Distribution of neuropeptide immunoreactive nerve terminals within the subnuclei of the nucleus of the tractus solitarius of the rat.
The location of substance P, enkephalin and somatostatin (SRIF), and neurophysin II immunoreactive nerve terminals and preterminal processes in the caudal part of the nucleus of the tractus solitarius (nTS) was examined by the indirect immunofluorescence method for immunocytochemistry combined with cytoarchitectural identification of nuclear subgroups in the same tissue. In 22 Sprague-Dawley rats we examined 14-micrometers-thick serial sections of the dorsal medulla at levels from 1 mm caudal to 2 mm rostral to the obex. These sections were incubated with substance P, enkephalin, somatostatin, and neurophysin II antisera. All four peptides were examined in each case and five typical levels (two caudal and three rostral to the obex) were selected for comparison of terminal distribution between peptides. All sections were photographed under the fluorescence microscope and then counter-stained with cresyl violet. This method of analysis revealed distinct patterns of neuropeptide immunoreactivity in the subnuclei of the nTS that varied according to the level of the section. The nTS is responsible for integrating respiratory, cardiovascular (baroreceptor and cardiac), and gastrointestinal functions. The ventrolateral subnucleus (Vl)nTS, ventral subnucleus (v)nTS, interstitial subnucleus (ni)nTS, and intermediate subnucleus (nI)nTS are the major respiratory subnuclei with vlnTS and vnTS prominently associated with pulmonary afferents, ni associated with laryngeal afferents, and nI with tracheal afferents. The vlnTS, vnTS, and ni showed a moderate density of somatostatin-positive nerve terminals, scattered substance P and enkephalin immunoreactivity, and no neurophysin II-positive terminals. The nI showed moderate density of substance P immunoreactive nerve terminals. The subnuclei of the nTS receiving baroreceptor and chemoreceptor afferents--dorsolateral and dorsal (dl and d) subnuclei of nTS--showed scattered substance P immunoreactive nerve terminals. The commissural nucleus of nTS (ncom), which receives most of the cardiac afferents, showed a moderate density of enkephalin-positive immunoreactive nerve terminals. The medial subnucleus (m)nTS at levels rostral to the obex, the primary site for the termination of gastrointestinal afferents, showed substance P immunoreactivity in moderate amounts and weak immunoreactivity for all the other neuropeptides. An important result of these experiments was the observation that regions of the medulla adjacent to the nTS, i.e., the ventral parasolitarius region (vPSR), dorsal (d)PSR, and the periventricular region (PVR) showed the densest amounts of immunoreactive nerve terminals.(ABSTRACT TRUNCATED AT 400 WORDS)


Responses of phytoplankton to experimental fertilization with ammonium and phosphate in an African soda lake.
Phytoplankton abundance in tropical lakes is more often judged to be limited by nitrogen than phosphorus, but seldom does the evidence include controlled enrichments of natural populations. In January 1980 we performed the first experimental fertilization in an equatorial African soda lake, Lake Sonachi, a small, meromictic volcanic crater lake in Kenya. During our study the natural phytoplankton abundance was ca. 80 μg chl a/l, and the euphotic zone PO4 and NH4 concentrations were less than 0.5 μM. In the monimolimnion PO4 reached 180 μM and NH4 reached 4,600 μM. Replicate polyethylene cylinders (5 m long, 1.2 m3) were enriched to attain 10 μM PO4 and 100 μM NH4. Phytoplankton responses were measured as chlorophyll, cell counts and particulate N, P and C. After two days, the chlorophyll increase in the P treatment was significantly higher than the control (P<0.01) while the N treatment was not. After five days the molar N/P ratio of seston was the same in the N treatment and control (23) but only 6 in the P treatment. The molar N/P ratio of seston in an unenriched Lake Sonachi sample was 21 and in samples from Lakes Bogoria and Elmenteita, two shallow soda lakes in Kenya, the ratios were 12 and 70 respectively. We conclude that limitation of phytoplankton abundance by phosphorus can occur even in some tropical African soda lakes.


Investigations on purine and pyrimidine bases stacking associations in aqueous solutions by the fluorescence quenching method. I. Autoassociation of 2-aminopurine.
A general equation was derived, describing fluorescence quantum yield and lifetime of an autoassociating compound in liquid solutions. The autoassociation of 2-aminopurine in aqueous solution was examined within the range from 0 to 90 degrees C. The compound seemed to associate cooperatively. The thermodynamic parameters of polymerization change with temperature, so that its free enthalypy deltaG = 0.0797 T2 + 45.4 T - 7893. The dimerization enthalpy and entropy are approximately temperature-independent (deltaH2 = -4.17 kcal/mol deltas2 = -10.9 e.u.), although the function: delta g2 = -0.0308T2 +30.3T - 7213 fits experimental points better. The observed dependences can be explained by the increasing role of the hydrophobic effect with temperature and size of the aggregates. The association rate constants were determined, and a two-step reaction mechanism was demonstrated. The first step is diffusion-controlled. The second is characterized by an activation energy of approximately 2 kcal/mol and an encounter distance of approximately 8.3 angstroms.


Rewarding and aversive doses of caffeine alter activity but not conditioned place preference induced by ethanol in DBA/2J mice.
Increases in the consumption of ethanol and caffeine have been attributed to increased subjective feelings of intoxication and pleasure from the combination. Previous studies have shown that caffeine can be rewarding at low doses and aversive at high doses, although these findings are at times inconsistent between studies using comparable doses. Similarly, studies investigating the rewarding effects of ethanol and caffeine combinations have yielded mixed results. To address this issue, the present experiments were designed to investigate the rewarding effects of caffeine, as well as of caffeine + ethanol combinations. Male DBA/2J mice were exposed to an unbiased conditioned place preference (CPP) procedure with various doses of caffeine (1, 3, 10, 30 mg/kg) and ethanol (1, 2 g/kg), as well as various conditioning trial durations (5, 30, 60 min). Caffeine dose-dependently increased locomotor activity during conditioning, and produced a biphasic effect on place conditioning. Specifically, a low dose of caffeine (3 mg/kg) produced place preference, while a high dose (30 mg/kg) produced place aversion. When combined with alcohol, caffeine dose-dependently increased ethanol's stimulatory effect. However, the addition of caffeine had no effect on ethanol place preference, as there were no differences in the strength of place preference between mice conditioned with ethanol alone, and mice conditioned with any combination of ethanol and caffeine. These studies add evidence for caffeine's biphasic effects while also emphasizing the importance of considering temporal and methodological parameters when using Pavlovian conditioning procedures to study drug combinations.


Cineradiographic assessment of Björk-Shiley aortic and mitral prosthetic heart valves.
A non-invasive method for assessment of Björk-Shiley mitral and aortic heart valve movements has been developed. The Björk-Shiley tilting disc valve is described and its placement within the heart is explained. Twenty patients have been studied post-operatively with 23 valves, 10 mitral valves and 13 aortic valves, three patients having both. The method of assessment is described and consists of projecting the valve with its opaque disc marker in true profile and en face using the C arm screening system of a C.G.R. Angiorama coronary angiography unit. A cineradiograph is taken and the maximum opening angle of the valve is measured from the projected image directly or calculated from measurements taken from the image. Movement of the valve ring can also be assessed when paraprosthetic leak is suspected. The results of the study show that: (1) This method is simple, non-invasive, requires no preparation and is reproducible. (2) The valve discs open to a smaller angle than expected in most cases, and in only a few cases to 60 degrees or more. (3) It is valuable to record valve function post-operatively so that a comparison can be made later should valve malfunction be suspected. (4) In only one case was it found impossible to assess valve movement because of orientation of the valve within the patient.


Α 78-year-old female who presents with a non-resolving pneumonia: what is your diagnosis?
Non-resolving pneumonia with a mass-like radiological appearance raises clinical suspicion of many causes, with malignancy the most prominent. Infectious causes should be investigated thoroughly as they carry a better prognosis. http://ow.ly/QEPL30mLIk4.


Acute pancreatitis in children with acute lymphoblastic leukemia correlates with L-asparaginase dose intensity.
L-Asparaginase (L-Asp) is an important therapeutic for childhood acute lymphoblastic leukemia (ALL). Asparaginase-associated pancreatitis (AAP) is a severe complication of L-Asp related to the dosage. We investigated the incidence of, and risk factors for, AAP in pediatric patients with ALL. From January 2002 to December 2018, pediatric patients with ALL treated at National Taiwan University Hospital were enrolled in this study. The diagnosis of AAP was based on the criteria of the Ponte di Legno Toxicity Working Group. Of the 353 patients enrolled in this study, 14 (4.0%) developed AAP. The incidence of AAP in ALL patients was significantly higher after treatment with the 2013 protocol compared with the 2002 protocol of the Taiwan Pediatric Oncology Group (9.5% vs. 1.3%). Multivariate analysis showed that a high peak L-Asp dose intensity (>45,000 U/m2/month) and older age at diagnosis (>6.8 years) were independently predictive of AAP development. The incidence of acute pancreatitis in childhood ALL was correlated more strongly with the peak dose intensity than with the cumulative dose of L-Asp. These results could be used to reduce the treatment-related complications of ALL. L-Asparaginase is an important therapeutic for childhood acute lymphoblastic leukemia, and the accumulated dosage of L-asparaginase is considered as a major risk factor of asparaginase-associated pancreatitis. This article demonstrated that the incidence of pancreatitis correlates with the dose-intensity of L-asparaginase, but not the accumulated dosage. Identification of patient group with high risk of pancreatitis could lead to early diagnosis and reduce the complication. This finding could aid in developing further new protocol or therapeutic strategy design to reduce treatment-related complications and improve clinical outcomes of childhood acute lymphoblastic leukemia.


Dynamic expression of adenylate kinase 2 in the hippocampus of pilocarpine model rats.
Studies have shown that adenylate kinase 2 (AK2) is released from the mitochondrial inner membrane space during neuronal apoptosis, which plays an important role in temporal lobe epilepsy (TLE). We investigated the expression of AK2 in the hippocampus of a pilocarpine model rats. Using reverse transcription polymerase chain reaction and immunohistochemistry, AK2 mRNA and immune-positive cells were investigated during the entire epileptic process in pilocarpine-induced rat model of TLE. AK2 mRNA level was increased in rat hippocampus during different phases of the epileptogenesis, and reached a peak at 72 h. At 72 h time point, AK2 mRNA level was more than threefold comparing with the control. AK2-positive cells were observed in all regions of the hippocampus in model rats, but not in brain tissues of controls. The mean percentage of AK2-positive cells was increased as early as 6 h following seizure and reached a peak at 72 h. The pattern of AK2 expression over time was similar to that observed during neuronal apoptosis as detected with TUNEL staining. These results suggest that AK2 participates in the pathophysiological process of TLE and may be a marker for neuronal apoptosis induced by pathological injury in TLE.


