Fractional anisotropy in the centrum semiovale as a quantitative indicator of cerebral white matter damage in the subacute phase in patients with carbon monoxide poisoning: correlation with the concentration of myelin basic protein in cerebrospinal fluid.
Carbon monoxide (CO) poisoning leads to demyelination of cerebral white matter (CWM) fibers, causing chronic neuropsychiatric symptoms. To clarify whether fractional anisotropy (FA) from diffusion tensor imaging in the centrum semiovale can depict demyelination in the CWM during the subacute phase after CO inhalation, we examined correlations between FA in the centrum semiovale and myelin basic protein (MBP) in cerebrospinal fluid. Subjects comprised 26 adult CO-poisoned patients ≤60 years old. MBP concentration was examined for all patients at 2 weeks after CO inhalation. The mean FA of the centrum semiovale bilaterally at 2 weeks was also examined for all patients and 21 age-matched healthy volunteers as controls. After these examinations, the presence of chronic symptoms was checked at 6 weeks after CO poisoning. Seven patients displayed chronic symptoms, of whom six showed abnormal MBP concentrations. The remaining 19 patients presented no chronic symptoms and no abnormal MBP concentrations, with MBP concentrations undetectable in 16 patients. The MBP concentration differed significantly between patients with and without chronic symptoms. The mean FA was significantly lower in patients displaying chronic symptoms than in either patients without chronic symptoms or controls. After excluding the 16 patients with undetectable MBP concentrations, a significant correlation was identified between MBP concentration and FA in ten patients. The present results suggest that FA in the centrum semiovale offers a quantitative indicator of the extent of demyelination in damaged CWM during the subacute phase in CO-poisoned patients.


Factors associated with malaria infection in Mudzi District, Mashonaland East Zimbabwe, 2019: a case-control study.
Kondo Rural Health Centre recorded 27 malaria patients between the 27th of January 2019 and the 2nd of February 2019 against an epidemic threshold of 19 with the malaria outbreak being confirmed on the 5th of February 2019. Indoor residual spraying as part of integrated vector management control activities had been done in the district before the onset of the rainy season as well as social behaviour change communication but residents were contracting malaria. We, therefore, investigated the risk factors associated with this outbreak to recommend scientifically effective prevention and control measures. We conducted a 1:1 unmatched case-control study. A case was a resident of Mudzi from the 4th of February 2019 who had a positive rapid diagnostic test for malaria randomly selected from the clinic's line list whilst controls were randomly selected from the neighbourhood of cases. Pretested interviewer-administered questionnaires were used to collect information on demographic characteristics, knowledge and practices of residents in malaria prevention. Data were analysed using Epi info 7. A total of 567 confirmed malaria cases was recorded with an overall attack rate of 71.7 per 1000 population. Sixty-three case-control pairs were interviewed. The majority of cases 78% (49/63) were from Makaza, Chanetsa and Nyarongo villages which are within 3 km from Vhombodzi dam. A stagnant water body near a house [aOR = 8.0, 95%CI = (2.3-28.6)], engaging in outdoor activities before dawn or after dusk [aOR = 8.3, 95%CI = (1.1-62.7)] and having a house with open eaves [aOR = 5.4, 95%CI = (1.2-23.3)] were independent risk factors associated with contracting malaria. Wearing long-sleeved clothes when outdoors at night [aOR = 0.2, 95%CI = (0.1-0.4)] was protective. A stagnant water pool close to the homestead and engaging in outdoor activities before dawn and after dusk were modifiable risk factors associated with the malaria outbreak despite the community being knowledgeable on the transmission and prevention of malaria. Community sensitisation and mobilisation in the destruction of stagnant water bodies and cutting of tall grass around homesteads were recommended measures to contain the outbreak.


Unusual lung tumours: an illustrated review of CT features suggestive of this diagnosis.
This review presents an imaging-centered approach to the diagnostic challenge of uncommon lung tumours in adults. Emphasis is placed on features that may be used to differentiate these tumours including fat content, tumour site, multifocality, calcification and predominant pattern of involvement.


Development of experimental treatments for patients with retinitis pigmentosa.
Retinitis pigmentosa (RP) is a group of inherited diseases that lead to degeneration of the retina and decreased vision. The World Health Organization reports around 1,300 million people affected by some type of visual impairment worldwide. The prevalence is 1 in every 4000 inhabitants and it is the first cause of blindness of genetic origin, frequent in men with a percentage of 60% and 40% in women. There is a lack of information on this pathology in the world, mainly on the existing treatments for this disease, so this bibliographic review aims to update the existing or under-study treatments and inform the limitations of each of these therapies. This review of scientific literature was carried out by consulting databases such as PubMed and Web of science, the search will be limited to articles from the years 2018-2022. There are several types of therapy in studies: gene therapy, transcorneal electrical stimulation, use of neuroprotectors, optogenic therapy, stem cell transplants and oligonucleotide therapy, which will be discussed in this article, both their benefits and the existing barriers in each treatment experimental. In conclusion, each of these therapies promises a viable treatment in the future for selective groups of people with retinitis pigmentosa, however, some therapies have shown benefit at the beginning of the disease, losing their efficacy in the long term.


