mice that survived. Gasping duration in 97% N2 was longer in weanlings than adults but shorter than in 11 day olds. Respiratory and heart rate recovery were more rapid in adults than in weanlings. Although recovery in high O2 was more rapid, the survival rate was not increased. The lack of effect of high O2 on survival and the virtually identical pattern of gasping in mice dying in 97% N2 and air leads us to conclude that in mice that fail to autoresuscitate little or no O2 reaches the medullary respiratory centers. We speculate that this may be due to increased vulnerability of cardiac muscle to anoxia in 17- to 23-day-old mice, resulting in early and severe heart failure.


Spermidine and spermine delay brain aging by inducing autophagy in SAMP8 mice.
The natural polyamine spermidine and spermine have been reported to ameliorate aging and aging-induced dementia. However, the mechanism is still confused. An aging model, the senescence accelerated mouse-8 (SAMP8), was used in this study. Novel object recognition and the open field test results showed that oral administration of spermidine, spermine and rapamycin increased discrimination index, modified number, inner squares distance and times. Spermidine and spermine increased the activity of SOD, and decreased the level of MDA in the aging brain. Spermidine and spermine phosphorylate AMPK and regulate autophagy proteins (LC3, Beclin 1 and p62). Spermidine and spermine balanced mitochondrial and maintain energy for neuron, with the regulation of MFN1, MFN2, DRP1, COX IV and ATP. In addition, western blot results (Bcl-2, Bax and Caspase-3, NLRP3, IL-18, IL-1β) showed that spermidine and spermine prevented apoptosis and inflammation, and elevate the expression of neurotrophic factors, including NGF, PSD95and PSD93 and BDNF in neurons of SAMP8 mice. These results indicated that the effect of spermidine and spermine on anti-aging is related with improving autophagy and mitochondrial function.


Conduit-Free Retroperitoneal Access to the Iliac Artery in Endovascular Aortic Repair in Patients With Improper Access Vessels.
Successful endovascular aortic repair is highly dependent on the quality of the iliac access vessels. Patients with poor access vessels can be turned down from endovascular aortic repair or thoracic endovascular aortic repair by the treating physician. Perioperative complications such as failure to deliver the device or iliac rupture can be addressed to improper access vessels. In this article, we describe a novel technique to access the common iliac artery when access vessels are poor in diameter or quality. This sutureless conduit-free access technique can be used in TEVAR or EVAR and requires less surgical exposure of the iliac arteries.


[Valsalva minimal leak point pressure: a useful approximation to type III female urinary incontinence].
It has been reported that anatomic female urinary incontinence with complex sphincteric malposition can coexist with intrinsic damage of the sphincter itself. In this study we analyzed the utility of measuring minimum abdominal pressure at standardized bladder capacities that causes urinary incontinence in order to quantify intrinsic sphincteric damage in female urinary incontinence. The study comprised 50 women with urinary incontinence aged 36-78 years (mean 59.4), ICS standardized complete urodynamic study was performed. Minimum leak point pressure with Valsalva maneuver in decumbent and standing positions was determined during the filling phase of cystomanometry and it was defined as a measure of the abdominal pressure expressed as total baldder pressure without involuntary detrusor activity and exercised at standardized bladder capacities that originates objective urinary incontinence. Minimum leak point pressure for each bladder capacity was evaluated. Leak point pressures below 60 cm H2O indicate intrinsic sphincteric damage; pressures between 60 and 90 cm H2O indicate intrinsic damage and malposition or urethral hypermobility may coexist, and leak pressures over 90 cm H2O are related to complex sphincteric malposition. 5 women showed severe sphincteric deficiency (type III) and urinary incontinence was demonstrated with 50 ml bladder capacity and 30 cm H2O of abdominal pressure without detrusor activity. Thirty-five women (70%) had type II urinary incontinence. Of these, 10 (28.5%) showed intrinsic sphincteric damage in addition to malpositioning of the sphincteric complex at leak point pressures between 60 and 90 cm H2O. The rest of the women showed Blaivas' type 0 and I urinary incontinence. Valsalva minimum leak point pressure is a reproducible, reliable, useful and easily measured parameter in diagnosing female stress urinary incontinence. It allows approximation of the abdominal pressure to the level at which urinary leakage is produced during the filling phase of cystomanometry and gives us an idea of the extent of the intrinsic sphincteric damage, if any. Not only is sphincter damage demonstrated in type III urinary incontinence, but that it may also coexist to a varying degree with complex sphincteric malposition.


Impact of "basic life support & defibrillation" training on quality of telephone-assisted cardiopulmonary resuscitation.
While telephone-assisted cardiopulmonary resuscitation (T-CPR) is crucial for improving the chances of survival during cardiac arrest, there is limited information available on the effectiveness of T-CPR when administered by laypeople, especially those who are unfamiliar with these procedures. Therefore, we assessed the influence of basic life support and defibrillation (BLSD) training on the proficiency of T-CPR carried out by volunteer medical students participating in a BLSD course, using a simulated cardiac arrest scenario. The quality of T-CPR maneuvers was compared before and after the BLSD course. The results highlight the positive impact of BLSD training, significantly enhancing the quality of T-CPR and bringing it close to optimal levels.


DNA sequence determinants of lambda repressor binding in vivo.
The critical operator determinants for lambda repressor recognition have been defined by analyzing the binding of wild-type repressor to a set of mutant operators in vivo. Base pair substitutions at six positions within the lambda operator half-site impair binding severely, and define these base pairs as critical for operator function. One mutant operator binds repressor better than the consensus operator, and is a superoperator. The model proposed by M. Lewis in 1983 for the binding of lambda repressor to its operator accurately predicts the observed operator requirements for binding in vivo, with several minor exceptions. The order of affinities of the six natural lambda operators has also been determined.


Legislation, medicine, and politics: care for gender diverse youth.
A recent increase in legislation in the United States prohibiting gender-affirming care (GAC) for transgender youth follows a wave of its politicization despite support from all pertinent mainstream medical associations. This review describes the standards of GAC for transgender youth, the origins of legislation prohibiting this care, a review of current legislation in the United States and a discussion on the impact on patients, providers, and the medical field. A critical evaluation of historical parallels and current organizations supporting this legislation reveals it stems not from concerns within the medical field but from political and religious interests. This intrusion sets a dangerous precedent, undermining evidence-based medicine, providers' ability to practice according to standards of care, and patients' and guardians' autonomy and medical decision-making. This wave of antitrans rhetoric and legislation has resulted in threats to health providers and hospitals, 'moral distress" in providers, and migration of providers and patients from hostile states. Similar to antiabortion legislation, these legislative efforts will likely result in negative health outcomes and worsening disparities. The medical community must confront these forces directly through an understanding of the political and structural forces at play and adopting strategies to leverage collective power.


Data in question: A survey of European biobank professionals on ethical, legal and societal challenges of biobank research.
Biobanks have evolved, and their governance procedures have undergone important transformations. Our paper examines this issue by focusing on the perspective of the professionals working in management or scientific roles in research-based biobanks, who have an important impact on shaping these transformations. In particular, it highlights that recent advances in molecular medicine and genomic research have raised a range of ethical, legal and societal implications (ELSI) related to biobank-based research, impacting directly on regulations and local practices of informed consent (IC), private-public partnerships (PPPs), and engagement of participants. In our study, we investigate the ways that these concerns influence biobanking practices and assess the level of satisfaction of the cross-national biobanking research communities with the ELSI related procedures that are currently in place. We conducted an online survey among biobankers and researchers to investigate secondary use of data, informing and/or re-contacting participants, sharing of data with third parties from industry, participant engagement, and collaboration with industrial partners. Findings highlight the need for a more inclusive and transparent biobanking practice where biobanks are seen in a more active role in providing information and communicating with participants; the need to improve the current IC procedures and the role of biobanks in sharing of samples and data with industry partners and different countries, and the need for practical, tangible and hands-on ethical and legal guidance.


Minimally invasive anterolateral approaches for the treatment of back pain and adult degenerative deformity.
Minimally invasive and interbody and instrumented fusion techniques are increasingly being used for the treatment of adult degenerative disc disease, stenosis, and deformity of the lumbar spine. Advocates of minimal access spinal approaches list certain advantages over open procedures, including decreased postoperative pain and narcotic requirements, shorter hospital stays, less blood loss, and smaller incisions. The minimally invasive anterolateral approach allows access to the lumbar spine through the retroperitoneal space. We report on the short-term clinical and radiographic outcomes in four patients with mid to high lumbar coronal deformities treated at our institution with the anterolateral transpsoas minimally invasive approach. The primary presentation of these patients was back and leg pain. All patients showed improvement in their preoperative symptoms and solid arthrodesis at 6 months. Independent nonbiased patient pain analysis was also performed. Mean follow-up was 10 months (standard deviation, 1.4 mo), and mean hospital stay was 3.5 days (standard deviation, 1.9 d). One patient had additional posterior segmental instrumentation placed. Mean Cobb angles in the coronal plane were 28.5 degrees preoperatively and 18.3 degrees postoperatively (P < 0.05). We also present a historical perspective on retroperitoneal spine surgery, a regional anatomic description of the lumbosacral plexus and surrounding structures, and a description of the surgical technique as related to treatment of lumbar deformity.