Dynamic secondary degeneration in the spinal cord and ventral root after a focal cerebral infarction among hypertensive rats.
Cerebral infarction can cause secondary damage to nonischemic brain regions. However, whether this phenomenon will appear in central nervous system regions outside the brain remains unclear. Here we investigated pathological changes in the spinal cord and ventral root after ischemic stroke. All rats exhibited apparent neurological deficits post-MCAO, which improved gradually but could still be detected 12-weeks. Neuronal filaments in the corticospinal tract (CST) and neurons in the ventral horn were significantly declined in the contralateral cervical and lumbar enlargement 1-week post-MCAO. These decreases remained stable until 12-weeks, accompanied by progressively increased glial activation in the ventral horn. Axonal degeneration and structural derangement were evident in the contralateral cervical and lumbar ventral root 1-week post-MCAO; these changes spontaneously attenuated over time, but abnormalities could still be observed 12-weeks. The number of neural fibers in the contralateral CST and neurons in the contralateral ventral horn were positively correlated with neurological scores 12-weeks post-MCAO. Additionally, GFAP(+)cell density in the contralateral CST and ventral horn was negatively correlated with neurological scores. Our results suggest that cerebral infarction can elicit secondary degeneration in the cervical and lumbar spinal cord, as well as the projecting ventral root, which may hamper functional recovery after stroke.


Spontaneous mutagenesis: the roles of DNA repair, replication, and recombination.
There appears to be no dearth of mechanisms to explain spontaneous mutagenesis. In the case of base substitutions, data for bacteriophage T4 and especially for E. coli and S. cerevisiae suggest important roles in spontaneous mutagenesis for the error-prone repair of DNA damage (to produce mutations) and for error-free repair of DNA damage (to avoid mutagenesis). Data from the very limited number of studies on the subject suggest that about 50% of the spontaneous base substitutions in E. coli, and perhaps 90% in S. cerevisiae are due to error-prone DNA repair. On the other hand, spontaneous frameshifts and deletions seem to result from mechanisms involving recombination and replication. Spontaneous insertions have been shown to be important in the strongly polar inactivation of certain loci, but it is less important at other loci. Perhaps with continued study, the term "spontaneous mutagenesis" will be replaced by more specific terms such as 5-methylcytosine deamination mutagenesis, fatty acid oxidation mutagenesis, phenylalanine mutagenesis, and imprecise-recombination mutagenesis. While most studies have concentrated on mutator mutations, the most conclusive data for the actual source of spontaneous mutations have come from the study of antimutator mutations. Further study in this area, perhaps along with an understanding of chemical antimutagens, should be invaluable in clarifying the bases of spontaneous mutagenesis.


Coil spring fiducial markers placed safely using navigation bronchoscopy in inoperable patients allows accurate delivery of CyberKnife stereotactic radiosurgery.
CyberKnife stereotactic body radiosurgery is a potentially curative option for medically inoperable Stage I lung cancer. Fiducial marker placement in or near the tumor is required. Transthoracic placement using computed tomography guidance has been associated with a high risk of iatrogenic pneumothorax. Electromagnetic navigation bronchoscopy offers a safer method of placing markers; however, previous studies using linear markers have shown at least a 10% dislocation rate. We describe the use of coil-spring fiducial markers placed under moderate sedation in an outpatient bronchoscopy suite. A total of 52 consecutive nonoperative patients with isolated lung tumors underwent fiducial placement using electromagnetic navigation bronchoscopy. Of the 52 patients, 4 received 17 linear fiducial markers, and 49 patients with 56 tumors received 217 coil-spring fiducial markers. The procedures were considered successful if the fiducial markers had been placed in or near the tumors and had remained in place without migration, allowing radiosurgery without the need for additional fiducial markers. A total of 234 fiducial markers were successfully deployed in 52 patients with 60 tumors (mean diameter 23.7 mm). Of these 60 tumors, 35 (58%) were adjacent to the pleura. At CyberKnife planning, 8 (47%) of 17 linear fiducial markers and 215 (99%) of 217 coil-spring fiducial markers (P = .0001) were still in place. Of the 4 patients with linear fiducial markers, 2 required additional fiducial placements; none of the patients with coil fiducial markers required additional procedures. Three pneumothoraces (5.8%) occurred in peripheral lesions (2 were treated with a pig-tail chest tube and 1 with observation only). Deployment of coil spring fiducial markers using navigation bronchoscopy can safely be performed with the patient under moderate sedation with almost no migration and a 5.8% rate of pneumothorax.


[Interleukin 6 in neutrophil cultures and mononuclear cells in blood serum of patients with Lyme disease].
The aim of this study was to estimate the release of IL-6 by human neutrophils (PMN) and peripheral blood mononuclear cells (PBMC) in patients with Lyme disease confronted with the serum levels. The cells were isolated from whole blood of 25 patients and of 10 healthy donors and cultured in the presence of LPS. In the culture supernatants and serum the concentration of IL-6 with ELISA (BioSource) were measured. In patients we observed higher values of IL-6 released by unstimulated PMN and PBMC in compared with control. In contrast to control, we didn't observe increased the release of IL-6 by LPS-stimulated PMN and PBMC as compared to unstimulated cells. In the serum of patient we found increased the concentration of IL-6. The higher ability of PMN and PBMC from patients with Lyme disease to release of IL-6 and the lack response to additional stimulation indicate the activation of PMN and PBMC in vivo.


[Genetic study of bacteriophage phi81. II. Gene localization in the right arm of the phage chromosome and a comparison of phage phi81 with phages lambda and phi80 in regard to gene functions].
Data on genetic investigation on lambdoid bacteriophage phi81 made possible to localize the cos site on the prophage genetic map. Four essential genes and the gene c1 are located in the right arm of the phage genetic map. Regulatory genes of phage phi81 are found to be uncapable of functional substitution of lambda phages regulatory genes N and Q. It is discovered that some late genes of phage phi80 can be substituted with respective phage phi81 genes. No substitution possibility was observed for a number of early genes of phage phi80.


Updated cost-effectiveness and risk-benefit analysis of two infant rotavirus vaccination strategies in a high-income, low-endemic setting.
Since 2013, a biennial rotavirus pattern has emerged in the Netherlands with alternating high and low endemic years and a nearly 50% reduction in rotavirus hospitalization rates overall, while infant rotavirus vaccination has remained below 1% throughout. As the rotavirus vaccination cost-effectiveness and risk-benefit ratio in high-income settings is highly influenced by the total rotavirus disease burden, we re-evaluated two infant vaccination strategies, taking into account this recent change in rotavirus epidemiology. We used updated rotavirus disease burden estimates derived from (active) surveillance to evaluate (1) a targeted strategy with selective vaccination of infants with medical risk conditions (prematurity, low birth weight, or congenital conditions) and (2) universal vaccination including all infants. In addition, we added herd protection as well as vaccine-induced intussusception risk to our previous cost-effectiveness model. An age- and risk-group structured, discrete-time event, stochastic multi-cohort model of the Dutch pediatric population was used to estimate the costs and effects of each vaccination strategy. The targeted vaccination was cost-saving under all scenarios tested from both the healthcare payer and societal perspective at rotavirus vaccine market prices (€135/child). The cost-effectiveness ratio for universal vaccination was €51,277 at the assumed vaccine price of €75/child, using a societal perspective and 3% discount rates. Universal vaccination became cost-neutral at €32/child. At an assumed vaccine-induced intussusception rate of 1/50,000, an estimated 1707 hospitalizations and 21 fatal rotavirus cases were averted by targeted vaccination per vaccine-induced intussusception case. Applying universal vaccination, an additional 571 hospitalizations and < 1 additional rotavirus death were averted in healthy children per vaccine-induced intussusception case. While universal infant rotavirus vaccination results in the highest reductions in the population burden of rotavirus, targeted vaccination should be considered as a cost-saving alternative with a favorable risk-benefit ratio for high-income settings where universal implementation is unfeasible because of budget restrictions, low rotavirus endemicity, and/or public acceptance.


Squamous cell carcinoma of the nasal columella: a retrospective study of 66 cases from the GETTEC.
Squamous cell carcinoma of the nasal columella (SCCNC) is a rare disease. We aimed to define a strategy for the diagnosis and management of nasal columella squamous cell carcinoma. Medical records of 66 patients presenting with columella squamous cell carcinoma in nine French hospitals, from 1980 to 2003, were evaluated to determine the clinical characteristics and current treatment of the disease. Mean age was 69 years. The sex ratio was one female for three males. Majority of the lesions were T1 N0 according to the classification international union against cancer. Patients underwent one of the three treatments: surgery alone for the T1 lesions, radiotherapy for tumors of T2-T3 and combined (surgery and radiotherapy) for T4 lesions. The 5-year Kaplan-Meier survival test was 39% and no difference was found between the therapeutic groups. Thirty-five (53%) tumor recurrences were observed with a median time of 43 months. As regards TNM classification and treatment, no significant difference between the two groups disease free/recurrence was found. Prognosis of early lesions was considered better than the advanced lesions. SCCNC is difficult to manage and has a poor prognosis. No therapeutic solution has yet been confirmed in the treatment of this pathology.


Technique for fabrication of the mandibular denture over the staple bone implant using a permanent heat-cured base.
A technique that uses processed bases to which the semiprecision attachments are attached intraorally has been described. The teeth are cured in heat-cured resin directly to the base. Advantages of using heat-cured processed bases are that (1) the base is accurately related to the attachments; (2) there is direct access to the attachments for joining them to the base; (3) extension, retention, and stability can be evaluated prior to completion of the dentures; and (4) accurate jaw relation recordings can be made due to the accuracy of the fit of the bases.