Biofuels from microalgae.
Microalgae are a diverse group of prokaryotic and eukaryotic photosynthetic microorganisms that grow rapidly due to their simple structure. They can potentially be employed for the production of biofuels in an economically effective and environmentally sustainable manner. Microalgae have been investigated for the production of a number of different biofuels including biodiesel, bio-oil, bio-syngas, and bio-hydrogen. The production of these biofuels can be coupled with flue gas CO2 mitigation, wastewater treatment, and the production of high-value chemicals. Microalgal farming can also be carried out with seawater using marine microalgal species as the producers. Developments in microalgal cultivation and downstream processing (e.g., harvesting, drying, and thermochemical processing) are expected to further enhance the cost-effectiveness of the biofuel from microalgae strategy.


A comparative study of biofilm removal with hand, rotary nickel-titanium, and self-adjusting file instrumentation using a novel in vitro biofilm model.
This study sought to present a standardized biofilm model in extracted teeth with an artificial apical groove to quantify the efficacy of hand, rotary nickel-titanium, and self-adjusting file (SAF) instrumentation in biofilm bacteria removal. Thirty-six extracted single-rooted teeth with oblong canals were selected. Each tooth was split longitudinally, and a 0.2-mm-wide groove was placed in the apical 2 to 5 mm of the canal. After growing mixed bacteria biofilm inside the canal under an anaerobic condition, the split halves were reassembled in a custom block, creating an apical vapor lock. Teeth were randomly divided into 3 treatment groups (n = 10 per group) using the K-file, ProFile (Dentsply Tulsa Dental Products, Tulsa, OK), and the SAF (ReDent-Nova, Ra'anana, Israel). Irrigation consisted of 10 mL 3% NaOCl and 4 mL 17% EDTA. Six teeth received no treatment. Areas inside and outside the groove were examined using a scanning electron microscope. The scanning electron microscope showed a consistently thick layer of biofilm grown in the canals of the control group after 4 weeks. Within the groove, a smaller area remained occupied by bacteria after the use of the SAF compared with the ProFile and the K-file (3.25%, 19.25%, and 26.98%, respectively; P < .05). For all groups, significantly more bacteria were removed outside the groove than inside (P < .05). No statistical differences were found outside the groove (P > .05). Although all techniques equally removed bacteria outside the groove, the SAF reduced significantly more bacteria within the apical groove. No technique was able to remove all bacteria. This biofilm model represents a potentially useful tool for the future study of root canal disinfection.


Energy eigenvalue surfaces close to a degeneracy of unbound states: crossings and anticrossings of energies and widths.
The rich phenomenology of crossings and anticrossings of energies and widths, as well as the sudden change in shape of the S(E)-matrix pole trajectories, observed in an isolated doublet of resonances when one control parameter is varied, is fully explained in terms of sections of the energy eigenvalue surfaces in parameter space close to a degeneracy point.


[Formula: see text]Is there an effective dose of aerobic exercise associated with better executive function in youth with attention deficit hyperactivity disorder?
Attention-deficit/hyperactivity disorder (ADHD) is the most prevalent neurodevelopmental disorder in children, and it's linked to a higher risk of academic failure, interpersonal issues, mental illness, and criminality. Despite several of the comparative and detailed reviews on the effects of ADHD interventions on core symptoms, there is no data summarizing the effects of aerobic exercise (AE) on executive functions (EFs). Therefore, this study aimed to systematically review and determine the relationship between AE (acute and chronic) dosage and EFs (attention, inhibition, set-shifting, and working memory) in youth with ADHD. The consideration of how AE dosage impacts aspects of EFs has not been investigated in detail previously. The study adhered to PRISMA guideline. Sixdatabases were searched without any date restrictions, up to February 2021, for articles relating to AE interventions to influence EFs in youth with ADHD≤18 years old. Quality assessment of the reviewed papers was addressed. Thirteen studies met the inclusion criteria. Improvements in all aspects of EFs were reported after children with ADHD engaged in acute AE lasting 20-30 minutes with at least moderate intensity (65% ≤ 80% HRmax).Furthermore, chronic exercise that lasts at least 45 minutes and in the range of moderate tohigh intensity (i.e., 60% ≤ 75% HRmax), 3 days/week for 6-12, elicits additional benefits in inhibition and set-shifting.Different dosage of AE might differently influence aspects of EFs; however, this finding rests on preliminary evidence at this stage and thus should be treated with caution.It is necessary to establish suitable interventions with regard to the dosage of AE types to improve EFs in young people with ADHD.