Antiproliferative effects of GnRH agonists: prospects and problems for cancer therapy.
Gonadotropin-releasing hormone (GnRH) receptor activation has been demonstrated to inhibit cell proliferation in vitro and in vivo. These effects are dependent on the degree of receptor expression and the intracellular signaling protein milieu. The physiological and pathophysiological relevance is largely undefined, and its potential for exploitation in the treatment of specific malignancies is the subject of ongoing investigations. GnRH receptors are expressed in embryonic, juvenile and adult tissues, including brain, pituitary, gonads, accessory reproductive organs and placenta. The levels of receptor expression vary, from high in pituitary gonadotropes to low in peripheral tissues, although quantification of functional receptor protein has been determined in relatively few cell types. Roles for GnRH receptor signaling at different stages of animal development and its influence on reproductive health remain largely unexplored, except in cases of hereditary hypogonadal infertility. In addition to regulating hormone secretion, GnRH is postulated to act as a chemokine or a growth- and differentiation-inducing factor. Hence, receptor activation may influence the function of neuronal networks in the brain and the maturation of reproductive tissue epithelia. GnRH may also potentially influence the biology of cancerous cells in reproductive tissue since receptor activation may signal terminal differentiation, cell cycle arrest or apoptosis. In this context, the cell surface expression of GnRH receptor is important since it influences the intensity of intracellular signaling, and correlates with the ability to inhibit proliferation in transformed cells in vitro. Here, we review data on the effects of GnRH agonists on cell proliferation and apoptosis, and put forward hypotheses for investigation to determine whether the GnRH receptor acts as a tumor suppressor in neuroendocrine or epithelial cells.


Context change and associative learning.
This article reviews the effects of changing the background context on performance in associative learning tasks in humans and animals. The findings are complementary and consistent over animal conditioning (Pavlovian and instrumental learning) and human predictive learning and memory paradigms. In many cases, a context change after learning can have surprisingly little disruptive influence on performance. Extinction, or retroactive interference treatments more generally, is more context-specific than the initial learning. Contexts become important if the participant is exposed to any of several treatments that involve prediction error, which may serve to increase attention to the context. Contexts also become important if they are given predictive or informational value. Studies of instrumental (operant) learning are further consistent with the idea that the context might also influence affordances that support voluntary actions. Context switch effects are not universal, but mainly occur when certain attention and perception processes can come into play. WIREs Cogn Sci 2013, 4:237-244. doi: 10.1002/wcs.1225 This article is categorized under: Psychology > Learning.


A procedure to quantify the feed intake response of growing pigs to perturbations.
Improving robustness of farm animals is one of the goals in breeding programmes. However, robustness is a complex trait and not measurable directly. The objective of this study was to quantify and characterize (elements of) robustness in growing pigs. Robustness can be analysed by examining the animal's response to perturbations. Although the origin of perturbations may not be known, their effect on animal performance can be observed, for example, through changes in voluntary feed intake. A generic model and data analysis procedure was developed (1) to estimate the target trajectory of feed intake, which is the amount of feed that a pig desires to eat when it is not facing any perturbations; (2) to detect potential perturbations, which are deviations of feed intake from the estimated target trajectory; and (3) to characterize and quantify the response of the growing pigs to the perturbations using voluntary feed intake as response criterion. The response of a pig to a perturbation is characterized by four parameters. The start and end times of the perturbation are 'imposed' by the perturbing factor, while two other parameters describe the resistance and resilience potential of the pig. One of these describes the immediate reduction in daily feed intake at the start of the perturbation (i.e., a 'resistance' trait) while another parameter describes the capacity of the pig to adapt to the perturbation through compensatory feed intake to rejoin the target trajectory of feed intake (i.e., a 'resilience' trait). The procedure has been employed successfully to identify the target trajectory of feed intake in growing pigs and to quantify the pig's response to a perturbation.


Functional state following the Fontan procedure.
Despite improvements in outcomes after completion of the Fontan circulation, long-term functional state varies. We sought to identify pre- and postoperative characteristics associated with overall function. We analyzed data from 476 survivors with the Fontan circulation enrolled in the Pediatric Heart Network Fontan Cross-sectional Study. Mean age at creation of the Fontan circulation was 3.4 plus or minus 2.1 years, with a range from 0.7 to 17.5 years, and time since completion was 8.7 plus or minus 3.4 years, the range being from 1.1 to 17.3 years. We calculated a functional score for the survivors by averaging the percentile ranks of ventricular ejection fraction, maximal consumption of oxygen, the physical summary score for the Child Health Questionnaire, and a function of brain natriuretic peptide. The mean calculated score was 49.5 plus or minus 17.3, with a range from 3 to 87. After adjustment for time since completion of the circulation, we found that a lower score, and hence worse functional state, was associated with: right ventricular morphology (p less than 0.001), higher ventricular end-diastolic pressure (p equals 0.003) and lower saturations of oxygen (p equals 0.047) prior to completion of the Fontan circulation, lower income for the caregiver (p equals 0.003), and, in subjects without a prior superior cavopulmonary anastomosis, arrhythmias after completion of the circulation (p equals 0.003). The model explained almost one-fifth (18%) of the variation in the calculated scores. The score was not associated with surgical centre, sex, age, weight, fenestration, or the period of stay in hospital after completion of the Fontan circuit. A validation model, using 71 subjects randomly excluded from initial analysis, weakly correlated (R equals 0.17, p equals 0.16) with the score calculated from the dataset. Right ventricular morphology, higher ventricular end-diastolic pressure and lower saturations of oxygen prior to completion of the Fontan circuit, lower income for the provider of care, and arrhythmias after creation of the circuit, are all associated with a worse functional state. Unmeasured factors also influence outcomes.


Mechanism of increased alpha adrenergic vasoconstriction in human essential hypertension.
Multiple components of vascular alpha adrenergic responsiveness were investigated in twenty-four men with mild hypertension and eighteen age- and weight-matched normotensive controls. Arterial plasma norepinephrine (paNE), an index of sympathetic drive, was increased in hypertensives compared to normotensives (mean +/- SE), 199 +/- 24 vs. 134 +/- 11 pg/ml, P less than 0.02. The effective concentration of intra-arterial (iaNE) increasing forearm vascular resistance (FAVR) 30% (NE-EC30, an index of vascular alpha-receptor sensitivity) was similar in normotensives and hypertensives, 9 +/- 1 vs. 13 +/- 3 ng/100 ml per min, respectively, P greater than 0.3. The phentolamine induced reduction in FAVR, an index of vascular alpha-tone, was greater in hypertensives, -21.3 +/- 1.8 vs. normotensives, -14.9 +/- 1.2 U, P less than 0.02. We interpret these data as evidence for normal vascular alpha-receptor sensitivity to norepinephrine in mild hypertensives. Consequently, the increased sympathetic drive in mild hypertensives explains the elevated vascular alpha-tone. Although vascular alpha-receptor sensitivity to iaNE was normal, the FAVR responses at high doses (reactivity) were greater in hypertensives to regional infusion of both NE and angiotensin II. This "nonspecific" enhancement of vascular reactivity is probably explained by structural vascular changes in hypertensives.


Artificial intelligence based de-novo design for novel Plasmodium falciparum plasmepsin (PM) X inhibitors.
Plasmodium falciparum is the leading cause of malaria with 627,000 deaths annually. Invasion and egress are critical stages for successful infection of the host yet depend on proteins that are extensively pre-processed by various maturases. Plasmepsins (Plasmodium pepsins, abbreviated PM, I-X) are pepsin-like aspartic proteases that are involved in almost all stages of the life cycle. The goal of this study was to use de-novo generative modeling techniques to create novel potential PfPMX inhibitors. A total of 4325 compounds were virtually screened by structural-based docking methods. The obtained hits were utilized to refine a structure-based Ligand Neural Network (L-Net) generative model to generate related compounds. The obtained optimal L-Net Compounds with smina scores ≤ -5.00KCalmol-1 and QED ≥ 0.35 were further taken for amplification utilizing Ligand Based Transformer modeling using Deep generative learning (Drug Explorer/DrugEx). The resulting hits were then subjected to XP Glide conventional Molecular docking and QikProp ADMET screening; molecules with XP Docking score ≤ -7.00KCalmol-1 were retained. Based on their Glide ligand efficiency, originality, and uniqueness, 30 compounds were chosen for binding affinity and MM_GBSA energy determination. Following Induced Fit docking (IFD), 7 compounds were taken for 50 ns MD simulations and FEP/MD calculations. This study reported novel potential PfPMX inhibitors with acceptable ADMET profiles and reasonable synthetic accessibility scores, as well as sufficient docking scores against other PMs were generated. The PfPMX inhibitors reported in this article are promising antimalarials for the next stages of drug development, and the first of their kind to be investigated thoroughly.Communicated by Ramaswamy H. Sarma.


Preoperative infusional chemoradiation, selective intraoperative radiation, and resection for locally advanced pelvic recurrence of colorectal adenocarcinoma.
The results of preoperative infusional chemoradiation, resection, and selective intraoperative radiation (IORT) boost in 43 previously nonirradiated patients with locally advanced pelvic recurrence of colorectal adenocarcinoma are described. After surgery alone 10% to 30% of patients with carcinoma of the distal colon and rectum will develop isolated pelvic recurrence. In most cases, the disease is locally advanced and not amenable to curative resection. Preoperative infusional chemoradiation has been shown to increase resectability and decrease local recurrence in primary locally advanced colorectal cancer. Based on this experience, we initiated a multimodality treatment protocol to treat patients with pelvic recurrence of colorectal adenocarcinoma. Forty-three consecutive patients with histologically proven pelvic recurrence of colorectal adenocarcinoma were enrolled on a multimodality treatment protocol. The treatment plan consisted of 5 weeks of concurrent pelvic external beam radiotherapy (45 Gy) with continuous intravenous infusion of 5-fluorouracil and/or cisplatin. This was followed by surgery that included IORT boost (10-20 Gy) for 21 patients and brachytherapy for 4 patients. Forty patients (93%) underwent operation and 33 (77%) underwent resection with curative intent. There were 29 (88%) margin-negative resections. Fifteen patients (48%) underwent sphincter-preserving operations. There were no treatment-related deaths. Twenty-two patients experience perioperative complications. Median follow-up for the 43 patients was 26 months. The local recurrence rate was 36%. Median survival for the patients who underwent resection was 34 months, and actuarial 5-year disease-free and overall survival were 37% and 58%, respectively. Tumor cytoreduction by preoperative chemoradiation can increase resectability and enable sphincter-preserving surgery in patients with locally advanced pelvic recurrence of colorectal cancer.