Ensuring fair, safe, and interpretable artificial intelligence-based prediction tools in a real-world oncological setting.
Cancer patients often experience treatment-related symptoms which, if uncontrolled, may require emergency department admission. We developed models identifying breast or genitourinary cancer patients at the risk of attending emergency department (ED) within 30-days and demonstrated the development, validation, and proactive approach to in-production monitoring of an artificial intelligence-based predictive model during a 3-month simulated deployment at a cancer hospital in the United States. We used routinely-collected electronic health record data to develop our predictive models. We evaluated models including a variational autoencoder k-nearest neighbors algorithm (VAE-kNN) and model behaviors with a sample containing 84,138 observations from 28,369 patients. We assessed the model during a 77-day production period exposure to live data using a proactively monitoring process with predefined metrics. Performance of the VAE-kNN algorithm is exceptional (Area under the receiver-operating characteristics, AUC = 0.80) and remains stable across demographic and disease groups over the production period (AUC 0.74-0.82). We can detect issues in data feeds using our monitoring process to create immediate insights into future model performance. Our algorithm demonstrates exceptional performance at predicting risk of 30-day ED visits. We confirm that model outputs are equitable and stable over time using a proactive monitoring approach.


Dermatoscopy of Common Lesions in Pediatric Dermatology.
The use of dermatoscopy to assist in the diagnosis of a variety of proliferative, pigmentary, inflammatory, infectious, congenital, and genetic cutaneous and skin appendage disorders is constantly increasing, as it is effective, affordable, noninvasive, and quick to perform.


West nile poliomyelitis in a 7-year-old child.
West Nile poliomyelitis is a well-described neurologic manifestation of West Nile viral infection in adults. However, few reports have described this manifestation in children infected with West Nile virus. We describe a 7-year-old boy who developed West Nile poliomyelitis with flaccid paralysis of his left leg. Electrodiagnostic testing and radiologic imaging confirmed anterior horn cell injury. We report on his course clinically and electrodiagnostically over 20 months.


Improving Transfer of Learning: An Innovative Comentoring Program to Enhance Workplace Implementation After an Occupational Therapy Course on Autism Spectrum Disorders.
Research suggests that learning gained through training is infrequently implemented in the workplace. A short-term postcourse comentoring program was developed with the aim of facilitating workplace implementation of learning after a 3-day course for occupational therapists. The program was evaluated for usefulness, successes, challenges, recommended improvements, and associations with changes in self-rated knowledge and confidence. Two months after the course, 42 participants completed an evaluation of the comentoring program with closed- and open-ended questions addressing usefulness, successes, challenges, pairing preferences, and recommendations. They also completed a record on whether or not they had worked on goals nominated in their comentoring contract. Before and 2 months after the course, they completed a self-rated questionnaire on knowledge and confidence. The comentoring program was recommended by 80% of participants. Benefits included opportunities for information and resource sharing, debriefing, problem solving, reassurance, and implementation of ideas. Ninety-five percent of participants worked on some or all their comentoring goals. Although there were significant improvements in knowledge (P < 0.001) and confidence (P < 0.001), the total comentoring evaluation score was not significantly associated with these changes. It is therefore possible that these improvements related to the course itself rather than the comentoring program. Challenges related to time, scheduling, distance, and pairing of comentors. Reported benefits of the program included enhanced psychosocial support and prompting to trial newly learned strategies. Effectiveness may be improved by setting aside time for comentoring in the workplace and better matching of comentors.


Sweet-like syndrome and multiple COVID arm syndrome following COVID-19 vaccines: 'specific' patterns in a series of 192 patients.
The two clinico-pathological patterns are 'Sweet-like syndrome' and 'Multiple COVID-Arm'. 'Sweet-like syndrome' presents clinically as erythematous and oedematous papules or plaques, sometimes developing vesiculation or bullae. Histology shows classical Sweet syndrome with a diffuse dermal neutrophilic infiltrate, or an infiltrate of histiocyte-like immature myeloid cells consistent with a histiocytoid Sweet syndrome. 'Multiple COVID-arm' is characterized by multiple large inflammatory plaques with histological analyses showing a perivascular and interstitial inflammatory infiltrate with eosinophils.


Supporting Veteran's Administration Medical Center Directors' Decisions When Adopting Innovative Practices: Development and Implementation of the "QuickView" and "WishList" Tools.
Background Since 2015, the Veterans Health Administration (VHA) Diffusion of Excellence Program has supported spread of practices developed by frontline employees. Shark Tank-style competitions encourage "Sharks" nationwide (VHA medical center/regional directors) to bid for the opportunity to implement practices at their institutions. Methods The authors evaluated bidding strategies (2016-2020), developing the "QuickView" practice comparator to promote informed bidding. Program leaders distributed QuickView and revised versions in subsequent competitions. Our team utilized in-person observation, online chats after the competition, bidder interviews, and bid analysis to evaluate QuickView use. Bids were ranked based on demonstrated understanding of resources required for practice implementation. Results Sharks stated that QuickView supported preparation before the competition and suggested improvements. Our revised tool reported necessary staff time and incorporated a "WishList" from practice finalists detailing minimum requirements for successful implementation. Bids from later years reflected increased review of facilities' current states before the competition and increased understanding of the resources needed for implementation. Percentage of bids describing local need for the practice rose from 2016 to 2020: 4.7% (6/127); 62.1% (54/87); 78.3% (36/46); 80.6% (29/36); 89.7% (26/29). Percentage of bids committing specific resources rose following QuickView introduction: 81.1% (103/127) in 2016, 69.0% (60/87) in 2017, then 73.9% (34/46) in 2018, 88.9% (32/36) in 2019, and 89.7% (26/29) in 2020. Discussion In the years following QuickView/WishList implementation, bids reflected increased assessment before the competition of both local needs and available resources. Conclusion Selection of a new practice for implementation requires an understanding of local need, necessary resources, and fit. QuickView and WishList appear to support these determinations.


Presence of a non-sulphated glucosaminoglycan in embryonic cornea.
An analtical evidence has been obtained for the presence of a non-sulphated glucosaminoglycan in the young embryonic White Leghorn chicken cornea, consisting of nearly equimolar amounts of glucosamine and glucuronic acid. The polysaccharide is supposed to be identical with hyaluronic acid.


Has price competition changed hospital revenues and expenses in New York?
This study analyzes the factors that influenced hospital expenses and revenues prior to and following the enactment of the New York State Health Care Reform Act of 1996 (HCRA)-the period from 1994-1999. HCRA was expected to encourage price competition which in turn was anticipated to lower hospital revenues and expenses. We measured the differential effects on hospital revenues and expenses in markets with varying degrees of competition. We also measured the relationship between hospital revenues and expenses and the increased concentration resulting from the formation of local hospital systems. We found that revenues and expenses both grew more slowly for hospitals located in more competitive markets; hospital systems that increased concentration tended to have higher revenues. In the short run at least, price competition induced by HCRA did constrain both hospital expense and revenue growth, although the increase in hospital mergers countered this trend.


The effect of time delay on infection in open long-bone fractures: a 5-year prospective audit from a district general hospital.
To determine whether a time delay of greater than 6 h from injury to surgical debridement affected the infection rate in open long-bone fractures in a typical district general hospital in the UK. Prospective audit over 5 years. 142 open long-bone fractures in 130 patients were seen over a 5-year period between 1996 and 2001 in our hospital. 115 fractures in 103 patients were available for study. The data were collected prospectively in weekly audit meetings. Patients were followed until clinical or radiological union occurred or until a secondary procedure for non-union or infection was carried out. Surgical debridement was carried out in less than 6 h from injury in 60% of cases and in greater than 6 h from injury in 40% of cases. Infection rates were 10.1% and 10.8%, respectively. We could not demonstrate a significantly increased infection rate in patients in whom surgical debridement occurred greater than 6 h after injury.


EGFR tyrosine kinase activity and Rab GTPases coordinate EGFR trafficking to regulate macrophage activation in sepsis.
EGFR phosphorylation is required for TLR4-mediated macrophage activation during sepsis. However, whether and how intracellular EGFR is transported during endotoxemia have largely been unknown. Here, we show that LPS promotes high levels cell surface expression of EGFR in macrophages through two different transport mechanisms. On one hand, Rab10 is required for EEA1-mediated the membrane translocation of EGFR from the Golgi. On the other hand, EGFR phosphorylation prevents its endocytosis in a kinase activity-dependent manner. Erlotinib, an EGFR tyrosine kinase inhibitor, significantly reduced membrane EGFR expression in LPS-activated macrophage. Mechanistically, upon LPS induced TLR4/EGFR phosphorylation, MAPK14 phosphorylated Rab7a at S72 impaired membrane receptor late endocytosis, which maintains EGFR membrane localization though blocking its lysosomal degradation. Meanwhile, Rab5a is also involved in the early endocytosis of EGFR. Subsequently, inhibition of EGFR phosphorylation switches M1 phenotype to M2 phenotype and alleviates sepsis-induced acute lung injury. Mechanistic study demonstrated that Erlotinib suppressed glycolysis-dependent M1 polarization via PKM2/HIF-1ɑ pathway and promoted M2 polarization through up-regulating PPARγ induced glutamine metabolism. Collectively, our data elucidated a more in-depth mechanism of macrophages activation, and provided stronger evidence supporting EGFR as a potential therapeutic target for the treatment of sepsis.


Detecting changes in patient treatment satisfaction with medicines: the SATMED-Q.
The Treatment Satisfaction with Medicines Questionnaire (SATMED-Q) has shown appropriate psychometric properties exploring patients' satisfaction with treatment. Responsiveness (sensitivity to change) and known-group validity, however, still remained unknown. Thus, the goal of this study was to explore such psychometric properties for the SATMED-Q. We used data from a 6-month prospective study carried out in pain clinics, which included patients with chronic refractory pain of neuropathic origin who needed a change in their therapies. Sensitivity to change was assessed by comparing changes in the total and domain scores between baseline and end-of-trial visits according to patients' response criterion: pain reduction 50% or more (responder). Also, correlations between changes in pain intensity and satisfaction scores were computed. Known-groups validity was explored by comparing the degree of satisfaction between groups of different levels of pain. The sample was formed with 728 subjects (57.8 years, 61.0% women). After changing their therapy, 47% of the patients were considered responders, and pain intensity was reduced by an average of 42.9%±32.4% (P<0.001), which was significantly correlated (r =-0.524, P<0.001) with total treatment satisfaction improvement. The total score in the SATMED-Q was significantly higher in responders than in nonresponders: 80.9 (79.6-82.3) versus 66.5 (65.0-98.0) (P<0.001). Also, the instrument showed different scores of satisfaction according to different degrees of pain (mild, moderate, severe), F = 116.8 (P<0.001) in the total composite score and domains. The SATMED-Q was sensitive to changes in patients' satisfaction with treatment. In addition, patients' different heath statuses are correlated with different levels of satisfaction with treatment.