Analysis of follicular events in owl monkeys (Aotus azarai infulatus) using B-mode and Doppler ultrasound.
Ultrasound (B-mode) was used to analyze follicular events in 12 trained female owl monkeys (Aotus azarai infulatus). The animals were examined every 48 hours for over 90 days to measure and map follicular growth in both ovaries and to measure (using Doppler velocimetry) local hemodynamic changes during the peri-ovulatory stage. There were 44 follicular growth events, each with two or three follicular waves, and a mean ± SEM interval between events of 17 ± 1.13 days. There were various hemodynamic changes during follicular growth; both vascular resistance index and pulsatility index decreased during the time when the follicle diameter peaked. Thus, both B-mode and Doppler ultrasound were useful for monitoring ovarian follicular events in owl monkeys.


Characterization of neuropeptide Y, Y(2) receptor knockout mice in two animal models of learning and memory processing.
Neuropeptide Y (NPY) and, in particular, the Y2 receptor subtype, has been suggested to be involved in learning and memory processing. However, the precise role of Y2 receptors in learning and memory remains unclear. In the present study, mice lacking NPY Y2-type receptors were assessed in two animal models of learning and memory processing. We found that NPY Y2-/- mice displayed a deficit on the probe trial in the Morris water maze task, whereas acquisition performance, swim speed, and visible platform performance did not differ significantly between groups. In addition, NPY Y2-/- mice exhibited a marked deterioration in object memory 6 h, but not 1 h, following initial exposure in the object recognition test. Both groups of mice showed similar locomotor activity profiles in a low-stress, open field test. These data support the hypothesis that Y2 receptors are involved in the regulation of learning and memory processing.


Chronic kidney disease is associated with neovascularization and intraplaque hemorrhage in coronary atherosclerosis in elders: results from the Hisayama Study.
There is little information regarding whether patients with chronic kidney disease (CKD) have a high incidence of vulnerable plaques in their coronary arteries. To gain additional evidence on this, we conducted a population-based study by randomly selecting 126 subjects from 844 consecutive autopsies of elderly residents of Hisayama, Japan. We then determined the relationships of CKD with neovascularization and intraplaque hemorrhage in coronary atherosclerosis with the subjects classified into four categories based on their estimated glomerular filtration rate (eGFR). Areas of oxidized low-density lipoprotein (oxLDL) and vascular endothelial growth factor (VEGF) expression, assessed by immunohistochemistry in a total of 375 coronary arteries, increased significantly with decreasing eGFR. A lower eGFR was also associated with increased numbers of newly formed blood vessels. These relationships remained substantially unchanged after adjustment for confounding factors. The multivariate-adjusted odds ratio of the presence of intraplaque hemorrhages was 6.2 (95% confidence interval, 1.1-35.0) in patients with an eGFR <30 ml/min/1.73 m(2) compared with those with an eGFR of ≥ 60 ml/min/1.73 m(2). Thus, elderly patients with CKD have intimal neoangiogenesis and an increased risk of intraplaque hemorrhage in coronary arteries, possibly favored by local accumulation of oxLDL and VEGF.


Label-free detection of single-stranded DNA binding protein based on a cantilever array.
We report a simple and sensitive method for label-free detection of single-stranded DNA-binding protein (SSBP) based on an array of microfabricated cantilevers. The single-stranded DNA (ssDNA) was immobilized on the surface of the sensing cantilevers to detect SSBP, while the reference cantilevers were modified with 6-mercapto-1-hexanol to detect any unwanted cantilever deflection. The differential deflection signals that reveal specific SSBP-ssDNA binding have been found to depend on the SSBP concentration. Using the cantilever array sensor we can detect SSBP in the concentration range from 0.01 to 7 μg mL(-1). Other proteins, such as thrombin or bovine serum albumin induced no significant deflection of the cantilevers. Our results show the potential for the application of cantilever array sensor system as a powerful tool to detect proteins with high sensitivity and specificity.


Solubility fo fibrin clots in monochloroacetic acid. A reflection of serum pepsinogen levels.
Fibrin clots formed from normal plasma dissolve readily in 1% monochloroacetic acid at 37 C. However, if the clots are washed thoroughly before the acid is added, they are no longer soluble. The agent present in the serum which causes dissolution of the fibrin clot was isolated and identified as pepsinogen. Because of the low pH of monochloroacetic acid the pepsinogen is activated and the clots are digested, simulating the dispersion of a fibrin clot which occurs in the absence of fibrin-stabilizing factor (factor XIII). Because of its higher pH, urea will not activate pepsinogen and is therefore a better agent to screen factor XIII deficiencies.