Small intestinal submucosa for vascular reconstruction in the presence of gastrointestinal contamination.
Surgical options for vascular reconstruction in a contaminated field are limited and include prosthetic reconstruction or ligation with extra-anatomic bypass. With prosthetic insertion, rates of graft infection and failures (pseudoaneurysms and thrombosis) are high. In the emergent situations, extra-anatomic bypass is time-consuming and complex, and it produces marginal long-term results. Small intestinal submucosa (SIS) is a cell-free collagen matrix derived from porcine small intestine. Preliminary studies have demonstrated its ability to be remodeled into host tissue. In this study, we compared SIS to polytetrafluoroethylene (PTFE) as a vascular patch for arterial repair in the presence of massive gastrointestinal contamination to evaluate graft patency, incorporation, infection, and aneurysm formation. Adult mongrel pigs underwent general anesthesia with Isoflurane and were then randomized to 1 of 3 groups: control, contamination (colon puncture with stool contamination of the pelvis), or shock + contamination (40% blood volume for 1 hour, then resuscitation with shed blood and crystalloid, plus contamination). All groups then underwent a left common iliac arteriotomy and further randomized to a 1 x 3-cm patch angioplasty with either SIS or PTFE. All received cefotetan for 24 hours. All animals were sacrificed between 2 and 4 weeks, and necropsy was performed. Grafts were cultured, and microscopic analysis with hematoxylin and eosin and trichrome was performed. Outcomes included pulse quality (normal or diminished) compared with opposite side, graft infection, and pseudoaneurysm; all were determined by a blinded investigator. Forty animals were randomized, and 1 died of abdominal sepsis. All control animals had normal distal pulses, no pseudoaneurysms, and no patch infections. The pseudoaneurysm rate for the contaminated PTFE patches was 25% compared with 0% in the SIS group (P = 0.09). Patch infection occurred in 73% of all PTFE patches compared with 8% of SIS patches (P < 0.03). Organisms present in the infected grafts included Escherichia coli, Bacteroides species, and other Gram-negative enterics. Histopathology demonstrated the presence of neointima in both SIS and PTFE. Only SIS was completely incorporated, with infiltration of collagen fibrils and lymphocytes. SIS was associated with improved graft patency, less infection, complete incorporation, and no false aneurysm formation when compared with PTFE. This may be due to its ability to provide a durable scaffold for cellularization and tissue remodeling. This material may offer a superior alternative to more complex vascular reconstruction techniques in contaminated fields.


Consolation as possible expression of sympathetic concern among chimpanzees.
Chimpanzees are known to spontaneously provide contact comfort to recent victims of aggression, a behavior known as consolation. Similar behavior in human children is attributed to empathic or sympathetic concern. In line with this empathy hypothesis, chimpanzee consolation has been shown to reduce the recipient's state of arousal, hence to likely alleviate distress. Other predictions from the empathy hypothesis have rarely been tested, however, owing to small sample sizes in previous studies. An exceptionally large database of spontaneous consolation in two outdoor-housed groups of chimpanzees lends further support to the empathy hypothesis in that consolation occurred disproportionally between individuals that are socially close (i.e., kin and affiliation partners) and was more typical of females than males, which differences are also known of human empathy. These effects were demonstrated using generalized linear mixed models, which control multiple variables at once. An exception to the above pattern was formed by the highest-ranking males, which frequently offered consolation to victims of aggression, probably as part of their general policing function in chimpanzee society. Consolation occurred more frequently in the absence of reconciliation between former opponents, suggesting that actors are sensitive to the contact need of victims of aggression, which may be greater if the aggressor ignores them. That consolation is an integrated part of close mutual relationships is supported by the tendency for it being reciprocated.


Gastrointestinal tumorigenesis in Smad4 (Dpc4) mutant mice.
The SMAD4 (Dpc4) gene plays a key role in the TGF-beta signaling pathway. We recently inactivated the mouse homolog Smad4. The homozygous mutants were embryonic lethals, whereas the heterozygotes were viable and fertile. Although young heterozygotes were normal, old mice developed gastric and duodenal polyps similar to those found in human juvenile polyps characterized by abundant stroma and eosinophilic infiltrations. These data are consistent with the reports that a subset of human juvenile polyposis kindreds carry germline mutations in the SMAD4 gene. We then introduced the Smad4 mutation into the Apc delta 716 knockout mice, a model for human familial adenomatous polyposis. Because both Apc and Smad4 are located on mouse chromosome 18, we constructed by meiotic recombination, compound heterozygotes carrying both mutations on the same chromosome. In such mice, intestinal polyps developed into more malignant tumors than those in the simple Apc delta 716 heterozygotes, showing an extensive stromal cell proliferation and strong submucosal invasion. These results indicate that mutations in SMAD4 play a significant role in the malignant progression of colorectal tumors.


Fentanyl vapor self-administration model in mice to study opioid addiction.
Intravenous drug self-administration is considered the "gold standard" model to investigate the neurobiology of drug addiction in rodents. However, its use in mice is limited by frequent complications of intravenous catheterization. Given the many advantages of using mice in biomedical research, we developed a noninvasive mouse model of opioid self-administration using vaporized fentanyl. Mice readily self-administered fentanyl vapor, titrated their drug intake, and exhibited addiction-like behaviors, including escalation of drug intake, somatic signs of withdrawal, drug intake despite punishment, and reinstatement of drug seeking. Electrophysiological recordings from ventral tegmental area dopamine neurons showed a lower amplitude of GABAB receptor-dependent currents during protracted abstinence from fentanyl vapor self-administration. This mouse model of fentanyl self-administration recapitulates key features of opioid addiction, overcomes limitations of the intravenous model, and allows investigation of the neurobiology of opioid addiction in unprecedented ways.


Complications of endoscopic third ventriculostomy in previously shunted patients.
Endoscopic third ventriculostomy (ETV) is considered to be a safe and effective treatment in selected patients as an initial treatment for obstructive hydrocephalus and at the time of shunt malfunction in previously shunted patients. We compared the outcome and complications of ETV between patients with newly diagnosed hydrocephalus and those with previous shunting procedures. A retrospective review of patients undergoing ETV from 1996 to 2004 at Alberta's Childrens Hospital and Foothills Medical Centre was completed. Patient data included symptoms at clinical presentation, cause of hydrocephalus, age at initial shunt, number of previous shunt revisions, age at ETV, complications, and subsequent shunting procedures performed. A total of 131 patients were identified with a minimum follow-up duration of 1 year; 71 (82.5%) of 86 patients who underwent ETV as a primary procedure and 36 (80%) of 45 patients who had ETV at the time of shunt malfunction were shunt-free at the last follow-up evaluation. Patients younger than 1 year old who underwent ETV were more likely to require an additional procedure for control of their hydrocephalus (P < 0.01). Serious complications after ETV occurred more frequently in patients who presented at the time of shunt malfunction (14 of 45 patients, 31%) compared with patients who underwent primary ETV (seven of 86 patients, 8%) (P = 0.02). Previously shunted patients with a history of two or more revisions (P = 0.03) and who experienced a serious complication at the time of ETV (P = 0.01) were more likely to require shunt replacement. ETV is an effective treatment both in selected patients with newly diagnosed hydrocephalus and in patients with a previous shunting procedure who are presenting with malfunction. Complications of ETV occur more frequently in previously shunted patients than in patients treated for newly diagnosed hydrocephalus, and care must be taken in the selection and treatment of these patients.


Caregiving experiences of family members of persons with dementia in south India.
This study reports on the first phase of an investigation aimed at adapting The Savvy Caregiver program, a successful family caregiving curriculum developed in the United States, for application in South India. Thirty family members caring for a person with dementia were interviewed regarding their experiences as caregivers (CGs). Qualitative interviews were conducted with the family member at a geriatric clinic, while other diagnostic procedures were being carried out with the person with dementia. Findings from the study revealed that although family members understood the term CG, none could identify a word for CG in his or her language. There was little understanding of dementia as an illness. Family CGs reported feeling distressed, overwhelmed, and frustrated with caregiving. Caregivers were interested in an educational program, but many had unrealistic expectations for what they wanted to learn. The findings provide directions for adapting The Savvy Caregiver curriculum for Indian family CGs.


Correlated fluctuations carry signatures of gene regulatory network dynamics.
The dynamics of transcriptional control involve small numbers of molecules and result in significant fluctuations in protein and mRNA concentrations. The correlations between these intrinsic fluctuations then offer, via the fluctuation dissipation relation, the possibility of capturing the system's response to external perturbations, and hence the nature of the regulatory activity itself. We show that for simple regulatory networks of activators and repressors, the correlated fluctuations between molecular species show distinct characteristics for changes in regulatory mechanism and for changes to the topology of causal influence. Here, we do a stochastic analysis and derive time-dependent correlation functions between molecular species of regulatory networks and present analytical and numerical results on peaks and delays in correlations between proteins within networks. Upon using these values of peaks and delays as a two-dimensional feature space, we find that different regulatory mechanisms separate into distinct clusters. This indicates that experimentally observable pairwise correlations can distinguish between gene regulatory networks.