Trends of inflammatory bowel disease at a tertiary care center in northern India.
Inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), is increasingly being reported from India and other Asian countries. This study looks into the changing trends of IBD at a tertiary care center in north India over last two decades. Retrospective analysis of a prospectively maintained database of patients diagnosed with IBD between January 1991 and December 2015 was conducted. The study period was divided into five times cohorts (1991-1995, 1996-2000, 2001-2005, 2006-2010, 2011-2015). During the study period, 2,467 patients (UC [n = 2,137, 86.6%], CD [n = 330, 13.3%], mean age 38.5 ± 13.3 years; 55.9% males) were registered. The proportion of patients with CD increased (ratio of UC to CD declined from 15.7:1 to 4:1). The mean age at diagnosis decreased for UC (45.7 ± 12.1 years in 1991-1995 vs. 37.6 ± 13.0 years in 2011-2015; P= 0.001) and remained consistent for CD (41.3 ± 13.6 years in 1996-2000 vs. 41.3 ± 16.9 years in 2011-2015, P= 0.86). Patients with proctitis in UC and isolated ileal disease in CD increased over the study period (P= 0.001 and P= 0.007, respectively). Inflammatory CD increased (P= 0.009) whereas stricturing CD decreased (P= 0.01) across all cohorts. There was a trend towards less severe presentation of both UC and CD. The use of thiopurines (P= 0.02) and biologics increased (P= 0.001) with no significant change in trends for requirements of surgery (P= 0.9). Increasing prevalence of CD, younger age at diagnosis, diagnosis at an earlier and milder stage, greater use of thiopurines and biologics were observed.


Growth/no growth interfaces of table olive related yeasts for natamycin, citric acid and sodium chloride.
The present work uses a logistic/probabilistic model to obtain the growth/no growth interfaces of Saccharomyces cerevisiae, Wickerhamomyces anomalus and Candida boidinii (three yeast species commonly isolated from table olives) as a function of the diverse combinations of natamycin (0-30 mg/L), citric acid (0.00-0.45%) and sodium chloride (3-6%). Mathematical models obtained individually for each yeast species showed that progressive concentrations of citric acid decreased the effect of natamycin, which was only observed below 0.15% citric acid. Sodium chloride concentrations around 5% slightly increased S. cerevisiae and C. boidinii resistance to natamycin, although concentrations above 6% of NaCl always favoured inhibition by this antimycotic. An overall growth/no growth interface, built considering data from the three yeast species, revealed that inhibition in the absence of citric acid and at 4.5% NaCl can be reached using natamycin concentrations between 12 and 30 mg/L for growth probabilities between 0.10 and 0.01, respectively. Results obtained in this survey show that is not advisable to use jointly natamycin and citric acid in table olive packaging because of the observed antagonistic effects between both preservatives, but table olives processed without citric acid could allow the application of the antifungal.


Mini-percutaneous nephrolithotomy with high-power holmium YAG laser in pediatric patients with staghorn and complex calculi.
Shock wave lithotripsy (SWL) is the treatment of choice for most cases of renal lithiasis in children. Some cases, however, are refractory to SWL and may be associated with anatomic and metabolic changes or a large stone burden. In these circumstances, mini-percutaneous nephrolithotomy (mini-PCNL) is an option. The aim was to assess the safety and efficacy of high-power holmium YAG (Ho:YAG) laser in mini-PCNL for staghorn calculi. The clinical records relating to 35 mini-PCNLs (Table) performed between January 2008 and December 2012 in 33 patients (27 boys and 6 girls; mean age 7 years, range 2-18 years) were retrospectively reviewed. Twenty-two (66.7%) of the patients had undergone a previous SWL (28.6% three sessions). The mini-PCNL puncture technique used involved fluoroscopic guidance with the C arm at 0-90° in the supine position. An 18F tract was used. Stone fragmentation was performed with a high-power Ho:YAG laser (Lumenis 100 W). Ten of the mini-PCNLs (28.6%) were right sided, and 25 were left sided (71.4%). Stones were located in the lower calyceal group in 64% of patients and in the renal pelvis in 50%. The mean stone size was 4.46 cm(2) (range 3-13.20 cm(2)). The number of stones varied between one and 20, and 83.3% were radiopaque. The laser was set at 70 W (range 50-100 W) (3.5 J/pulse with a frequency of 20 pulses/s). The mean surgical time was 150 min. In 78% of patients, complete stone clearance was achieved, and the overall stone-free rate rose to 85% when residual stones were treated with either SWL or retrograde intrarenal surgery. No perioperative complications were seen. There are few reports in the literature concerning the use of a high-power laser for treatment of complex stones and the few that do exist relate to adults have similar results, showing it to be highly effective and safe, with low morbidity. Some limitations of the present study must be acknowledged. It was retrospective and a relatively small number of patients were included. However, randomized prospective studies are required to confirm our findings and conclusions. In the pediatric population, mini-PCNL with high-power Ho:YAG laser is feasible and safe and represents the technique of choice for complex renal stones.


Carbon dioxide digital subtraction arteriography: a pilot study.
To investigate the diagnostic value of carbon dioxide arteriograms in patients with peripheral vascular disease, ten patients in whom standard contrast arteriography was contraindicated underwent carbon dioxide digital subtraction arteriography. Lower extremity ischemia or severe hypertension with renal insufficiency were the indications for arteriography. Standard contrast arteriography was precluded by chronic nondialysis-dependent renal insufficiency, severe congestive heart failure or contrast hypersensitivity. All critical arterial segments were well visualized with the exception of the infrapopliteal arterial tree in three patients. Adequate imaging of this segment required the addition of 20 cc of dilute nonionic contrast. Guided by carbon dioxide digital subtraction arteriography, four percutaneous transluminal angioplasties and three reconstructive procedures were successfully performed. One patient did not have surgically reconstructible disease and two had renal arteries without critical stenoses. Renal function transiently deteriorated in one patient who received 20 cc of nonionic contrast. No adverse events occurred due to carbon dioxide. Clinically useful diagnostic arteriograms are possible using carbon dioxide as the contrast agent.


Neonatal plantar response revisited.
To evaluate plantar response in the early neonatal period in normal, term, healthy newborn infants. This was a prospective study set in the postnatal ward of a tertiary care hospital. The plantar response was elicited in 256 healthy, term, appropriate-for-gestational-age neonates during their first 7 days of life, utilizing the thumb-nail-drag method. The response was tested daily at intervals of 24 h, until the neonate was discharged. A total of 597 observations were made, and the responses were classified as extensor, flexor or equivocal. The overall plantar response was found to be predominantly extensor (73.8%), followed by equivocal (17.3%) and, finally, flexor (8.9%). The plantar response was bilaterally extensor in 72.5%, bilaterally equivocal in 14.2%, and bilaterally flexor in 7.7% of observations, respectively. Asymmetrical plantar response was elicited in 5.4% of observations. No difference was observed in individual categories based on age (<12 h, 12.1-24 h, 24.1-72 h, 72.1 h-7 days) and site of response (right/left foot). The plantar response in healthy, term neonates is predominantly extensor. Further, the relatively high frequency of asymmetrical and symmetrical flexor responses limits its clinical usefulness in the early neonatal period.


Familial inversion translocation (8;13) with partial trisomy 13 in several family members.
A partial trisomy 13q was observed in siblings with hexadactylia, hypertelorism, hemangioma and severe psychomotor retardation. It originated from a maternal inversion translocation 46,XX,inv(8)(q23q241),t(8;13)(q241;q32). The family showed a pedigree pattern typical for the segregation of a chromosomal translocation. In spite of this the diagnosis was delayed several years, because the bands involved from the two chromosomes were of great similarity. This stresses the importance of reinvestigating families with a clinical suspicion of a chromosomal syndrome, preferentially with prometaphase chromosomes. The identification of a chromosomal rearrangement is essential for genetic counselling and prenatal diagnosis.


Fatal hemorrhage after radiochemotherapy for leptomeningeal dissemination of glioma: report of two cases.
Dissemination of malignant gliomas is often observed, but the treatment of choice for leptomeningeal dissemination has not been established. A 55-year-old man suffered fatal hemorrhage 1 day after whole brain irradiation (2 Gy) along with intravenous administration of 150 mg of nitrosourea (ACNU) for leptomeningeal dissemination of glioblastoma. A 14-year-old boy suffered hemorrhage after intrathecal administration of 10 mg of methotrexate for recurrence of leptomeningeal dissemination of anaplastic astrocytoma 4 months after local brain irradiation and ACNU administration, and died 19 days later. Massive fatal hemorrhage is an unusual but possible acute complication after radiochemotherapy for leptomeningeal dissemination of gliomas.


Bone Age Assessments by Quantitative Ultrasound (SonicBone) and Hand X-ray Based Methods are Comparable.
Bone maturation is currently assessed by subjective and automated radiography. To evaluate the concordance and reproducibility of a quantitative ultrasound (QUS) based device versus X-ray based methods. The study population comprised 150 children, 76 males, 4-17 years of age. X-ray scans were evaluated according to wrist, carpal and phalanx areas for bone age. QUS was performed by the the BAUS™ device (SonicBone, Rishon Lezion, Israel), using speed-of-sound (SOS) and distance attenuation factor (ATN) in similar areas. Data from 100 subjects were used to establish the device conversion equation, and 50 measurements were assigned to assess inter-modality agreement. BAUS showed high repeatability performance, 0.73% relative standard deviation for SOS and 3.5% for ATN. R2 for the conversion equation, including gender, SOS, and ATN, was 0.80 for all methods (P < 0.001). There was no significant bias in bone age assessments. Bone age assessment by SonicBone is comparable to the assessment by X-ray based methods.


Nano-level monitoring of Er(III) by fabrication of coated graphite electrode based on newly synthesized Schiff base as neutral carrier.
Plasticized membranes using N-(-3-((thiazol-2-ylimino)methyl)benzylidene)thiazol-2-amine (S1) and 5-((-3-((5-mercapto-1,3,4-thiadiazol-2-ylimino)methyl)benzylidene)amino)-1,3,4-thiadiazole-2-thiol (S2) have been prepared and explored as Er (III) selective electrodes. Effect of various plasticizers viz. dibutylphthalate, tri-n-butylphosphate, dioctylphthalate, acetophenone, 1-chloronapthalene, o-nitrophenyloctylether, and anion excluders viz. sodium tetraphenylborate and potassium tetrakis-p-(chlorophenyl)borate was studied in detail and improved performance was observed. Optimum performance was observed for the membrane electrode having a composition of S2: PVC: o-NPOE: KTpClPB in the ratio of 4: 38: 55: 3 (w/w, mg). The performance of the PME based on S2 was compared with CGE. The electrodes exhibit Nernstian slope for Er (III) ion with detection limit 5.4 × 10(-8)mol L(-1) for PME and 6.1 × 10(-9)mol L(-1) for CGE. The response time for PME and CGE was found to be 12s and 9s respectively. The practical utility of the CGE has been demonstrated by its usage as an indicator electrode in potentiometric titration of EDTA with Er (III) solution and determination of fluoride ions in mouthwash solution. The proposed electrode was also applied to the determination of added Er(3+) ion in water and binary mixtures. It is found that the electrode could be able to recover the Er(3+) ion in 96.2-99.5%.