Transcriptional activation of the heme oxygenase gene by heme and cadmium in mouse hepatoma cells.
Treatment of mouse hepatoma (Hepa) cells with heme or cadmium chloride in serum-free medium causes a rapid increase in the steady-state level of heme oxygenase (HO) messenger RNA. This increase is both dose- and time-dependent. Maximum accumulation of HO mRNA is observed 3 h after addition of either agent. Treatment of Hepa cells with heme or CdCl2 also stimulates the transcription of the HO gene, as judged by in vitro nuclear transcription run-on assays. The maximum rate of HO gene transcription occurs 2 h after treatment with either agent. Comparison of the relative increase in the rate of HO gene transcription with the relative increase in the level of HO mRNA demonstrates that transcriptional activation is the primary mechanism by which heme and cadmium produce the accumulation of HO mRNA in Hepa cells. Cadmium may also influence other processes involved in the expression of HO, since the time course of mRNA accumulation diverges from that of gene transcription. However, neither heme nor cadmium alters the rate of HO mRNA degradation. Cobalt chloride and heat shock, which are potent inducers of HO mRNA in rat liver and rat C6 glioma cells, respectively, have only a small effect on the level of HO mRNA in mouse hepatoma cells.


Development of a spatially transparent electrotactile display and its performance in grip force control.
An important function for a tactile navigation system of a handheld tool, such as a surgical scalpel, is the spatial transparency of the device. This paper proposed a new tactile display that can augment touch sensation at the finger pulps without the need for a stimulator between the tool and the finger pulps. We utilized transcutaneous electrical nerve stimulation at the middle phalanx of a finger to separate the stimulated and the perceived areas. In order to verify the effects of the spatial transparency, the performances of grip force control were examined. The results indicated that the proposed display was effective in helping the user to maintain the stable control of the grip force when using a handheld tool.


Evaluation of the effectiveness of a novel oral formulation of sarolaner (Simparica™) for the treatment and control of fleas on dogs.
The efficacy of a single oral dose of a novel isoxazoline, sarolaner (Simparica™, Zoetis), for the treatment and control of flea infestations on dogs was confirmed in five laboratory studies. The studies were conducted using adult purpose-bred Beagles and/or mixed breed dogs. All animals were individually identified and housed, and were allocated randomly to treatment with either placebo or sarolaner (eight to 10 per group) based on pretreatment parasite counts. Three studies used cat flea (Ctenocephalides felis felis) strains recently isolated from the field from the US, EU, or Australia; in the fourth study a laboratory strain (KS1) with documented tolerance to a number of insecticides such as fipronil, imidacloprid, and permethrin was used. In the fifth study, dogs were infested with dog fleas, Ctenocephalides canis. Dogs were treated orally on Day 0 with a placebo or a sarolaner tablet providing a minimum dose of 2mg/kg. Dogs were infested with approximately 100 unfed, adult fleas prior to treatment and at weekly intervals post-treatment. Comb counts were conducted to determine the numbers of viable fleas at 24h after treatment and after each subsequent infestation. Efficacy against C. felis and C. canis was 99.8-100% from treatment through Day 35. In all five studies, elimination of existing infestations was achieved within 24h after dosing, with only a single live C. felis found on one dog on Day 1. Similarly, control of flea challenges was achieved within 24h after infestation throughout the 35day study periods, with only single live C. felis found on two dogs on Day 28 in one study, and on a single dog on Day 35 in another study. There were no adverse reactions to treatment with sarolaner. These studies confirmed that a single oral dose of sarolaner at 2mg/kg provided highly effective treatment of existing C. felis infestations and persistent control of C. felis on dogs for 35days after treatment. Efficacy equivalent to that seen with C. felis was confirmed against C. canis and a known insecticide-tolerant strain of C. felis.


Modeling stoichiometric and oxygen defective TiO2 anatase bulk and (101) surface: structural and electronic properties from hybrid DFT.
We present a periodic hybrid DFT investigation of the structural and electronic properties of both stoichiometric and oxygen-defective TiO2 anatase bulk and (101) surface, in singlet and triplet spin states. In all cases, an excellent agreement with available photoelectron spectroscopy data has been obtained, reproducing the offsets of the deep defect levels positions from the conduction band minimum of TiO2 created upon oxygen vacancy (VO) formation. For the bulk, different local structural polaronic distortions around the VO site have been evidenced depending on the spin state considered. Although a similar conclusion has been drawn for the defective surface for the nine different vacancy positions which have been considered, large migration of the twofold coordinated surface O atom has also been evidenced, up to the initial vacancy site in some cases. The very good agreement obtained with available experimental data regarding the offsets from the conduction band minimum of the deep defect levels positions both for the bulk and the (101) surface of TiO2 anatase is encouraging for the application of the proposed hybrid-based computational strategy to TiO2 surface-related processes such as TiO2-based photocatalysis in which oxygen vacancies are known to play a key role. All calculations have been performed with Crystal17, considering different hybrid functionals with both effective core pseudopotentials and all-electron atom-centered basis sets, as well as additional empirical dispersion effects with the D2 and D3 models.


Socioeconomic position, maternal IQ, home environment, and cognitive development.
To assess whether socioeconomic position, maternal intelligence (IQ), and the home environment are inter-related to cognitive development in childhood. Prospective cohort study (n = 723) with cognitive tests at ages 2, 4, 7, and 11 to 13 years. There were statistically significant positive associations of father's occupational prestige, Home Observation for Measurement of Environment (HOME) score, and maternal IQ with cognitive performance in childhood. After adjustment for confounding factors, there was an increase in cognitive development by 0.8 to 2.0, 2.9 to 4.8, and 4.2 to 9.0 points for a 10-unit increment in father's occupational prestige, maternal IQ, and HOME score, respectively. These results demonstrate that socioeconomic position, maternal IQ, and the home environment are independently and positively predictive of children's cognitive development. These findings provide additional rationale for implementing social policies that reduce socioeconomic inequalities.