Leukocyte esterase-nitrite and bioluminescence assays as urine screens.
The 1-min leukocyte esterase (LE)-nitrite test (Chemstrip 9; Biodynamics, Division of Boehringer Mannheim Biochemicals, Indianapolis, Ind.) and a bioluminescence assay (Monolight centrifugation method; Analytical Luminescence Laboratory, Inc., San Diego, Calif.) were tested for their efficacy as urine screens among 453 patients at a tertiary-care teaching hospital. Both methods had the capacity to exclude significant bacteriuria (greater than or equal to 10(5) CFU/ml) when compared with the results of conventional culture methods, with predictive values of 99 and 93%, respectively, for a negative test. Bioluminescence was the more accurate nonculture method used. Sensitivity and specificity values were 97 and 71%, respectively, for bioluminescence, 82 and 60%, respectively, for LE with nitrite, and 72 and 64%, respectively, for LE without nitrite. At reduced levels of bacteriuria less than 10(5) CFU/ml), the sensitivities of LE-nitrite and bioluminescence were decreased but comparable. The addition of protein and blood test results in the Chemstrip 9, along with LE-nitrite as bacteriuria indicators, were unsatisfactory because of the large numbers of false-positive results attributed to protein and blood determinations. LE activity as detected by the LE test was a poor predictor of significant bacteriuria in both male and female patients. The sensitivity (71%) and specificity (57%) of the LE test in male patients were significantly lower than those previously reported and varied with the patient population studied.


Dicer Regulates the Balance of Short-Lived Effector and Long-Lived Memory CD8 T Cell Lineages.
MicroRNAs constitute a major post-transcriptional mechanism for controlling protein expression, and are emerging as key regulators during T cell development and function. Recent reports of augmented CD8 T cell activation and effector differentiation, and aberrant migratory properties upon ablation of Dicer/miRNAs in naïve cells have established a regulatory role of miRNAs during priming. Whether miRNAs continue to exert similar functions or are dispensable during later stages of CD8 T cell expansion and memory differentiation remains unclear. Here, we report a critical role of Dicer/miRNAs in regulating the balance of long-lived memory and short-lived terminal effector fates during the post-priming stages when CD8 T cells undergo clonal expansion to generate a large cytotoxic T lymphocyte (CTL) pool and subsequently differentiate into a quiescent memory state. Conditional ablation of Dicer/miRNAs in early effector CD8 T cells following optimal activation and expression of granzyme B, using unique dicerfl/fl gzmb-cre mice, led to a strikingly diminished peak effector size relative to wild-type antigen-specific cells in the same infectious milieu. Diminished expansion of Dicer-ablated CD8 T cells was associated with lack of sustained antigen-driven proliferation and reduced accumulation of short-lived effector cells. Additionally, Dicer-ablated CD8 T cells exhibited more pronounced contraction after pathogen clearance and comprised a significantly smaller proportion of the memory pool, despite significantly higher proportions of CD127Hi memory precursors at the effector peak. Combined with previous reports of dynamic changes in miRNA expression as CD8 T cells differentiate from naïve to effector and memory states, these findings support distinct stage-specific roles of miRNA-dependent gene regulation during CD8 T cell differentiation.


A network model of transient polymers: exploring the micromechanics of nonlinear viscoelasticity.
Dynamic networks contain crosslinks that re-associate after disconnecting, imparting them with viscoelastic properties. While continuum approaches have been developed to analyze their mechanical response, these approaches can only describe their evolution in an average sense, omitting local, stochastic mechanisms that are critical to damage initiation or strain localization. To address these limitations, we introduce a discrete numerical model that mesoscopically coarse-grains the individual constituents of a dynamic network to predict its mechanical and topological evolution. Each constituent consists of a set of flexible chains that are permanently cross-linked at one end and contain reversible binding sites at their free ends. We incorporate nonlinear force-extension of individual chains via a Langevin model, slip-bond dissociation through Eyring's model, and spatiotemporally-dependent bond attachment based on scaling theory. Applying incompressible, uniaxial tension to representative volume elements at a range of constant strain rates and network connectivities, we then compare the mechanical response of these networks to that predicted by the transient network theory. Ultimately, we find that the idealized continuum approach remains suitable for networks with high chain concentrations when deformed at low strain rates, yet the mesoscale model proves necessary for the exploration of localized stochastic events, such as variability of the bond kinetics, or the nucleation of micro-cavities that likely conceive damage and fracture.


Using a coupled LES aerosol-radiation model to investigate the importance of aerosol-boundary layer feedback in a Beijing haze episode.
Haze episodes, characterised by extremely high aerosol concentrations and a reduction in visibility to less than 10 km, are a frequent occurrence in wintertime Beijing, despite policy interventions leading to an overall improvement in average annual air quality. The main drivers in the onset of haze episodes in wintertime Beijing are changing synoptic conditions, however, aerosol-radiation interactions and their feedback on boundary layer meteorology are thought to play an essential role in the intensity and longevity of haze episodes. In this study we use a coupled LES aerosol-radiation model (UCLALES-SALSA), which we have recently configured for the urban environment of Beijing. The model's high resolution and control over meteorological and aerosol conditions as well as atmospheric processes means we can directly elucidate and quantify the importance of specific aspects of the aerosol-radiation-meteorology feedback in the cumulative stage of Beijing haze. The main results presented here show (a) synoptic scale meteorology has a larger impact on boundary layer suppression than high aerosol concentrations and (b) unlike previous results obtained using regional models or observationally driven analyses, there is no threshold value at which the aerosol-radiation-meteorology feedback has a significant effect on PBL height. Rather, our work shows that for the aerosol composition in this case study, the role of the feedback effect in reducing PBL height increases under shallow boundary layer conditions and with increasing pollution loading in an almost linear fashion. This lack of a threshold found for our case study has important policy implications since interventions based on such a value will not result in large reductions associated with turning off the feedback process. Furthermore, this work directly shows that although the right synoptic changes are a prerequisite for pollution episodes in Beijing, local and regional emissions drive increases in aerosol load that are sufficient to initiate the aerosol feedback loop. This further drives suppression of the boundary layer top and promotes stagnation of air and increased stability, which can be self-sustaining. This results in higher surface aerosol concentrations for extended periods of time, with severe consequences for human health [Lv et al., Atmos. Environ., 2016, 124, 98-108; Wang et al., Atmos. Chem. Phys., 2019, 19(10), 6949-6967].


Competition of individual domain folding with inter-domain interaction in WW domain engineered repeat proteins.
Engineered repeat proteins have proven to be a fertile ground for studying the competition between folding, misfolding and transient aggregation of tethered protein domains. We examine the interplay between folding and inter-domain interactions of engineered FiP35 WW domain repeat proteins with n = 1 through 5 repeats. We characterize protein expression, thermal and guanidium melts, as well as laser T-jump kinetics. All experimental data is fitted by a global fitting model with two states per domain (U, N), plus a third state M to account for non-native states due to domain interactions present in all but the monomer. A detailed structural model is provided by coarse-grained simulated annealing using the AWSEM Hamiltonian. Tethered FiP35 WW domains with n = 2 and 3 domains are just slightly less stable than the monomer. The n = 4 oligomer is yet less stable, its expression yield is much lower than the monomer's, and depends on the purification tag used. The n = 5 plasmid did not express at all, indicating the sudden onset of aggregation past n = 4. Thus, tethered FiP35 has a critical nucleus size for inter-domain aggregation of n ≈ 4. According to our simulations, misfolded structures become increasingly prevalent as one proceeds from monomer to pentamer, with extended inter-domain beta sheets appearing first, then multi-sheet 'intramolecular amyloid' structures, and finally novel motifs containing alpha helices. We discuss the implications of our results for oligomeric aggregate formation and structure, transient aggregation of proteins whilst folding, as well as for protein evolution that starts with repeat proteins.


Cell surface-tethered IL-12 repolarizes the tumor immune microenvironment to enhance the efficacy of adoptive T cell therapy.
Immune-activating cytokines such as interleukin-12 (IL-12) hold strong potential for cancer immunotherapy but have been limited by high systemic toxicities. We describe here an approach to safely harness cytokine biology for adoptive cell therapy through uniform and dose-controlled tethering onto the surface of the adoptively transferred cells. Tumor-specific T cells tethered with IL-12 showed superior antitumor efficacy across multiple cell therapy models compared to conventional systemic IL-12 coadministration. Mechanistically, the IL-12-tethered T cells supported a strong safety profile by driving interferon-γ production and adoptively transferred T cell activity preferentially in the tumor. Immune profiling revealed that the tethered IL-12 reshaped the suppressive tumor immune microenvironment, including triggering a pronounced repolarization of monocytic myeloid-derived suppressor cells into activated, inflammatory effector cells that further supported antitumor activity. This tethering approach thus holds strong promise for harnessing and directing potent immunomodulatory cytokines for cell therapies while limiting systemic toxicities.


α-Ketoglutarate attenuates Wnt signaling and drives differentiation in colorectal cancer.
Genetic-driven deregulation of the Wnt pathway is crucial but not sufficient for colorectal cancer (CRC) tumourigenesis. Here, we show that environmental glutamine restriction further augments Wnt signaling in APC mutant intestinal organoids to promote stemness and leads to adenocarcinoma formation in vivo via decreasing intracellular alpha-ketoglutarate (aKG) levels. aKG supplementation is sufficient to rescue low-glutamine induced stemness and Wnt hyperactivation. Mechanistically, we found that aKG promotes hypomethylation of DNA and histone H3K4me3, leading to an upregulation of differentiation-associated genes and downregulation of Wnt target genes, respectively. Using CRC patient-derived organoids and several in vivo CRC tumour models, we show that aKG supplementation suppresses Wnt signaling and promotes cellular differentiation, thereby significantly restricting tumour growth and extending survival. Together, our results reveal how metabolic microenvironment impacts Wnt signaling and identify aKG as a potent antineoplastic metabolite for potential differentiation therapy for CRC patients.