Histamine may contribute to vortioxetine's procognitive effects; possibly through an orexigenic mechanism.
Vortioxetine is a novel multimodal antidepressant that acts as a serotonin (5-HT)3, 5-HT7, and 5-HT1D receptor antagonist; 5-HT1B receptor partial agonist; 5-HT1A receptor agonist; and 5-HT transporter inhibitor in vitro. In preclinical and clinical studies vortioxetine demonstrates positive effects on cognitive dysfunction. Vortioxetine's effect on cognitive function likely involves the modulation of several neurotransmitter systems. Acute and chronic administration of vortioxetine resulted in changes in histamine concentrations in microdialysates collected from the rat prefrontal cortex and ventral hippocampus. Based on these results and a literature review of the current understanding of the interaction between the histaminergic and serotonergic systems and the role of histamine on cognitive function, we hypothesize that vortioxetine through an activation of the orexinergic system stimulates the tuberomammilary nucleus and enhances histaminergic neurotransmission, which contributes to vortioxetine's positive effects on cognitive function.


Predicting virulence factors of immunological interest.
In this chapter, three prediction servers used for predicting virulence factors, bacterial toxins, and neurotoxins have been described. VICMpred server predicts the functional proteins of gram-negative bacteria that include virulence factors, information molecule, cellular process, and metabolism molecule. BTXpred server allows users to predict bacterial toxins, its release, and further classification of exotoxins. NTXpred server allows prediction of neurotoxins and further classifying them based on their function and source.


Detection of monosomy 7 by fluorescence in situ hybridization in acute nonlymphocytic leukemia and myelodysplastic syndrome.
Fluorescence in situ hybridization (FISH) with a chromosome 7 specific alpha satellite DNA probe was used to detect monosomy 7 in interphase and metaphase cells obtained from patients with myelodysplastic syndrome (MDS) and acute nonlymphocytic leukemia (ANLL). Chromosome analysis revealed monosomy 7, either alone or as part of a complex chromosome abnormality, in all cell samples. FISH analyses of 12 marrow samples and a blood sample using a chromosome 7 specific alpha satellite DNA probe revealed a single fluorescence spot in 80.5-97.5% of interphase cells indicating monosomy 7. In contrast, 83.5-92.0% of the same cells had two copies of chromosome 17 as two fluorescent spots were detected using a chromosome 17 specific alpha satellite DNA probe used as a positive control. The proportion of interphase cells with monosomy 7 did not correlated with the percentage of metaphase cells with monosomy 7 detected by conventional karyotyping or with the percentage of blast cells in the bone marrow.


Leptin treatment prevents type I diabetic marrow adiposity but not bone loss in mice.
Leptin is a hormone secreted by adipocytes that is implicated in the regulation of bone density. Serum leptin levels are decreased in rodent models of type 1 (T1-) diabetes and in diabetic patients. Whether leptin mediates diabetic bone changes is unclear. Therefore, we treated control and T1-diabetic mice with chronic (28 days) subcutaneous infusion of leptin or saline to elucidate the therapeutic potential of leptin for diabetic osteoporosis. Leptin prevented the increase of marrow adipocytes and the increased aP2 expression that we observed in vehicle-treated diabetic mice. However, leptin did not prevent T1-diabetic decreases in trabecular bone volume fraction or bone mineral density in tibia or vertebrae. Consistent with this finding, markers of bone formation (osteocalcin RNA and serum levels) in diabetic mice were not restored to normal levels with leptin treatment. Interestingly, markers of bone resorption (TRAP5 RNA and serum levels) were decreased in diabetic mice by leptin treatment. In summary, we have demonstrated a link between low leptin levels in T1-diabetes and marrow adiposity. However, leptin treatment alone was not successful in preventing bone loss.


Neuropsychological follow-up of patients operated for aneurysms of anterior communicating artery.
The neuropsychological outcome of 43 patients operated for ACoA aneurysms was assessed with a battery of 15 tests, tapping a wide spectrum of cognitive abilities. As a group ACoA aneurysms patients were impaired on 8 tests, including the three assessing memory. When however, patients falling below the cut-off point determined in normal controls were considered, 42% of the sample was unimpaired and only 35% fell on two or more tests. The analysis of single cases showed that memory was often affected, but short-term memory even more than long-term memory. In addition to the well known memory disorder, the patients showed a wide range of neuropsychological defects, including language and space functions. The hypotheses that could account for this broad-based neuropsychological impairment are discussed.


Reliability of short-listing in medical student selection.
One in eight Universities Central Council on Admissions (UCCA) applications for admission to St Mary's Hospital Medical School in 1986 were in due course recirculated to the four short-listers, being seen again either by the same short-lister or by another short-lister. Intrarater reliabilities were high, not only for measures of educational achievement, but also for the more subjective assessments. Interrater reliabilities were more variable, being very high for educational achievement, but rather lower for the more subjective items, and being lowest for the assessment of 'Interests', suggesting divergence between short-listers' perceptions of the terms. Nevertheless, all reliabilities were sufficiently high to justify the continued use of these criteria during selection.


Bioaerosols--sources and control measures.
In every field of activity where organic material is being handled, emissions of dust, gases, odor as well as bioaerosols are bound to arise. For this reason, waste management facilities or else agricultural enterprises are potential emission sources of bioaerosols. The dispersion of bioaerosols from waste treatment facilities and their health impacts continue to be the subject of numerous discussions. This article addresses organizational and engineering measures for the mitigation of bioaerosol emissions. The required scale of emission reduction and the choice of microbiological parameters have to be assessed with respect to location and facility type.


NR2C and NR2D subunits of NMDA receptors in frog and turtle retina.
Glutamate NMDA (N-methyl-D-aspartate) receptors are widely distributed in the central nervous system where they are involved in cognitive processes, motor control and many other functions. They are also well studied in the retina, which may be regarded as a biological model of the nervous system. However, little is known about NR2C and NR2D subunits of NMDA receptors, which have some specific features as compared to other subunits. Consequently the aim of the present study was to investigate their distribution in frog (Rana ridibunda) and turtle (Emys orbicularis) retinas which possess mixed and cone types of retina respectively. The experiments were performed using an indirect immunofluorescence method. Four antibodies directed to NR2C and NR2D subunits of NMDA receptor, as well as three antibodies directed to different splice variants of NR1 subunit, which is known to be obligatory for proper functioning of the receptor, were applied. All antibodies caused well expressed labeling in frog and turtle retinas. The NR2C and NR2D subunits were localized in glial Müller cells, while the NR1 subunit had both neuronal and glial localization. Our results show that glial NMDA receptors differ from neuronal ones in their subunit composition. The functional significance of the NMDA receptors and their NR2C and NR2D subunits, in particular for the neuron-glia interactions, is discussed.


Mechanisms, pools, and sites of spontaneous vesicle release at synapses of rod and cone photoreceptors.
Photoreceptors have depolarized resting potentials that stimulate calcium-dependent release continuously from a large vesicle pool but neurons can also release vesicles without stimulation. We characterized the Ca(2+) dependence, vesicle pools, and release sites involved in spontaneous release at photoreceptor ribbon synapses. In whole-cell recordings from light-adapted horizontal cells (HCs) of tiger salamander retina, we detected miniature excitatory post-synaptic currents (mEPSCs) when no stimulation was applied to promote exocytosis. Blocking Ca(2+) influx by lowering extracellular Ca(2+) , by application of Cd(2+) and other agents reduced the frequency of mEPSCs but did not eliminate them, indicating that mEPSCs can occur independently of Ca(2+) . We also measured release presynaptically from rods and cones by examining quantal glutamate transporter anion currents. Presynaptic quantal event frequency was reduced by Cd(2+) or by increased intracellular Ca(2+) buffering in rods, but not in cones, that were voltage clamped at -70 mV. By inhibiting the vesicle cycle with bafilomycin, we found the frequency of mEPSCs declined more rapidly than the amplitude of evoked excitatory post-synaptic currents (EPSCs) suggesting a possible separation between vesicle pools in evoked and spontaneous exocytosis. We mapped sites of Ca(2+) -independent release using total internal reflectance fluorescence (TIRF) microscopy to visualize fusion of individual vesicles loaded with dextran-conjugated pHrodo. Spontaneous release in rods occurred more frequently at non-ribbon sites than evoked release events. The function of Ca(2+) -independent spontaneous release at continuously active photoreceptor synapses remains unclear, but the low frequency of spontaneous quanta limits their impact on noise.


Inhibition of endothelial cell adhesion and proliferation by extracellular matrix from vascular smooth muscle cells: role of type V collagen.
Endothelial cells recovering from damage due to disease or surgical procedures come into close contact with extracellular matrix (ECM) secreted by intimal vascular smooth muscle cells (VSMCs). We have investigated these relationships using human umbilical artery endothelial cells (HUAECs) and human mammary artery VSMC in vitro. HUAEC adhesion and proliferation were significantly lower on ECM secreted by VSMC compared with HUAEC ECM or surface-coated fibronectin. Characterisation of the ECM of both cell types with monoclonal antibodies showed that the ECM secreted by VSMC contained significantly more elastin, chondroitin sulphate and collagen types I, III and V than that from HUAECs. HUAECs adhered poorly to collagen type V coated on plastic and not at all to elastin. When these proteins were co-coated with fibronectin, elastin did not inhibit migration or proliferation compared to the response on fibronectin but collagen type V significantly inhibited both. Treatment of VSMC ECM with enzymes which selectively depleted the matrix of collagen types I, III and IV, or chondroitin sulphate, had no effect on HUAEC responses to the ECM, suggesting that these molecules did not contribute to the inhibition of HUAECs. Treatment of VSMC ECM with a mixture of collagenases, selectively depleted the matrix of collagen type V, as well as types I, III and IV. Such depleted ECMs supported increased proliferation of HUAECs compared to buffer controls. Overall these results suggest that collagen V secreted into the ECM of VSMC may inhibit the recovery of adjacent endothelium.