Incomplete distal denervation and recurrent ulcer after parietal cell vagotomy.
Twenty-five patients with chronic duodenal ulcer and subjected to parietal cell vagotomy were evaluated as either underdenervated or optimally denervated, depending on whether parietal cells were present distal to the last branch of Latarjet's nerve. A 5-year follow-up study demonstrated recurrent ulcer in three of five judged as underdenervated. In addition to the possible technical error of distal and proximal dissection, the importance of the inborn error of incomplete distal denervation is confirmed.


Childhood Executive Function Predicts Internalizing and Externalizing Symptoms in Emerging Adults With and Without Autism: A 10-Year Longitudinal Study.
Individuals with autism spectrum disorder (ASD) and typically developing individuals were assessed on three neuropsychological tests of executive function (EF) and on scales of autism symptoms and co-occurring internalizing and externalizing symptoms at baseline (T1; N = 88, Mage = 11.8 years, 73% males), 2-year (T2; 99% retention, Mage = 13.9 years), and 10-year follow-ups (T3; 75% retention, Mage = 21.4 years). An EF composite score from T1 significantly predicted internalizing symptoms at T2 (β = .228) and internalizing and externalizing symptoms at T3 (β = .431 and .478, respectively), when controlling for age and autism symptoms. OThe findings suggest that EF difficulties are a long-term risk factor for more co-occurring symptoms. .


Monitoring the organization and dynamics of bovine hippocampal membranes utilizing Laurdan generalized polarization.
Organization and dynamics of cellular membranes in the nervous system are crucial for the function of neuronal membrane receptors. The lipid composition of neuronal cells is unique and has been correlated with the increased complexity in the organization of the nervous system during evolution. Previous work from our laboratory has established bovine hippocampal membranes as a convenient natural source for studying neuronal receptors such as the G-protein coupled serotonin1A receptor. In this paper, we have explored the organization and dynamics of bovine hippocampal membranes using the amphiphilic environment-sensitive fluorescent probe Laurdan. Our results show that the emission spectra of Laurdan display an additional red shifted peak as a function of increasing temperature in native as well as cholesterol-depleted membranes and liposomes made from lipid extracts of the native membrane. Interestingly, wavelength dependence of Laurdan generalized polarization (GP) in native membranes indicates the presence of an ordered gel-like phase at low temperatures, whereas characteristics of the liquid-ordered phase are observed at high temperatures. Similar experiments performed using cholesterol-depleted membranes show fluidization of the membrane with increasing cholesterol depletion. In addition, results from fluorescence polarization of DPH indicate that the hippocampal membrane is fairly ordered even at physiological temperature. The temperature dependence of Laurdan excitation GP provides a measure of the apparent thermal transition temperature and extent of cooperativity in these membranes. Analysis of time-resolved fluorescence measurements of Laurdan shows reduction in mean fluorescence lifetime with increasing temperature due to change in environmental polarity. These results constitute novel information on the dynamics of hippocampal membranes and its modulation by cholesterol depletion monitored using Laurdan fluorescence.


Evaluation: methods and strategy for evaluation--Oregon.
The authors describe the development of comprehensive mechanisms for the evaluation of Oregon's newly funded tobacco control efforts through building on established surveillance systems.


Expression regulation of MAO isoforms in monocytic cells in response to Th2 cytokines.
Th2-cytokines, such as interleukins-4 and -13 (IL-4, IL-13), have been identified as alternative stimuli of monocytes/macrophages. We have recently profiled the gene-expression pattern of IL-4-treated human peripheral monocytes and found that 15-lipoxygenase-1 (15-LOX1) and monoamine oxidase A (MAO-A) are among the five most strongly upregulated gene products in IL-4-treated cells. Transfection of monocytic cells (U937) with 15-LOX1 also induced MAO-A expression. These data suggested that 15-LOX1 products might play a role in the IL4-induced signaling cascade leading to expression of MAO-A in human monocytes. To test this hypothesis we incubated wild-type and 15-LOX1-transfected U937 cells with different concentrations of either IL-4 or 15-LOX-products [13S-H(p)ODE, 15S-H(p)ETE] and quantified the expression of 15-LOX1, MAO-A, and MAO-B by activity assays and real-time RT-PCR. Wild-type U937 cells express neither MAO-A nor MAO-B, but after three days of IL4 treatment, MAO-A mRNA was detected. A similar isoform-specific expression of MAO-A mRNA was observed when U937 cells were transfected with 15-LOX1 or when the cells were incubated with primary 15-LOX1 products (hydroperoxy fatty acids) or H2O2. In contrast, the corresponding hydroxy fatty acids were ineffective. These data indicate that increased intracellular peroxide concentrations (oxidative stress) induce MAO-A expression in monocytes/macrophages, which normally do not express the enzyme. Our findings also suggest that IL-4-induced upregulation of MAO-A expression in human peripheral monocytes may proceed via 15-LOX1-dependent and 15-LOX1-independent pathways. The biological role of MAO-A expression for monocyte function is discussed.