Preconception Phthalate Exposure and Women's Reproductive Health: Pregnancy, Pregnancy Loss, and Underlying Mechanisms.
Phthalates are endocrine-disrupting chemicals linked to adverse pregnancy outcomes. Despite the sensitivity of female reproductive processes to oxidation-reduction reaction stress and endocrine disruption, evidence for the impact of women's phthalate exposure on the ability to establish and maintain pregnancy has been inconclusive. We aimed to determine the relationship of preconception phthalate metabolite exposure with a) fecundability and pregnancy loss and b) markers of potential biological mechanisms, including reproductive hormones, inflammation, and oxidative stress. Data were collected from the Effects of Aspirin in Gestation and Reproduction (EAGeR) trial, a preconception study following 1,228 women who were attempting pregnancy, for up to six menstrual cycles and throughout pregnancy if they became pregnant. Twenty phthalate metabolites were measured in a consecutive 3-d pooled urine sample at enrollment. Pregnancy was determined through urinary human chorionic gonadotropin (hCG) at the expected date of menses during each cycle and pregnancy loss as an observed loss following positive hCG. Highly sensitive C-reactive protein (hsCRP) and isoprostanes were measured at enrollment, and reproductive hormones were measured during the follicular phase, ovulation, and luteal phase. Discrete-time Cox proportional hazards models evaluated the relationship of phthalate metabolites with fecundability and weighted Poisson models with robust variance evaluated the risk of pregnancy loss. An interquartile range (IQR) higher mono-(2-ethylhexyl) phthalate [fecundability odds ratio (FOR)=0.88; 95% confidence interval (CI): 0.78, 1.00], mono-butyl phthalate (FOR=0.82; 95% CI: 0.70, 0.96), and mono-benzyl phthalate (FOR=0.85; 95% CI: 0.74, 0.98) was associated with lower fecundability. No consistent associations were observed with pregnancy loss. Preconception phthalates were consistently associated with higher hsCRP and isoprostanes, as well as lower estradiol and higher follicle-stimulating hormone across the menstrual cycle. Women's preconception exposure to phthalates was associated with lower fecundability, changes in reproductive hormones, and increased inflammation and oxidative stress. The pre- and periconception periods may represent sensitive windows for intervening to limit the reproductive toxicity of phthalate exposure. https://doi.org/10.1289/EHP12287.


Non-invasive ventilation for very old patients with limitations to respiratory care in half-open geriatric ward: experience on a consecutive cohort of patients.
A leading role for non-invasive ventilation (NIV), as comfort treatment or palliative care, is actually recognized for very old patients suffering from ARF. NIV was frequently used in both ICU and respiratory ICU (RICUs) for very old patients and it is associated with a reduced rate of endotracheal intubations and mortality. This study aims to evaluate the effects of NIV, performed in a setting of half-open geriatric ward with family support, in a cohort of very old patients with ARF and DNI decision. A consecutive cohort of 20 very old patients with DNI decision was admitted in our 26-bed geriatric ward during a 6 months' period. DNI decision was obtained in emergency room with an intensive care physician supported by a psychologist. Pressure support ventilation was the first choice of NIV. NIV has been performed by three adequately trained geriatricians, with one of them experienced in ICU, and in close collaboration with intensive care physicians. Arterial blood gases, to assess the response to ventilation, were obtained after 1, 6 and 12 h. NIV settings were modified according to arterial blood gas analyses or respiratory fatigue, if needed. Therefore, 75% of patients were discharged home and 12 out of 20 patients had home respiratory support. PaO2/FiO2 ratio and pH increased while PaCO2 decreased during the 12 h of NIV with statistical significance. At the admission, alive patients had PaCO2 significantly lower than dead patients. After 12 h, alive patients had a better pH than dead patients. Dead patients experienced more complication than survivors. Very old DNI patients with ARF could be treated with NIV in half-open geriatric ward with trained physicians and nurses. The presence of family members may improve patients' comfort and reduce anxiety level even at the end of life. Further studies are needed to address the effective role of NIV in very old patients with DNI decisions.


Passive concentration dynamics incorporated into the library IB2d, a two-dimensional implementation of the immersed boundary method.
In this paper, we present an open-source software library that can be used to numerically simulate the advection and diffusion of a chemical concentration or heat density in a viscous fluid where a moving, elastic boundary drives the fluid and acts as a source or sink. The fully-coupled fluid-structure interaction problem of an elastic boundary in a viscous fluid is solved using Peskin's immersed boundary method. The addition or removal of the concentration or heat density from the boundary is solved using an immersed boundary-like approach in which the concentration is spread from the immersed boundary to the fluid using a regularized delta function. The concentration or density over time is then described by the advection-diffusion equation and numerically solved. This functionality has been added to our software library,IB2d, which provides an easy-to-use immersed boundary method in two dimensions with full implementations in MATLAB and Python. We provide four examples that illustrate the usefulness of the method. A simple rubber band that resists stretching and absorbs and releases a chemical concentration is simulated as a first example. Complete convergence results are presented for this benchmark case. Three more biological examples are presented: (1) an oscillating row of cylinders, representative of an idealized appendage used for filter-feeding or sniffing, (2) an oscillating plate in a background flow is considered to study the case of heat dissipation in a vibrating leaf, and (3) a simplified model of a pulsing soft coral where carbon dioxide is taken up and oxygen is released as a byproduct from the moving tentacles. This method is applicable to a broad range of problems in the life sciences, including chemical sensing by antennae, heat dissipation in plants and other structures, the advection-diffusion of morphogens during development, filter-feeding by marine organisms, and the release of waste products from organisms in flows.


Genome stability: What we have learned from cohesinopathies.
Cohesin is a multiprotein complex involved in many DNA-related processes such as proper chromosome segregation, replication, transcription, and repair. Mutations in cohesin gene pathways are responsible for human diseases, collectively referred to as cohesinopathies. In addition, both cohesin gene expression dysregulation and mutations have been identified in cancer. Cohesinopathy cells are characterized by genome instability (GIN) visualized by a constellation of markers such as chromosome aneuploidies, chromosome aberrations, precocious sister chromatid separation, premature centromere separation, micronuclei formation, and sensitivity to genotoxic drugs. The emerging picture suggests that GIN observed in cohesinopathies may result from the synergistic effects of the multiple cohesin dysfunctions. © 2016 Wiley Periodicals, Inc.


Comprehensive Computer-Aided Decision Support Framework to Diagnose Tuberculosis From Chest X-Ray Images: Data Mining Study.
Tuberculosis (TB) is one of the most infectious diseases that can be fatal. Its early diagnosis and treatment can significantly reduce the mortality rate. In the literature, several computer-aided diagnosis (CAD) tools have been proposed for the efficient diagnosis of TB from chest radiograph (CXR) images. However, the majority of previous studies adopted conventional handcrafted feature-based algorithms. In addition, some recent CAD tools utilized the strength of deep learning methods to further enhance diagnostic performance. Nevertheless, all these existing methods can only classify a given CXR image into binary class (either TB positive or TB negative) without providing further descriptive information. The main objective of this study is to propose a comprehensive CAD framework for the effective diagnosis of TB by providing visual as well as descriptive information from the previous patients' database. To accomplish our objective, first we propose a fusion-based deep classification network for the CAD decision that exhibits promising performance over the various state-of-the-art methods. Furthermore, a multilevel similarity measure algorithm is devised based on multiscale information fusion to retrieve the best-matched cases from the previous database. The performance of the framework was evaluated based on 2 well-known CXR data sets made available by the US National Library of Medicine and the National Institutes of Health. Our classification model exhibited the best diagnostic performance (0.929, 0.937, 0.921, 0.928, and 0.965 for F1 score, average precision, average recall, accuracy, and area under the curve, respectively) and outperforms the performance of various state-of-the-art methods. This paper presents a comprehensive CAD framework to diagnose TB from CXR images by retrieving the relevant cases and their clinical observations from the previous patients' database. These retrieval results assist the radiologist in making an effective diagnostic decision related to the current medical condition of a patient. Moreover, the retrieval results can facilitate the radiologists in subjectively validating the CAD decision.


Design optimization of a fast-neutron detector with scintillating fibers for triton burnup experiments at fusion experimental devices.
Time-resolved triton burnup studies have been carried out to estimate the behavior of alpha particles in DD fusion experimental devices. In those studies, 14 MeV neutrons emitted through DT reactions in DD plasmas should be measured selectively in the backgrounds of DD neutrons and gamma rays. For this purpose, a scintillating-fiber (Sci-Fi) based fast-neutron detector has been adapted because of its advantages such as fast response, design flexibility in detection efficiency by changing the number of Sci-Fi, and discrimination property against 2.4 MeV neutrons produced through DD reactions and gamma rays. However, its length had conventionally been set to around 10 cm without an optimization study of its design parameters to meet the requirements as 14 MeV neutron detector. In the present study, we tested three types of Sci-Fi detectors with three different lengths and compared with the simulated results of energy deposition, through which we tried to understand the phenomena in the detection process of fast neutrons. From the results, it has been shown that, due to the self-shielding of neutrons by Sci-Fi and the attenuation of scintillation photons during the transmission process to the photomultiplier tube, the optimal length of Sci-Fi is concluded to be about 6 cm.


DRUDIT: web-based DRUgs DIscovery Tools to design small molecules as modulators of biological targets.
New in silico tools to predict biological affinities for input structures are presented. The tools are implemented in the DRUDIT (DRUgs DIscovery Tools) web service. The DRUDIT biological finder module is based on molecular descriptors that are calculated by the MOLDESTO (MOLecular DEScriptors TOol) software module developed by the same authors, which is able to calculate more than one thousand molecular descriptors. At this stage, DRUDIT includes 250 biological targets, but new external targets can be added. This feature extends the application scope of DRUDIT to several fields. Moreover, two more functions are implemented: the multi- and on/off-target tasks. These tools applied to input structures allow for predicting the polypharmacology and evaluating the collateral effects. The applications described in the article show that DRUDIT is able to predict a single biological target, to identify similarities among biological targets, and to discriminate different target isoforms. The main advantages of DRUDIT for the scientific community lie in its ease of use by worldwide scientists and the possibility to be used also without specific, and often expensive, hardware and software. In fact, it is fully accessible through the WWW from any device to perform calculations. Just a click or a tap can start tasks to predict biological properties for new compounds or repurpose drugs, lead compounds, or unsuccessful compounds. To date, DRUDIT is supported by four servers each able to execute 8 jobs simultaneously. The web service is accessible at the www.drudit.com URL and its use is free of charge. Supplementary data are available at Bioinformatics online.