Rapid detection of haloarchaeal carotenoids via liquid-liquid microextraction enabled direct TLC MALDI-MS.
For the first time, we demonstrate the use of TiO2 nanoparticles (NPs) for enhancing the carotenoid production by the extremophilic haloarchea, Haloferax mediterranei. TiO2 NPs at optimal concentration of 375 mg/L results in a 95% increase in the production of carotenoid pigment compared to the control (no TiO2 NPs). The carotenoid pigments extracted from TiO2 NPs treated H. mediterranei cells, were separated using thin layer chromatography (TLC). The separated carotenoid spots were subjected directly for MALDI MS detection. To limit the sample diffusion during matrix addition on TLC plates, a simple bordering mode was exercised. Using this method we were able to detect the pigments successfully using MALDI-MS, directly from TLC plates after separation. In addition, we also applied the Pt NPs capped with ODT via Liquid-liquid microextraction (LLME) for extracting the pigment molecules from the halobacteria in MALDI-MS. These novel NP approaches possess numerous advantages such as; rapidity, ease in synthesis, high sensitivity and low cost.


Detection of intracellular expression and secretion of interferon-gamma at the single-cell level after activation of human T cells with tetanus toxoid in vitro.
Activation of T cells results in intracellular expression and secretion of cytokines such as interferon (IFN)-gamma. Here we have used three different assays for determination of IFN-gamma in tetanus toxoid- or mitogen-activated human T cell cultures. Two of these assays [intracytoplasmic immunofluorescence and enzyme-linked immuno spot assay (ELISPOT)] determined the expression and secretion of IFN-gamma at the single-cell level while the third assay enzyme-linked immunosorbent assay (ELISA) measured IFN-gamma secreted into the culture supernatant. Comparison of all three tests revealed a good correlation between the ELISPOT assay and the ELISA, whereas expression of intracellular IFN-gamma showed a qualitative but not a quantitative correlation with the latter. Both the immunospot assay and the immunofluorescence may be used to detect approximate numbers of specific T cells even when present at low frequencies. With the use of the immunospot assay antigen-specific T cells could be detected even in the absence of detectable IFN-gamma in the culture supernatants. However, the ELISA assay should be more convenient for screening large clinical material.


2018 tallied facial soft tissue thicknesses for adults and sub-adults.
The tallied facial soft tissue thicknesses (or T-Tables) represent grand means of published facial soft tissue thickness sample means. These sample means have been drawn from across the full-breadth of the facial soft tissue thickness (FSTT) literature, including forensic science, anthropology and odontology. The report of new summary statistics for >1290 new sub-adults and >2200 new adults since the last T-Table calculation, in 2008 for sub-adults and 2013 for adults respectively, makes their update timely. The maximum sample sizes at any landmark now stand at 3023 for individuals aged 0-11 years old (g-g'); 3145 for individuals aged 12-17 years old (n-se'); and 10,333 for adults (n-se'). Following the recalculation of grand weighted means and comparison to the original 2008 data, some shifts in the T-Table statistics are evident at specific landmarks, namely: 2-2.5mm increases at gonion (go-go') and mid-mandibular border (mmb-mmb') for adults; 3.5mm decrease at gonion (go-go') for 12-17year olds; and 2.0mm decrease at menton (me-me') for 0-11year olds. Differences at all other landmarks (91-100% depending on the dataset) were minimal being <1.0mm. Performance tests of the new grand means as point estimators (using individuals with known FSTT size from the C-Table), show the 2018 T-Table statistics to produce marginally less error than the 2013 means: 2018 standard error of the estimate=3.7mm in contrast to 2013 standard error of the estimate=3.9mm. The long run nature of the T-Table statistics (i.e., big data) and quantified performance test accuracies on known subjects, earmark the 2018 T-Table as the premier FSTT standard for craniofacial identification casework. In the distant future, this is likely to change as the C-Table raw data repository grows, allowing shorths and shormaxes to be calculated for large samples. Given current raw data repository sample sizes of 0-1574 for T-Table landmarks (notably lower for younger individuals), there is some way to go before enhanced central tendency estimators can entirely replace untrimmed arithmetic means.


A numerical study of the influence of ammonia addition on the auto-ignition limits of methane/air mixtures.
In this study the auto-ignition limit of ammonia/methane/air mixtures is calculated based upon a perfectly stirred reactor model with convective heat transfer. The results of four different reaction mechanisms are compared with existing experimental data at an initial temperature of 723 K with ammonia concentrations of 0-20 mol.% and methane concentrations of 2.5-10 mol.%. It is found that the calculation of the auto-ignition limit pressure at constant temperature leads to larger relative deviations between calculated and experimental results than the calculation of the auto-ignition temperature at constant pressure. In addition to the calculations, a reaction path analysis is performed to explain the observed lowering of the auto-ignition limit of methane/air mixtures by ammonia addition. It is found that this decrease is caused by the formation of NO and NO(2), which enhance the oxidation of methane at low temperatures.


Effects of aging on the digestive solubilization of Cu from sediments.
Solubilization of particulate Cu by different solutions, mimicking digestive fluids of deposit-feeders, was quantified in stable isotope (65)Cu-spiked sediments (with 3 days-2 months Cu-sediment contact time or aging). Copper solubilization generally decreased with prolonged aging. However, such decrease became less evident after 1 month and equilibrium of Cu in sediments could be reached after 2 months. Aging effects on Cu solubilization can be explained by the changes in Cu geochemical fractionation with aging: Cu generally transferred from more mobile phases (carbonate and Fe-Mn associated) to more refractory phases (organic associated and residual phase). Besides Cu geochemical fractionation, digestive fluid composition and different Cu solubilization pathways involved, as well as sedimentary organic content, could all affect the digestive solubilization of Cu and its change with aging. Our results emphasize the necessity of considering Cu aging in laboratory sediment toxicity experiments, and in risk assessment of Cu contaminated sediments.


[Version by internal maneuvers: alternative to cesarean section under precarious conditions. The experience of the University Clinic of Gynecology and Obstetrics].
Obstetrics practised in African University Hospitals is beset with specific constraints. The emergency admission of a woman whose fetus is dead or severely compromised, after an extended labor and an obvious ovular infection, leads to fear an hysterotomy as well as an hysterectomy. The authors specify, under their work conditions, the indications of internal version maneuvers, from 50 cases collected in one year. The fetal prognosis is not taken into account since it concerns dead or very threatened children. As for the maternal prognosis, it is excellent since the average stay is of 48 hours and no severe infection developed, which is not the case for caesarean sections performed under the same circumstances. If a maternal death must be deplored, it concerned a placenta praevia in a distressing conjunction of circumstances since the blood bank was out of blood.


Distal convoluted tubule sexual dimorphism revealed by advanced 3D imaging.
The thiazide-sensitive Na+-Cl- cotransporter (NCC) is more abundant in kidneys of female subjects than of male subjects. Because morphological remodeling of the distal convoluted tubule (DCT) is dependent on NCC activity, it has been generally assumed that there is a corresponding sexual dimorphism in the structure of the DCT, leading to a larger female DCT. Until now, this has never been directly examined. Here, optical clearing techniques were combined with antibody labeling of DCT segment markers, state-of-the-art high-speed volumetric imaging, and analysis tools to visualize and quantify DCT morphology in male and female mice and study the DCT remodeling response to furosemide. We found an unexpected sex difference in the structure of the DCT. Compared with the male mice, female mice had a shorter DCT, a higher cellular density of NCC, and a greater capacity to elongate in response to loop diuretics. Our study revealed a sexual dimorphism of the DCT. Female mice expressed a greater density of NCC transporters in a shorter structure to protect Na+ balance in the face of greater basal distal Na+ delivery yet have a larger reserve and structural remodeling capacity to adapt to unique physiological stresses. These observations provide insight into mechanisms that may drive sex differences in the therapeutic responses to diuretics.


Survey Says: "COVID-19 Lockdown Hits Young Faculty and Clinical Trials".
COVID-19 has severely impacted laboratory research. Analysis of the International Society for Stem Cell Research (ISSCR) member survey has highlighted a particular impact on clinical trials and early-career investigators. The stem cell community needs to support young researchers and ensure that stem cell medicine does not lose its momentum.


A widening longevity gap between people with schizophrenia and general population: A literature review and call for action.
Individuals with schizophrenia have higher mortality rates than the population at large. General mortality rates have declined in developed countries since the early 1970s, extending average lifespan by nearly a decade. This review of eight longitudinal studies of mortality in schizophrenia found that the mean standardized mortality ratio (SMR, a measure of mortality rate in schizophrenia compared to the general population) increased 37%, from 2.2 in the pre-1970s studies to 3.0 in the post-1970s reports. Major changes in societal stigma, healthcare, and economic policy are urgently warranted to ensure that this vulnerable segment of the population participates in the longevity revolution.


In vivo administration of IL-18 can induce IgE production through Th2 cytokine induction and up-regulation of CD40 ligand (CD154) expression on CD4+ T cells.
IL-18 is considered to be a strong cofactor for CD4+ T helper 1 (Th1) cell induction. We have recently reported that IL-18 can induce IL-13 production in both NK cells and T cells in synergy with IL-2 but not IL-12, suggesting IL-18 can induce Th1 and Th2 cytokines when accompanied by the appropriate first signals for T cells. We have now found that IL-18 can act as a cofactor to induce IL-4, IL-10 and IL-13 as well as IFN-gamma production in T cells in the presence of anti-CD3 monoclonal antibodies (mAb). IL-18 can rapidly induce CD40 ligand (CD154) mRNA and surface expression on CD4+ but not CD8+ T cells. The administration of IL-18 alone in vivo significantly increased serum IgE levels in C57BL/6 (B6) and B6 IL-4 knockout mice. Furthermore, the administration of IL-18 plus IL-2 induced approximately 70-fold and 10-fold higher serum levels of IgE and IgG1 than seen in control B6 mice, respectively. IgE and IgG1 induction in B6 mice by administration of IL-18 plus IL-2 was eliminated by the pretreatment of mice with anti-CD4 or anti-CD154, but not anti-CD8 or anti-NK1.1 mAb. These results suggest that IL-18 can induce Th2 cytokines and CD154 expression, and can contribute to CD4+ T cell-dependent, IL-4-independent IgE production.