Trypanosome hydrolases and the blood-brain barrier.
African trypanosomes cross the blood-brain barrier, but how they do so remains an area of speculation. We propose that proteases, such as the trypanopains and oligopeptidases that are released by trypanosomes, could mediate in this process. The trypanosomes also possess cell-surface-associated acid phosphatases that could play a role in invasion similar to that in advancing cancer cells. Such enzymes, perhaps acting in concert, have the potential to cause tissue degradation and ease the passage of the trypanosomes through various tissues in the host, including the blood-brain barrier.


Naturally occurring mutations of the extracellular Ca2+-sensing receptor: implications for its structure and function.
The extracellular Ca2+-sensing receptor is a member of the G-protein-coupled receptor family 3 in which agonists bind to a dimeric Venus-flytrap domain in the extracellular portion of the receptor. How agonist binding to this domain leads to activation of the seven-transmembrane domain is a major unresolved question. Information derived from the three-dimensional structure of the Venus-flytrap domain of the related metabotropic glutamate type 1 receptor, and from naturally occurring mutations of the Ca2+-sensing receptor identified in subjects with familial hypocalciuric hypercalcemia and autosomal dominant hypocalcemia offers new insights into the mechanism of receptor activation, and into the mechanism of action of allosteric modulators of the receptor.


Targeted therapy using nanocomposite delivery systems in cancer treatment: highlighting miR34a regulation for clinical applications.
The clinical application of microRNAs in modern therapeutics holds great promise to uncover molecular limitations and conquer the unbeatable castle of cancer metastasis. miRNAs play a decisive role that regulating gene expression at the post-transcription level while controlling both the stability and translation capacity of mRNAs. Specifically, miR34a is a master regulator of the tumor suppressor gene, cancer progression, stemness, and drug resistance at the cell level in p53-dependent and independent signaling. With changing, trends in nanotechnology, in particular with the revolution in the field of nanomedicine, nano drug delivery systems have emerged as a prominent strategy in clinical practices coupled with miR34a delivery. Recently, it has been observed that forced miR34a expression in human cancer cell lines and model organisms limits cell proliferation and metastasis by targeting several signaling cascades, with various studies endorsing that miR34a deregulation in cancer cells modulates apoptosis and thus requires targeted nano-delivery systems for cancer treatment. In this sense, the present review aims to provide an overview of the clinical applications of miR34a regulation in targeted therapy of cancer.


Lead contamination in Eugenia dyeriana herbal preparations from different commercial sources in Malaysia.
The Drug Control Authority (DCA) of Malaysia implemented the phase three registration of traditional medicines on 1 January, 1992. A total of 100 products in various pharmaceutical dosage forms of a herbal preparation, containing Eugenia dyeriana, either single or combined preparations (more than one medicinal plant), were analyzed for the presence of lead contamination, using atomic absorption spectrophotometry. These samples were bought from different commercial sources in the Malaysian market, after performing a simple random sampling. Results showed that 22% of the above products failed to comply with the quality requirement for traditional medicines in Malaysia. Although this study showed that 78% of the products fully complied with the quality requirement for traditional medicines in Malaysia pertaining to lead, however, they cannot be assumed safe from lead contamination because of batch-to-batch inconsistency.


Method for indirectly estimating gene-environment effect modification and power given only genotype frequency and odds ratio of environmental exposure.
Both genes and environment are important determinants of disease. In this paper we model gene-environment effect modification on the odds ratio scale OR(GEID) and show how to indirectly estimate the effect and 95% confidence intervals (CI) for the simple case of no main genetic and environmental effects [i.e., OR(GE/D) = OR(GE/D) = 1]. A statistic is presented to test the null hypothesis OR(GE/D) = 1 and to calculate corresponding power, given the odds ratio for environmental exposure OR(E/D) and population genotype frequency (g). Direct extension of the above model provides a mathematical framework for estimating confidence bounds in more complex cases involving partial genetic and/or environmental effects.


Engineering of genetic control tools in Synechocystis sp. PCC 6803 using rational design techniques.
Cyanobacteria show promise as photosynthetic microbial factories capable of harnessing sunlight and CO2 to produce valuable end products, but few genetic control tools have been characterized and utilized in these organisms. To develop a suite of control elements capable of gene control at a variety of expression strengths, a library of 10 promoter-constructs were developed and built via rational design techniques by adding individual nucleotides in a step-wise manner within the -10 and -35 cis-acting regions of the tac promoter. This suite produced a dynamic range of expression strength, exhibiting a 78 fold change between the lowest expressing promoter, Psca8- and the highest expressing promoter, Psca3-2 when tested within Synechocystis sp. PCC 6803. Additionally, this study details the construction of a chemically inducible construct for use in Synechocystis that is based on the tac repressor system most commonly used in Escherichia coli. This research demonstrates the construction of a highly expressed inducible promoter that is also capable of high levels of gene repression. Upon chemical induction with IPTG, this same mutant strain was capable of exhibiting an average 24X increase in GFP expression over that of the repressed state.