Strategies based on metal-based nanoparticles for hypoxic-tumor radiotherapy.
Radiotherapy (RT) is one of the most effective and frequent clinical cancer treatments. Nevertheless, RT can cause damage to normal tissues around tumors under high-dose ionizing radiation. Inspired by versatile metal-based nanomaterials, great efforts have been devoted to developing nanomaterials with high-Z metal elements as radiosensitizers by depositing more energy into tumors for RT enhancement. However, these metal-based nanomaterial-mediated RTs are highly O2-dependent. Unfortunately, O2 concentrations within the majority of solid tumors exhibit low levels, which seriously hampers the antitumor efficacy of these nanomaterials during RT. Therefore, the development of novel metal-based nanomaterials as radiosensitizers capable of avoiding the radioresistance induced by tumor hypoxia is highly desirable and important. Currently, the most effective approaches to reverse the radioresistance of hypoxic tumors are to introduce nanomaterials with O2-elevating ability by delivering exogenous O2, generating O2 in situ, increasing intratumoral blood flow, or reducing HIF-1 expression to harness the O2 level in solid tumors. Besides these, recently, some innovative and simple strategies by employing nanoradiosensitizers with diminished oxygen dependence have also been applied to combat unmet hypoxic challenges, in which nanoradiosensitizers can target tumor hypoxia for selective RT, enhance oxygen-independent ROS generation, or combine with non-oxygen dependent cancer therapies for synergistic treatments. These approaches and strategies provide new avenues for enhanced hypoxic-tumor RT. Nevertheless, an overall review aiming specifically at these strategies is still rare. Herein, we present an overview about recent advances in metal-based nanomaterials for hypoxic-tumor RT, and give a detailed discussion about the design and working mechanisms of these strategies in their application of RT. Finally, current challenges and future perspectives are also pointed out in this field.


Modulation of expression and activity of cytochrome P450s and alteration of praziquantel kinetics during murine schistosomiasis.
In this study, we investigated the expression and activity of liver cytochrome P450s (CYPs) and praziquantel (PZQ) kinetics in mice infected with Schistosoma mansoni. Swiss Webster (SW) mice of both genders were infected (100 cercariae) on postnatal day 10 and killed on post-infection days (PIDs) 30 or 55. Non-infected mice of the same age and sex served as controls. Regardless of mouse sex, infection depressed the activities of CYP1A [ethoxy/methoxy-resorufin-O-dealkylases (EROD/MROD)], 2B9/10 [pentoxy/benzyloxy-resorufin-O-dealkylases (PROD, BROD)], 2E1 [p-nitrophenol-hydroxylase (PNPH)] and 3A11 [erythromycin N-demethylase (END)] on PID 55 but not on PID 30. On PID 55, infection decreased liver CYP mRNA levels (real-time reverse transcription-polymerase chain reaction). On PID 30, whereas mRNA levels remained unaltered in males, they were depressed in females. Plasma PZQ (200 and 400 mg/kg body weight intraperitoneally) levels were measured (high-performance liquid chromatography) at different post-treatment intervals. In males and females, infection delayed the PZQ clearance on PID 55, but not on PID 30. Therefore, it can be concluded that schistosomiasis down-modulated CYP expression and activity and delayed PZQ clearance on PID 55, when a great number of parasite eggs were lodged in the liver. On PID 30, when egg-laying was initiated by the worms, no change of CYP expression and activity was found, except for a depression of CYP1A2 and 3A11 mRNAs in female mice.


Novel mutation that causes a structural change in a lipoprotein in the outer membrane of Escherichia coli.
A novel mutation which caused a structural change in a lipoprotein in the outer-membrane has been found in Escherichia coli K-12. The lipoprotein of the wild-type strain is known to have a peculiar amino terminal structure: glycerylcysteine with two fatty acids attached by ester linkages and one fatty acid by an amide linkage. In contrast to the wild-type lipoprotein, the mutant lipoproteins is isolated from the E. coli envelope as a dimer of molecular weight of about 15,000. The dimer can be reduced by mercaptoethanol to the lipoprotein monomer of molecular weight of about 7,500. The monomer has a free thiol group which is susceptible to monoiodacetie mutant lipoprotein is extremely low in comparison with that into the wild-type lipoprotein. These results suggest that the mutant is defective in transferring a glycerol group to the thiol group of the amino terminal cysteine residue of the lipoprotein. The gene responsible for this modification reaction has been located at 36.5 min on the E. coli chromosome.


The tubulin database: Linking mutations, modifications, ligands and local interactions.
Microtubules are polymeric filaments, constructed of α-β tubulin heterodimers that underlie critical subcellular structures in eukaryotic organisms. Four homologous proteins (γ-, δ-, ε- and ζ-tubulin) additionally contribute to specialized microtubule functions. Although there is an immense volume of publicly available data pertaining to tubulins, it is difficult to assimilate all potentially relevant information across diverse organisms, isotypes, and categories of data. We previously assembled an extensive web-based catalogue of published missense mutations to tubulins with >1,500 entries that each document a specific substitution to a discrete tubulin, the species where the mutation was described and the associated phenotype with hyperlinks to the amino acid sequence and citation(s) for research. This report describes a significant update and expansion of our online resource (TubulinDB.bio.uci.edu) to nearly 18,000 entries. It now encompasses a cross-referenced catalog of post-translational modifications (PTMs) to tubulin drawn from public datasets, primary literature, and predictive algorithms. In addition, tubulin protein structures were used to define local interactions with bound ligands (GTP, GDP and diverse microtubule-targeting agents) and amino acids at the intradimer interface, within the microtubule lattice and with associated proteins. To effectively cross-reference these datasets, we established a universal tubulin numbering system to map entries into a common framework that accommodates specific insertions and deletions to tubulins. Indexing and cross-referencing permitted us to discern previously unappreciated patterns. We describe previously unlinked observations of loss of PTM sites in the context of cancer cells and tubulinopathies. Similarly, we expanded the set of clinical substitutions that may compromise MAP or microtubule-motor interactions by collecting tubulin missense mutations that alter amino acids at the interface with dynein and doublecortin. By expanding the database as a curated resource, we hope to relate model organism data to clinical findings of pathogenic tubulin variants. Ultimately, we aim to aid researchers in hypothesis generation and design of studies to dissect tubulin function.


Back-Projection Cortical Potential Imaging: Theory and Results.
Electroencephalography (EEG) is the single brain monitoring technique that is non-invasive, portable, passive, exhibits high-temporal resolution, and gives a directmeasurement of the scalp electrical potential. Amajor disadvantage of the EEG is its low-spatial resolution, which is the result of the low-conductive skull that "smears" the currents coming from within the brain. Recording brain activity with both high temporal and spatial resolution is crucial for the localization of confined brain activations and the study of brainmechanismfunctionality, whichis then followed by diagnosis of brain-related diseases. In this paper, a new cortical potential imaging (CPI) method is presented. The new method gives an estimation of the electrical activity on the cortex surface and thus removes the "smearing effect" caused by the skull. The scalp potentials are back-projected CPI (BP-CPI) onto the cortex surface by building a well-posed problem to the Laplace equation that is solved by means of the finite elements method on a realistic head model. A unique solution to the CPI problem is obtained by introducing a cortical normal current estimation technique. The technique is based on the same mechanism used in the well-known surface Laplacian calculation, followed by a scalp-cortex back-projection routine. The BP-CPI passed four stages of validation, including validation on spherical and realistic head models, probabilistic analysis (Monte Carlo simulation), and noise sensitivity tests. In addition, the BP-CPI was compared with the minimum norm estimate CPI approach and found superior for multi-source cortical potential distributions with very good estimation results (CC >0.97) on a realistic head model in the regions of interest, for two representative cases. The BP-CPI can be easily incorporated in different monitoring tools and help researchers by maintaining an accurate estimation for the cortical potential of ongoing or event-related potentials in order to have better neurological inferences from the EEG.


[Room tilt illusion: Report of two cases and terminological review].
The room tilt illusion is a transient misperception of the visual image as tilted on its side or even upside down; in this case it has been termed acute upside down reversal of vision. We report on two cases of room tilt illusion as manifestation of VIII nerve neuritis (herpes-zoster infection) and cerebellar hemorrhage. Room tilt illusion has been reported in association with vertebrobasilar stroke, migraine, multiple sclerosis, epilepsy and labyrinthine disorders. The pathophysiology of this rare visual illusion has been related to a lesion of the visual or vestibulo-otolith pathways. In animals the neurones of the parieto-insular vestibular cortex areas are multisensory. So, they can respond to somatosensory, optokinetic and visual stimuli. In humans the knowledge about vestibular cortex function and localization is less precise than in animals. However, we propose a disorder of multisensorial vestibular cortex, resulting from a lession of vestibular pathways or association cortex, as mechanism of this phenomenon.


The potential of transformative video design for improving caregiver's wellbeing.
The existing interventions for informal caregivers assist with managing health outcomes of the role burden. However, the deeper meaning-making needs of informal caregivers have been generally neglected. This paper reflects on the meaning-making needs of informal caregivers, through the theory of narrative identity, and proposes a new approach - the Transformative Video Design technique delivered via video storytelling. Transformative Video Design assists informal caregivers to re-create a cohesive caregiving story and incorporate it into the narrative identity. The technique is used as a stimulus for triggering the self-re-structure within the narrative identity and facilitating role transformation.