Verification: model-free phasing with enhanced predicted models in ARCIMBOLDO_SHREDDER.
Structure predictions have matched the accuracy of experimental structures from close homologues, providing suitable models for molecular replacement phasing. Even in predictions that present large differences due to the relative movement of domains or poorly predicted areas, very accurate regions tend to be present. These are suitable for successful fragment-based phasing as implemented in ARCIMBOLDO. The particularities of predicted models are inherently addressed in the new predicted_model mode, rendering preliminary treatment superfluous but also harmless. B-value conversion from predicted LDDT or error estimates, the removal of unstructured polypeptide, hierarchical decomposition of structural units from domains to local folds and systematically probing the model against the experimental data will ensure the optimal use of the model in phasing. Concomitantly, the exhaustive use of models and stereochemistry in phasing, refinement and validation raises the concern of crystallographic model bias and the need to critically establish the information contributed by the experiment. Therefore, in its predicted_model mode ARCIMBOLDO_SHREDDER will first determine whether the input model already constitutes a solution or provides a straightforward solution with Phaser. If not, extracted fragments will be located. If the landscape of solutions reveals numerous, clearly discriminated and consistent probes or if the input model already constitutes a solution, model-free verification will be activated. Expansions with SHELXE will omit the partial solution seeding phases and all traces outside their respective masks will be combined in ALIXE, as far as consistent. This procedure completely eliminates the molecular replacement search model in favour of the inferences derived from this model. In the case of fragments, an incorrect starting hypothesis impedes expansion. The predicted_model mode has been tested in different scenarios.


Does change in hip circumference predict cardiovascular disease and overall mortality in Danish and Swedish women?
Accumulating evidence consistently shows that small hip circumference (HC) is related to increased risk of cardiovascular disease (CVD), coronary heart disease, diabetes, and premature death in women. This study aims to clarify whether this inverse association can be found in both normal- and overweight individuals and if change in HC over time relates to morbidity and mortality risk. HC and 6-year change in HC in relation to the risk for all-cause mortality and CVD morbidity and mortality was investigated in a pooled sample of 2,867 women from the DANISH MONICA study and the Prospective Population Study of Women in Gothenburg with a total of 66,627 person-years of follow-up. Baseline HC was significantly and inversely associated with all-cause and CVD-specific mortality after adjustment for BMI, waist circumference (WC), and other covariates. In stratified analyses, the inverse association was weaker in women with a BMI of more than 25 kg/m2. Six-year change in hip size was not associated with mortality or morbidity endpoints. Our results imply the existence of a basal risk associated with small hip size, which is, however, independent from changes in gluteofemoral body mass and therefore unlikely to be modifiable.


Attitudes and behavior regarding organ donation and transplantation on the part of religious officials in the Eastern Black Sea region of Turkey.
This study evaluated the opinions and attitudes toward organ donation and transplantation on the part of religious officials in the Eastern Black Sea region of Turkey. This descriptive study was performed between December 2008 and November 2009 with 550 among Muslim religious officials, including 541 males and 9 females, who attended educational seminars on the subject arranged in collaboration with Mufti Offices in the provinces of Trabzon, Rize, Gümüşhane, and Giresun in the Eastern Black Sea region of Turkey. A supervised questionnaire administered before the seminars contained questions investigating whether subjects have volunteered to donate organs, reasons for not volunteering, whether they would donate the organs of a close relative in the event such a death, and what they would think in the event they required an organ transplantation themselves. Questions were also asked regarding permission for organ transplantation and donation in 5 main religions. Although no female religious officials had previously volunteered to donate organs, 4 men (0.7%) had done so. After attending seminars, 32 (5.9%) volunteered to donate. The main reasons cited for not volunteering to donate organs were lack of sufficient knowledge of the procedures involved (30.6%) and lack of interest (17.4%). Three hundred five religious officials stated they would not agree to organ donation in the event of the death of a close relative. When asked "What would you think about organ donation and transplantation if you needed an organ transplant now?", 46.0% replied "I would want an organ transplant." Finally, 83.3% of religious officials stated that Islam permits organ donation and transplantation, whereas 13.6% stated that Christianity permits it, other figures being 7.3% for Judaism, 2.5% for Buddhism, and 2.0% for Hinduism. Religious views affect organ donation. The fact that religious officials agreed to donate organs after appropriate education emphasized that Islam is the most sensitive faith on the subject, indicating that the issues can be improved through collaboration with religious officials.


The Association Between Phosphorylated Neurofilament Heavy Chain (pNF-H) and Small Fiber Neuropathy (SFN) in Patients with Impaired Glucose Tolerance.
Small fiber neuropathy (SFN)-the early stage of diabetic peripheral neuropathy (DPN)-progresses gradually and is difficult to diagnose using neurophysiological tests. To facilitate the early diagnosis of SFN, biomarkers for SFN must be identified. The purpose of this study was to investigate the characteristics of SFN in prediabetic patients and the relationship between pNF-H and SFN. 44 IGT patients (inpatients and outpatients) were selected at random. 33 healthy subjects served as controls. Data on clinical characteristics and laboratory parameters were collected. Quantitative sensory testing (QST), electromyography (EMG), and Sudoscan were performed, and pNF-H was measured by ELISA. 24 of the 44 patients with impaired glucose tolerance (IGT) were diagnosed with SFN according to the modified Toronto Diabetic Neuropathy Expert Group consensus criteria. The thermal sensory thresholds of the IGT-SFN group were significantly different from those of the CTRL group (p < 0.05), except for the heat pain threshold. The sensory nerve action potential (SNAP) of the sural nerve was 12.39 in the IGT-SFN group, which was significantly lower than those in the other groups. No significant difference in nerve conduction velocity (NCV) was observed among the three groups. The electrochemical skin conductance (ESC) in the IGT-SFN group was 69.78 ± 14.03uS, which was significantly lower than that in the CTRL group. The pNF-H in the IGT-SFN group was 170.6 (140.0, 223.6) pg/ml, which was significantly higher than those in the CTRL and IGT-non-SFN groups (76.55 and 64.7 pg/ml, respectively). Multivariate regression analysis demonstrated that pNF-H and 2h plasma glucose were independently correlated with SFN; the ORs (95% CI) were 1.429 (1.315, 1.924) and 2.375 (1.157, 4.837), respectively. Serum pNF-H may be associated with SFN in IGT patients, and serum pNF-H could therefore serve as a sensitive biomarker for the detection of SFN.


Membrane-bound globin X protects the cell from reactive oxygen species.
Globin X (GbX) is a member of the globin family that emerged early in the evolution of Metazoa. In vertebrates, GbX is restricted to lampreys, fish, amphibians and some reptiles, and is expressed in neurons. Unlike any other metazoan globin, GbX is N-terminally acylated and anchored in the cell membrane via myristoyl and palmitoyl groups, suggesting a unique function. Here, we compared the capacity of GbX to protect a mouse neuronal cell line from hypoxia and reactive oxygen species (ROS) with that of myoglobin. To evaluate the contribution of membrane-binding, we generated a mutated version of GbX without acyl groups. All three globins enhanced cell viability under hypoxia, with myoglobin having the most pronounced effect. GbX but not myoglobin protected the cells from hydrogen peroxide (H2O2)-induced stress. Membrane-bound GbX was significantly more efficient than its mutated, soluble form. Furthermore, myoglobin and mutated GbX increased production of ROS upon H2O2-treatment, while membrane-bound GbX did not. The results indicate that myoglobin enhances O2 supply while GbX protects the cell membrane from ROS-stress. The ancient origin of GbX suggests that ROS-protection reflects the function of the early globins before they acquired a respiratory role.


Quadratus lumborum block for postoperative analgesia: a systematic review and meta-analysis.
The goal of the present systematic review is to determine the efficacy of the quadratus lumborum block (QLB) in providing postoperative analgesia for abdominal wall and hip surgeries when compared with placebo or other analgesic techniques. Electronic databases (Medline, Embase, Cochrane Central, and Scopus) were searched for keywords and controlled vocabulary terms related to QLB from their inception to November 2019. The included studies compared ultrasound-guided single-injection QLB to placebo and other analgesic techniques in adult patients. Forty-two randomized-controlled trials provided the data for this systematic review. Eight studies were assessed as high risk of bias in at least one domain. The included studies had significant heterogeneity with regard to the type of surgery, comparator groups, and outcomes measured; therefore, a limited quantitative analysis was undertaken for the comparison of QLB vs no block or placebo in patients undergoing Cesarean delivery only. For Cesarean delivery, the QLB reduced the opioid use by 24.1 (95% confidence interval, 17.3 to 30.9) mg oral morphine equivalents in the first postoperative 24 hr compared with no block or placebo with no difference in pain scores at rest. For other surgical procedures, the pain scores and opioid use were lower in the QLB group when compared with placebo or no regional anesthesia technique. When compared with other regional anesthetic techniques, the analgesic benefit of QLB was marginal. Quadratus lumborum block provided analgesic benefits compared with placebo for use in the abdominal wall and hip surgery, with only marginal benefits compared with other regional analgesic techniques. The identified studies used different variants of QLB in many different surgery types. These findings and conclusions, therefore, should be considered preliminary. PROSPERO (CRD42018095965); registered 6 June 2018.


Role of electrocardiography in identifying right ventricular dysfunction in acute pulmonary embolism.
The role of electrocardiography in identifying right ventricular (RV) dysfunction in acute pulmonary embolism (APE) was evaluated in 81 patients with APE. The electrocardiographic markers studied were T-wave inversion in leads V1 to V3, the S1Q3T3 pattern, right bundle branch block, and sinus tachycardia. T-wave inversion in leads V1 to V3 had the greatest sensitivity and diagnostic accuracy for identifying RV dysfunction in patients with APE. The S1Q3T3 pattern and right bundle branch block had good specificity but moderate accuracy.


Gravity-efficient motor control is associated with contraction-dependent intracortical inhibition.
In humans, moving efficiently along the gravity axis requires shifts in muscular contraction modes. Raising the arm up involves shortening contractions of arm flexors, whereas the reverse movement can rely on lengthening contractions with the help of gravity. Although this control mode is universal, the neuromuscular mechanisms that drive gravity-oriented movements remain unknown. Here, we designed neurophysiological experiments that aimed to track the modulations of cortical, spinal, and muscular outputs of arm flexors during vertical movements with specific kinematics (i.e., optimal motor commands). We report a specific drop of corticospinal excitability during lengthening versus shortening contractions, with an increase of intracortical inhibition and no change in spinal motoneuron responsiveness. We discuss these contraction-dependent modulations of the supraspinal motor output in the light of feedforward mechanisms that may support gravity-tuned motor control. Generally, these results shed a new perspective on the neural policy that optimizes movement control along the gravity axis.