Metabolomics and molecular docking-directed antiarthritic study of the ethyl acetate extract from Celastrus orbiculatus Thunb.
Celastrus orbiculatus Thunb., an important folk medicine, has long been used for the treatment of rheumatoid arthritis and its ethyl acetate extract (COE) has been reported to possess anticancer, antiinflammation and antiarthritic effects. However, the therapeutic effect and mechanism of COE treatment in rheumatoid arthritis has been rarely studied especially from the perspective of metabolomics. To reveal the therapeutic effects of COE on adjuvant-induced arthritis (AIA) rats through histopathological analysis, non-targeted metabolomics, and molecular docking study. Forty-three Wistar rats were randomly divided into normal group, AIA model group, methotrexate group, and COE groups (80 mg/kg, 160 mg/kg and 320 mg/kg of ethyl acetate extract). Paw swelling and arthritis score were monitored through the experiment. Serum levels of tumor necrosis factor α (TNF-α) and nitric oxide were determined and histopathological evaluation was performed. Furthermore, Ultra-high performance liquid chromatography-linear trap quadrupole-Orbitrap-based metabolomics was employed to characterize metabolic changes of AIA rats after COE treatment and molecular docking was performed to predict the potential phytochemicals of COE against TNF-α. COE at three dosages could significantly relieve paw swelling and reduce arthritis scores of AIA rat. Histopathological analysis revealed remarkable decrease in synovial inflammation and bone erosion after COE treatment, especially at middle and high dosage. Additionally, COE down-regulated serum levels of TNF-α and nitric oxide. Serum metabolomics showed that 22 potential biomarkers for the COE treatment of AIA rats were identified, which were closely related to fatty acid metabolism, glycerophospholipid catabolism, and tryptophan metabolism. The molecular docking models predicted that olean-type triterpenes in COE may contribute most to therapeutic effects of rheumatoid arthritis through targeting TNF-α. COE could significantly relieve the arthritic symptoms in AIA rats and the ultra-high performance liquid chromatography-mass spectrometry based metabolomics proved to be an efficient method to characterize subtle metabolic changes of AIA rats after COE treatment.


Renoprotective effects of oleanolic acid and its possible mechanisms in rats with diabetic kidney disease.
Dyslipidemia and inflammation have great roles in the development of diabetic nephropathy (DN). Oleanolic acid (OA) is a natural triterpenoid that possesses multiple pharmacological properties including anti-oxidation, anti-inflammatory and hypoglycemia. In the present study, the effects of OA on diabetic kidney disease (DKD) and its underlying mechanisms were investigated in DKD rats. Twenty-five of a total thirty-five male Sprague-Dawley (SD) rats were used to establish for Type 2 diabetes mellitus (T2DM) model by high-fat diet combined with streptozotocin (STZ). Then rats were randomly assigned into four group: control group (n = 10), T2DM group (n = 9), OA (50 mg/kg) group (n = 7), OA (100 mg/kg) group (n = 8). Rats were sacrificed at the end of 18 weeks after feeding by intraperitoneal injection of pentobarbital sodium. Body weight (BW), fasting blood glucose (FBG), kidney weight (KW), serum lipid, 24-h urinary microalbumin (UMA), serum creatinine (Scr) and uric acid (UA) were measured. Histopathological changes were observed by PAS staining and electron microscope. The expressions of nephrin, CD68, Collagen-IV, AMPK, p-AMPK, PGC-1α, TLR4, NF-κB and TGF-β1 in kidney were also detected by immunohistochemistry or western blot. OA significantly decreased the levels of FBG, kidney index (KI), serum lipid levels, 24 h UMA, Scr, UA in diabetic rats. Additionally, OA obviously attenuated renal lipid accumulation and renal structure abnormalities in diabetic rats. Furthermore, the expression levels of nephrin, p-AMPK/AMPK, PGC-1α were elevated, while CD68, Collagen-IV, TLR4, NF-κB and TGF-β1 expressions were decreased in renal tissues of OA treated diabetic rats. OA showed dose-independent. OA can alleviate renal injury in diabetic rats through improving lipid metabolism and inflammation via AMPK/PGC-1α and TLR4/NF-κB signaling pathway.