Contrasting effects of sampling scale on insect herbivores distribution in response to canopy structure.
Species diversity of insect herbivores associated to canopy may vary local and geographically responding to distinct factors at different spatial scales. The aim of this study was to investigate how forest canopy structure affects insect herbivore species richness and abundance depending on feeding guilds' specificities. We tested the hypothesis that habitat structure affects insect herbivore species richness and abundance differently to sap-sucking and chewing herbivore guilds. Two spatial scales were evaluated: inside tree crowns (fine spatial cale) and canopy regions (coarse spatial scale). In three sampling sites we measured 120 tree crowns, grouped n five points with four contiguous tree crowns. Insects were sampled by beating method from each crown and data were summed up for analyzing each canopy region. In crowns (fine spatial scale) we measured habitat tructure: trunk circumference, tree height, canopy depth, number of ramifications and maximum ramification level. In each point, defined as a canopy region (coarse spatial scale), we measured habitat structure using a vertical cylindrical transect: tree species richness, leaf area, sum of strata heights and maximum canopy height. A principal component analysis based on the measured variables for each spatial scale was run to estimate habitat structure parameters. To test the effects of habitat structure upon herbivores, different general linear models were adjusted using the first two principal components as explanatory variables. Sap-sucking insect species richness and all herbivore abundances increased with size of crown at fine spatial scale. On the other hand, chewer species richness and abundance increased with resource quantity at coarse scale. Feeding specialization, resources availability, and agility are discussed as ecological causes of the found pattern.


In vitro release, rheological, and stability studies of mefenamic acid coprecipitates in topical formulations.
A suitable topical formulation of mefenamic acid was developed in order to eliminate the gastrointestinal disorders associated with its oral administration. Drug coprecipitates prepared with different polymers at various drug-to-polymer ratios improved drug solubility and dissolution compared to pure drug and physical mixtures. PVP polymers (ratio 1:4) produced the best results. Aqueous ionic cream, ointments of absorption and water soluble bases and gels of methylcellulose, carboxymethylcellulose sodium, HPMC, Carbopol® 934 and 940, and Pluronic® F127 bases containing 1-10% drug as coprecipitates of PVP polymers (1:4) were prepared. The highest drug release was achieved at 1% drug concentration from water soluble base and methylcellulose among cream/ointment and gel bases, respectively. Gels, in general yielded better release than creams/ointments. All tested medicated creams/ointments exhibited plastic flow while all gels conformed to pseudoplasticity. Most of them showed thixotropy, a desired property of topical preparations. Stability studies revealed that HPMC and methylcellulose had the smallest changes in drug content, viscosity, and pH among the formulations. Considering drug release, rheological properties, and stability, methylcellulose gel containing 1% drug as coprecipitates of PVP K90 was the best among the studied formulations, was promising for improving bioavailability of mefenamic acid and can be used in future studies.


Appropriateness in Dentistry: A Survey Discovers Improper Procedures in Oral Medicine and Surgery.
The aim of this study was to assess appropriateness of diagnostic exams, treatments, and procedures among Italian dental practitioners. A questionnaire with multiple responses on topics of dentistry and oral medicine was administered to a sample of 198 Italian dental practitioners. Information on characteristics of the respondents was also collected. Descriptive statistics and multiple correspondence analysis (MCA) were applied. Data were analyzed using R software (version 3.3.2). The survey respondents included Doctors of Medicine (MD) (54/198 = 27%) with or without specialty in dentistry (33% versus 67%), Doctors of Dental Surgery (DDS) (144/198 = 73%), DDS with specialty in orthodontics (7%), and DDS with specialty in oral surgery (4%). Mandatory procedures in dental and oral medicine education and training include (a) prescription of antibiotics before/after oral surgery procedures; (b) prevention strategies for oral cancer, and (c) prescription of dental X-ray examinations (41%, 52%, and 48%, resp.). On examining the results of the survey, it is evident that information and implementation of the above mentioned procedures are crucially needed. Our results confirm the necessity to reduce inappropriate practices in dentistry, implementing formation and information, leading to correct prescriptions, and optimizing patients' oral health. This coincides with the Italian Slow Medicine program entitled "Fare di più non significa fare meglio-Choosing Wisely Italy," which has also motivated this study.


Amoxicillin-induced aseptic meningoencephalitis.
Meningitis is usually produced by an infectious agent, but there are multiple noninfectious causes. Drug-induced aseptic meningitis (DIAM) is an important entity and has been reported as an uncommon adverse reaction with numerous agents. Thus, DIAM constitutes a diagnostic and patient management challenge. We present a patient with three episodes of aseptic meningitis due to amoxicillin, and then review the literature on this rare idiosyncratic event which may occur after local or systemic drug administration. A 77-year-old man was admitted to our hospital with fever, headache, and neck stiffness. Seven days before admission he had a dental and gingival inflammation. He was treated with two oral doses of 500 mg daily of amoxicillin for one week. The seventh day he awoke with the complaints that prompted hospital admittance. Amoxicillin was stopped 1 day before his admission. From his history we knew of two similar episodes: The first episode was after a dental procedure 3 months before this incident. He had received a 1-week course of postprocedure amoxicillin of 500 mg daily and had similar headache, fever, and chills during the entire course of treatment. He wasn't admitted to the hospital, because he stopped taking amoxicillin and he felt spontaneous pain relief after taking symptomatic pain treatment. The second episodes was 6 months after his first admission, he had been admitted to our hospital with the same symptoms. Amoxicillin was stopped and changed with intravenous (IV) ceftriaxone (CTRX) for 10 days due to suspected partial untreated meningitis. The patient improved rapidly within 2 days and was discharged from the hospital. On the basis of these three confirmed episodes of meningitis after recurrent exposure to amoxicillin, with repetitive negative testing for viral, bacterial, and mycobacterial micro-organisms, we diagnosed aseptic meningitis induced by amoxicillin. To our knowledge, this is the seventh well documented publication of such a severe side effect of a commonly used antibiotic.


Deep-Learning-Based Segmentation of Small Extracellular Vesicles in Transmission Electron Microscopy Images.
Small extracellular vesicles (sEVs) are cell-derived vesicles of nanoscale size (~30-200 nm) that function as conveyors of information between cells, reflecting the cell of their origin and its physiological condition in their content. Valuable information on the shape and even on the composition of individual sEVs can be recorded using transmission electron microscopy (TEM). Unfortunately, sample preparation for TEM image acquisition is a complex procedure, which often leads to noisy images and renders automatic quantification of sEVs an extremely difficult task. We present a completely deep-learning-based pipeline for the segmentation of sEVs in TEM images. Our method applies a residual convolutional neural network to obtain fine masks and use the Radon transform for splitting clustered sEVs. Using three manually annotated datasets that cover a natural variability typical for sEV studies, we show that the proposed method outperforms two different state-of-the-art approaches in terms of detection and segmentation performance. Furthermore, the diameter and roundness of the segmented vesicles are estimated with an error of less than 10%, which supports the high potential of our method in biological applications.


Estimating human indoor exposure to elemental mercury from broken compact fluorescent lamps (CFLs).
The 2008 EU regulation, which prohibits conventional incandescent light bulbs, is to be implemented in phases, completing in 2012. One of the possible substitutes is the compact fluorescent lamp (CFL), which, however, does contain up to 5 mg of mercury in its elemental or amalgamated form. The question arises as to the possible exposure of individuals to mercury as a result of lamp breakage during operation or when disconnected from the power supply. Therefore, an apparatus was built to shatter CFLs and drop the shards onto glycol-modified polyethylene terephthalate, a carpeted floor, or laminate floor under defined climatic parameters and operating conditions. Six CFLs of different types and mercury content were studied. After the breakage of a common CFL containing liquid mercury, concentrations up to 8000 ng/m(3) were reached in the chamber. Much lower peak values were obtained with amalgam-type lamps (414 ng/m(3)) or with lamps with a shatter-proof coating (60 ng/m(3)). It was found that ventilation can considerably reduce the indoor air concentration within 20 min. Acute health effects would only be expected if the mercury is not removed immediately. Careful collection and disposal of the lamp fragments would also prevent dwellers from the risk of long-term exposure. After accidental breakage of a compact fluorescent lamp (CFL) indoors, dwellers could be exposed to high mercury concentrations. From the results of our studies in test chambers and real rooms using different lamp types and scenarios, it was possible to estimate the possible human uptake of mercury by inhalation. Immediate action is important to reduce indoor mercury concentrations to a minimum level. The first step is to maximize ventilation followed by careful collection of spilled mercury.


TRPV3 and Itch: The Role of TRPV3 in Chronic Pruritus according to Clinical and Experimental Evidence.
Itching is a sensory phenomenon characterized by an unpleasant sensation that makes you want to scratch the skin, and chronic itching diminishes the quality of life. In recent studies, multiple transient receptor potential (TRP) channels present in keratinocytes or nerve endings have been shown to engage in the propagation of itch signals in chronic dermatological or pruritic conditions, such as atopic dermatitis (AD) and psoriasis (PS). TRPV3, a member of the TRP family, is highly expressed in the epidermal keratinocytes. Normal TRPV3 signaling is essential for maintaining epidermal barrier homeostasis. In recent decades, many studies have suggested that TRPV3 contributes to detecting pruritus signals. Gain-of-function mutations in TRPV3 in mice and humans are characterized by severe itching, hyperkeratosis, and elevated total IgE levels. These studies suggest that TRPV3 is an important channel for skin itching. Preclinical studies have provided evidence to support the development of TRPV3 antagonists for treating inflammatory skin conditions, itchiness, and pain. This review explores the role of TRPV3 in chronic pruritus, collating clinical and experimental evidence. We also discuss underlying cellular and molecular mechanisms and explore the potential of TRPV3 antagonists as therapeutic agents.