Folding and stability of the three-stranded beta-sheet peptide Betanova: insights from molecular dynamics simulations.
The dynamics of the three-stranded beta-sheet peptide Betanova has been studied at four different temperatures (280, 300, 350, and 450 K by molecular dynamics simulation techniques, in explicit water. Two 20-ns simulations at 280 K indicate that the peptide remains very flexible under "folding" conditions sampling a range of conformations that together satisfy the nuclear magnetic resonance (NMR)-derived experimental constraints. Two simulations at 300 K (above the experimental folding temperature) of 20 ns each show partial formation of "native"-like structure, which also satisfies most of the NOE constraints at 280 K. At higher temperature, the presence of compact states, in which a series of hydrophobic contacts remain present, are observed. This is consistent with experimental observations regarding the role of hydrophobic contacts in determining the peptide's stability and in initiating the formation of turns and loops. A set of different structures is shown to satisfy NMR-derived distance restraints and a possible mechanism for the folding of the peptide into the NMR-determined structure is proposed.


The Italian law on informed consent and advance directives: New rules of conduct for the autonomy of doctors and patients in end-of-life care.
Italy has long lacked a law regulating patients' informed consent and advance directives (ADs). All previous attempts to introduce a law on this matter failed to reach positive outcomes, and aroused heated ideological debate over the exact meaning of life and death. We report on the new law on informed consent and ADs approved by the Italian Parliament on 14th December 2017. We analyse the new law and discuss the main ethical points connected with it, in the Italian context and in comparison with the international situation. The law provides for fundamental ethical principles and important guidelines: respect for patients' self-determination in all phases of life, option to refuse or interrupt life-sustaining treatments, including artificial nutrition and hydration, the legitimacy of end-of-life decisions, and the implementation of palliative care to ease suffering and pain. The effects of the new law must be tested in the field. Its objectives will be achieved if, in clinical practice, ADs are able to satisfactorily represent informed personal preferences through patients' relationships with their physicians, as part of personalized advance care planning. Future studies are necessary to assess the impact of the new law in Italy.


[Effect of nembutal anesthesia, electric stimulation of the paws and elaboration of a defensive reflex on acetylcholinesterase activity and protein content in different regions of rat brain].
Pentobarbitone sodium anaesthesia was found to produce an increase in protein content in some regions of the rat brain, i. e. posterior cortex, caudate nucleus, and a decrease in protein content in the ventral cortex. Acetylcholinesterase expressed in terms of wet weight was found to increase in the cerebellum, medulla, and to decrease in the medial cortex, hippocampus, thalamus and caudate nucleus. The changes in activity were not explicable in terms of a direct effect of the anaesthetic on the enzyme. A decrease in protein content of rat brain was observed in the frontal cortex, ventral cortex, hippocampus and caudate nucleus after electrical shocks. Following shock avoidance conditioning procedure (shuttle-box), decreases in protein content were observed in the medial cortex, posterior cortex, cerebellum and ventral cortex; in the thalamus an increase in protein content was observed. Changes in AChE activity were observed following footshock in the frontal cortex and medulla where there was an increase in activity and in the caudate nucleus, hypothalamus, thalamus, and olfactory tubercule where there was a decrease in activity. Following shock avoidance conditioning the activity of the AChE increased in posterior cortex, hippocampus, thalamus and hypothalamus and the activity of the enzyme decreased in the ventral cortex.


Phase I evaluation of peplomycin with special reference to pulmonary toxicity.
In the search for bleomycin analogues with less pulmonary toxicity than bleomycin itself, peplomycin was selected for a phase I clinical trial, based on experimental animal data. Eighteen patients received peplomycin at three exploratory levels. Six patients were treated at a level of 5 mg/m2, 8 patients at 10 mg/m2 and 4 patients at 15 mg/m2 of peplomycin, each dosage being given twice weekly intravenously. Pulmonary function tests were performed prior to treatment and serially thereafter. Pulmonary toxicity was encountered when the administered total dose of peplomycin was in the range 190-350 mg in patients who had received either 10 or 15 mg/m2 twice weekly. Pulmonary toxicity was not observed when the dosage of peplomycin was restricted to 5 mg/m2 twice weekly. During the trial no haematological, hepatic or renal changes induced by the drug were observed. Skin changes, stomatitis and fever were observed with increasing frequency the higher the cumulative dose of peplomycin, and these effects were similar to those seen with bleomycin. Two of fifteen patients with cervical cancer obtained a partial response, lasting 1 and 2 months respectively. Although peplomycin is free from pulmonary toxicity at a dose of 5 mg/m2 twice weekly, the maximum tolerated cumulative dose has still to be defined.


Gold nanoparticle-adjuvanted S protein induces a strong antigen-specific IgG response against severe acute respiratory syndrome-related coronavirus infection, but fails to induce protective antibodies and limit eosinophilic infiltration in lungs.
The spike (S) protein of coronavirus, which binds to cellular receptors and mediates membrane fusion for cell entry, is a candidate vaccine target for blocking coronavirus infection. However, some animal studies have suggested that inadequate immunization against severe acute respiratory syndrome coronavirus (SARS-CoV) induces a lung eosinophilic immunopathology upon infection. The present study evaluated two kinds of vaccine adjuvants for use with recombinant S protein: gold nanoparticles (AuNPs), which are expected to function as both an antigen carrier and an adjuvant in immunization; and Toll-like receptor (TLR) agonists, which have previously been shown to be an effective adjuvant in an ultraviolet-inactivated SARS-CoV vaccine. All the mice immunized with more than 0.5 µg S protein without adjuvant escaped from SARS after infection with mouse-adapted SARS-CoV; however, eosinophilic infiltrations were observed in the lungs of almost all the immunized mice. The AuNP-adjuvanted protein induced a strong IgG response but failed to improve vaccine efficacy or to reduce eosinophilic infiltration because of highly allergic inflammatory responses. Whereas similar virus titers were observed in the control animals and the animals immunized with S protein with or without AuNPs, Type 1 interferon and pro-inflammatory responses were moderate in the mice treated with S protein with and without AuNPs. On the other hand, the TLR agonist-adjuvanted vaccine induced highly protective antibodies without eosinophilic infiltrations, as well as Th1/17 cytokine responses. The findings of this study will support the development of vaccines against severe pneumonia-associated coronaviruses.


Robotic-arm assisted total knee arthroplasty has a learning curve of 16 cases and increased operative time of 12 min.
Robotic-arm assisted systems are increasingly used for knee arthroplasty, however introduction of new systems can involve a learning curve. We aimed to define the learning curve in terms of operative time and component placement/sizing of a robotic system for total knee arthroplasty (TKA) in a team of experienced surgeons, and to investigate mid-term patient outcomes. A total of 101 consecutive patients underwent primary robotic-arm assisted TKA by three surgeons (mean 2 year follow-up). Operative times, component placement, implant sizing and reoperations were recorded. Cumulative Summation (CUSUM) was used to analyse learning curves. Patient outcomes were compared between learning and proficiency phases. The learning curve was 16 cases, with a 12-min increase in operative time (P < 0.01). Once proficiency was achieved, the greatest time reductions were seen for navigation registration (P = 0.003) and bone preparation (P < 0.0001). A learning curve was found with polyethylene (PE) insert sizing (P = 0.01). No differences were found between learning and proficiency groups in terms of implant survival (100% and 97%, respectively, NS) or patient-reported outcome measures at 2 years (NS). Introduction of a robotic-arm assisted system for TKA led to increased operative times for navigation registration and bone preparation, and a learning curve with PE insert sizing. No difference in patient outcomes between learning and proficiency groups at 2 years was found. These findings can inform surgeons' expectations when starting to use robotic-assisted systems.


3,5-dibenzoyl-1,4-dihydropyridines: synthesis and MDR reversal in tumor cells.
Fifteen 4-phenyl-3,5-dibenzoyl-1,4-dihydropyridines (BzDHPs) (1-15) substituted at the 4-phenyl ring were synthesized and compared to their cytotoxic activity and multidrug resistance (MDR)-reversing activity in in vitro assay systems. Among them, 2-CF3 (5) (IC50=8.7 microM), 2-Cl (11) (IC50=7.0 microM) and 3-Cl (12) (IC50=7.0 microM) derivatives showed the highest cytotoxic activity against human oral squamous carcinoma (HSC-2) cells. The activity of P-glycoprotein (Pgp) response for MDR in tumor cells was reduced by some of derivatives (3, 4, 8, 12), verapamil (VP) and nifedipine (NP). These data suggest that 3,5-dibenzoyl-4-(3-chlorophenyl)-1,4-dihydro-2,6-dimethylpyridine (12) can be recommended as a new drug candidate for MDR cancer treatment.


Selective phosphodiesterase inhibitors for the treatment of asthma and chronic obstructive pulmonary disease.
Phosphodiesterase type 4 plays an important role in modulating the activity of cells that are involved in the inflammatory processes occurring in chronic obstructive pulmonary disease and asthma. During the past decade, interest has focused on the development of selective inhibitors of phosphodiesterase-4, in an attempt to generate new treatment modalities that control airway inflammation not only in patients with asthma but also in those with chronic obstructive pulmonary disease. The aim of the present review is to discuss the results of recent clinical intervention studies with phosphodiesterase-4 inhibitors in patients with asthma or chronic obstructive pulmonary disease.


Modulation of Phase Separation by RNA: A Glimpse on N6-Methyladenosine Modification.
Phase separation is the driving force behind formation of various biomolecular condensates (BioMCs), which sub-compartmentalize certain cellular components in a membraneless manner to orchestrate numerous biological processes. Many BioMCs are composed of proteins and RNAs. While the features and functions of proteins are well studied, less attention was paid to the other essential component RNAs. Here, we describe how RNA contributes to the biogenesis, dissolution, and properties of BioMCs as a multivalence providing scaffold for proteins/RNA to undergo phase separation. Specifically, we focus on N6-methyladenosine (m6A), the most widely distributed dynamic post-transcriptional modification, which would change the charge, conformation, and RNA-binding protein (RBP) anchoring of modified RNA. m6A RNA-modulated phase separation is a new perspective to illustrate m6A-mediated various biological processes. We summarize m6A main functions as "beacon" to recruit reader proteins and "structural switcher" to alter RNA-protein and RNA-RNA interactions to modulate phase separation and regulate the related biological processes.