Sample collection and platelet function testing: influence of vacuum or aspiration principle on PFA-100 test results.
As for other tests of hemostasis, the investigation of platelet function is highly vulnerable to a broad series of preanalytical variables, which span from patient preparation to the final analysis of the specimen and issuance of test results. In particular, there remains much controversy about the influence of manual or vacuum aspiration of blood into primary collection tubes on platelet function testing. Accordingly, we investigated this for the PFA-100. In 12 healthy volunteers, a sample labeled as 'BD-V' was drawn into a 2.7 ml BD Vacutainer tube, whereas two additional samples were collected from the opposite arm into 5.0 ml Sarstedt S-Monovette tubes by vacuum (SD-V) or manual aspiration (SD-A). All sample were tested on PFA-100 with collagen and ADP (CADP) or collagen and epinephrine (CEPI). The values of both CEPI and CADP obtained in SD-A samples were significantly lower than those obtained in SD-V and BD-V tubes, whereas those of the two evacuated tubes did not significantly differ. On average, CEPI values were prolonged by 11% in SD-V and 13% in BD-V, whereas those of CADP were prolonged by 14% in SD-V and 10% in BD-V, respectively. These findings suggests that the lower shear stress generated by the manual aspiration of blood into the primary collection tube would prevent spurious hyper-activation of platelets, thus, preserving the integrity of their function for subsequent testing on PFA-100. This study underscores the need to define or validate local reference ranges for the PFA-100 based on the collection tube used. Different reference ranges of both CEPI and CADP may also be advisable when venous blood samples are collected with manual aspiration or vacuum principle.


[Recurrences of supratentorial meningiomas. Apropos of 17 cases].
17 cases of recurrent meningiomas observed during a 10 years period are reviewed. Meningiomas with a second localisation different from the first were excluded from the study. The tumor recurrence occurred mostly during the first five years after the surgical procedure. One case with femoral and pulmonary metastasis is described. Several factors may play a role in the genesis of the recurrence: age, sex, the tumor localisation, CT scan, the importance of the tumor ablation, the histology. In almost all cases a radiotherapy was carried out after the second surgical procedure. This retrospective study suggests the need for multifactorial prospective analysis with clinical, paraclinical and fundamental factors. Such a study would be useful to precise the natural history of meningiomas.


Clinical significance of antiphospholipid protein antibodies. Receiver operating characteristics plot analysis.
To investigate the utility of receiver operating characteristic (ROC) analysis in determining the strength of association between various antiphospholipid and anti-protein cofactor antibodies (aPA) and thrombosis, pregnancy morbidity, and thrombocytopenia. Clinical and laboratory variables were retrospectively studied in 204 patients: 160 with systemic lupus erythematosus (SLE), 22 with lupus-like syndrome (SLE-LS), and 22 with primary antiphospholipid syndrome (APS). Laboratory evaluation included detection of lupus anticoagulant (LAC) and measurement of IgG and IgM anticardiolipin (aCL), antiphosphatidylserine (aPS), antiphosphatidylinositol (aPI), anti-beta 2 glycoprotein I (a beta 2GPI), and antiprothrombin (aPT) antibodies. ROC plot analysis was used to determine the clinical accuracy of aPA tests, and calculate cut-off values which best associate with clinical symptoms typical for APS. The LAC was associated with a history of thrombosis [odds ratio (OR): 3.04; 95% confidence interval (CI): 1.5-6.2] and even more strongly with recurrent fetal loss (OR: 8.7; 95%CI: 2.8-26.7). ROC plot analysis revealed that the most accurate test for thrombosis was aCL IgG (ROC-derived cutoff value > 17.2 GPL; OR: 3.69; 95% CI: 1.8-7.4), for recurrent fetal loss, aPI IgG [> 22.1 theoretical units (TU); OR: 6.21; 95%CI: 2.1-18.5], closely followed by aCL IgG and a beta 2GPI IgG, and for thrombocytopenia aPS IgM (> 6.7 TU; OR: 1.9; 95%CI: 1.04-3.4). Among 182 autoimmune patients (SLE + SLE-LS), 6.6% presented clinical symptoms of APS without classic aPA (LAC and/or aCL), but with elevated levels of antibodies against other phospholipids, mainly aPI IgM. A laboratory that evaluates APS patients should establish its own threshold values for aPA tests. We suggest that ROC plot analysis is a valuable tool in establishing cutoff values. LAC and aCL determinations seem sufficient for the majority of laboratories. However, in specialized centers other tests should be available to detect those patients with clinical symptoms for APS but who are positive for antiphospholipid antibodies other than aCL and the LAC.


Functional design of horse hoof keratin: the modulation of mechanical properties through hydration effects.
Tensile moduli and J-integral fracture toughness values were determined for horse hoof-wall keratin at four hydration levels. The stiffness of hoof-wall was influenced by water content to a greater degree than is the stiffness of other mammalian hard keratins. Young's modulus increased from 410 MPa at 100% relative hydration (RH) to 14.6 GPa at 0% RH. Fracture toughness was maximal (22.8 kJ m-2) at an intermediate hydration (75% RH), which represents a two-fold increase over both fully hydrated and dehydrated material. Maximum fracture toughness occurred at a hydration level which is within the range that has been found in vivo in the hoof wall. These results lead to the hypothesis that the density of secondary bonding sites within the hoof-wall keratin matrix proteins provides the hoof organ with the means to modulate tissue properties, even though this epidermal tissue functions after the cells have died.


CTLA-4 dysregulation of self/tumor-reactive CD8+ T-cell function is CD4+ T-cell dependent.
Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) maintains peripheral tolerance by suppressing T-cell activation and proliferation but its precise role in vivo remains unclear. We sought to elucidate the impact of CTLA-4 expression on self/tumor-reactive CD8(+) T cells by using the glycoprotein (gp) 100-specific T-cell receptor (TCR) transgenic mouse, pmel-1. pmel-1 CLTA-4(-/-) mice developed profound, accelerated autoimmune vitiligo. This enhanced autoimmunity was associated with a small but highly activated CD8(+) T-cell population and large numbers of CD4(+) T cells not expressing the transgenic TCR. Adoptive transfer of pmel-1 CLTA-4(-/-) CD8(+) T cells did not mediate superior antitumor immunity in the settings of either large established tumors or tumor challenge, suggesting that the mere absence of CTLA-4-mediated inhibition on CD8(+) T cells did not directly promote enhancement of their effector functions. Removal of CD4(+) T cells by crossing the pmel-1 CLTA-4(-/-) mouse onto a Rag-1(-/-) background resulted in the complete abrogation of CD8(+) T-cell activation and autoimmune manifestations. The effects of CD4(+) CLTA-4(-/-) T cells were dependent on the absence of CTLA-4 on CD8(+) T cells. These results indicated that CD8(+) CLTA-4(-/-) T-cell-mediated autoimmunity and tumor immunity required CD4(+) T cells in which the function was dysregulated by the absence of CTLA-4-mediated negative costimulation.


The cytopathological spectrum of lymphomas in effusions in a tertiary center in Taiwan.
Lymphomas presenting in effusions could either be primary or secondary, with very limited data from Taiwan. We retrospectively reviewed effusion lymphomas from our archives in a tertiary center from July 2011 to June 2019. We identified 59 specimens from 43 patients, including 7 cases with primary effusion lymphoma (PEL) and 36, secondary effusion involvement. Half of the secondary cases presented concurrently with effusion lymphoma, while the remaining half-experienced effusion lymphoma during disease progression. All patients with PELs were males with a median age of 77 and presented with massive pleural effusion. None was HIV-related. Two (29%) PEL cases were positive for human herpes virus 8 (HHV8). The only case with plasmablastic phenotype in the PEL group was positive for both HHV8 and EBV. Four patients died shortly after diagnosis; while the remaining three were alive at the last follow-up (two at 13 months and one at 99 months). Of the secondary cases, diffuse large B-cell lymphoma/high grade B-cell lymphoma was the most common (n = 16, 44%), followed by mantle cell lymphoma (n = 5, 14%). Only 8 cases (22%) were T-cell neoplasms. Prognosis for patients with secondary effusion involvement was dismal, with 1- and 2-year overall survival rates at 17% and 8%, respectively. We found a wide cytopathological spectrum of effusion lymphoma in Taiwan. Most of our PEL cases were distinct from that defined in the World Health Organization scheme by a B-cell phenotype, HHV8-negativity, and absence of immunodeficiency. As compared to PEL cases, the prognosis of those with secondary involvement was extremely poor.


Effects of certain nucleoside analogues on human cytomegalovirus replication in vitro.
Four nucleoside analogues, 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-methyluracil (FMAU), -5-iodouracil (FIAU), -5-methylcytosine (FMAC) and -5-iodocytosine (FIAC), were studied for their effect on human cytomegalovirus (HCMV) replication in vitro. FMAU, FIAU, FMAC and FIAC showed antiviral activities for four strains of HCMV (Major, Clegg, D550 and Towne) in a plaque reduction assay, with a dose required for 50% inhibition (ED50) in the range of 0.1 to 0.65 microM. At a concentration of 1 microM-FMAU or -FIAC, the synthesis of five virus-specific late polypeptides of molecular weights 150 000, 120 000, 67000, 54000 and 27000 was entirely blocked. Quantification of Towne viral DNA synthesis, using complementary RNA-DNA hybridization with a Towne-specific cRNA probe, demonstrated a complete inhibition of HCMV DNA replication at 1 microM of FMAU or FIAC. After the removal of the inhibitors, however, viral DNA synthesis resumed, and infectious virus reappeared, indicating that the inhibition of HCMV replication by these nucleoside analogues was of a virostatic reversible type.