Production of medium-sized chitosan oligomers using molecular sieves and their antibacterial activity.
Chitosan, derived from the natural polysaccharide chitin, was fragmented in very dilute acetic acid solutions using zeolites and molecular sieves, a type of zeolites, under variable reaction conditions of temperature, acid concentration, duration of reaction, and zeolites of variable pore sizes. Fragmentation resulted in the formation of appreciable amounts of chitosan oligomers comprised of 4-8 units, which were studied by using LC-MS, MS, as well as IR spectroscopy. The prepared fragments were tested for their biological activity and some of them showed antibacterial activity against Gram-positive bacteria.


Rhodium-Catalyzed Site-Selective ortho-C-H Activation: Enone Carbonyl Directed Hydroarylation of Maleimides.
Enone carbonyl directed 1,4-addition of ortho-C-H bond in chalcones to maleimides was developed under the catalysis of Rh(III). This reaction furnished a variety of chalcone-based pharmacologically useful 3-arylated succinimide derivatives in good yields with excellent selectivity.


Mechanism of cardioprotective effect of adenocine and non-glycoside cardiotonic drugs during experimental chronic cardiac insufficiency.
The therapeutic action of adenocine during cardiac insufficiency (heart failure) caused by ischemic (stenosis) or reperfusion (removal of ligature) injury to the myocardium prevents depletion of ATP, the major energy source for myocytes in the right and left ventricles, and a drop in NAD/NADH ratio. The development of energy shortage during heart failure cannot be eliminated by β-acetyldigoxin, levosimendan, or milrinone: the content of ATP in the right and left ventricular myocardium remained below the normal level by 28 and 29%, 37 and 33%, 32 and 28%, respectively; the NAD/NADH ratio of the energy supply system in cardiomyocytes did not return to normal. Adenocine increased the content of NAD to the normal level in both the right and left ventricles, while it remained below the normal level after administration of β-acetyldigoxin (by 24 and 19.5%, respectively), levosimendan (by 27 and 29%), and milrinone (by 26 and 24%). In contrast to β-acetyldigoxin, levosimendan, and milrinone, adenocine inhibited activity of poly(ADP-ribose) polymerase in both ventricles. It is concluded that adenocine directly inhibits the key enzyme triggering apoptosis; we also hypothesized that this drug activates the regulatory and signal mechanisms arresting apoptotic alterations in the myocardium during heart failure.


Second Primary Neoplasms in Patients With Uveal Melanoma: A SEER Database Analysis.
To determine the risk of second primary neoplasms (SPNs) in subjects previously diagnosed with uveal melanoma (UM), including an analysis on whether radiotherapy is a risk factor to develop these SPNs. Retrospective cohort study. Using the Surveillance, Epidemiology, and End Results (SEER) 9 database, we identified patients diagnosed with UM as their first malignancy between 1973 and 2011 (n = 3976). We obtained standardized incidence ratios (SIR) and excess absolute risks of SPNs on patients with UM compared to a reference population. Multivariate Cox regression models were used to evaluate the effect of radiotherapy in SPN risk. Sixteen percent (n = 641) of the patients developed SPNs during a median follow-up of 83 months (range, 1-463 months). This represented an 11% excess risk compared to the reference population, mainly owing to a significantly increased risk of skin melanomas (SIR = 2.93, 95% CI: 2.23-3.78) and kidney tumors (SIR = 1.91, 95% CI: 1.27-2.76), primarily in those diagnosed between 30 and 59 years of age. The occurrence of second UM was also increased (SIR = 16.90, 95% CI: 9.00-28.90), which likely includes recurrences misclassified as a second cancer. Radiotherapy was performed in 39% (n = 1538) of the patients. Multivariate analysis revealed that this treatment was not an independent risk factor for SPNs (hazard ratio = 1.06, 95% CI: 0.88-1.26, P = .54). Patients with UM presented an 11% higher risk of SPNs compared to the reference population. Radiotherapy does not seem to be a risk factor. SPNs should be considered in the surveillance of UM.


Triggering receptor expressed on myeloid cells-1 (TREM-1) inhibition in atherosclerosis.
Triggering receptor expressed on myeloid cells-1 (TREM-1) is a transmembrane protein expressed on endothelial cells, white blood cells, smooth muscle cells and platelets. TREM-1 plays an important role in innate immunity. TREM-1 activation pathways are implicated both in sepsis and in non-infectious inflammatory conditions, including atherosclerosis. TREM-1 enhances the subendothelial lipid accumulation and expression of pro-inflammatory cytokines and matrix-degrading enzymes, thereby promoting inflammation and plaque destabilization. TREM-1 inhibitors attenuate the inflammatory process in the atherosclerotic plaque, leading to plaque stabilization. This review focuses on the role of TREM-1 in the pathophysiology of atherosclerosis and the effects of TREM-1 inhibition in the natural history of the disease.


A nomogram based on clinical factors to predict calendar year readmission in patients with ulcerative colitis.
Readmission shortly after discharge is indicative of an increased disease severity for patients with ulcerative colitis (UC) and ineffectiveness to medical therapy, which may contribute to a dismal prognosis. This study aimed to explore prognostic variables with a nomogram to predict unplanned UC-related readmission within 1 year after discharge. A retrospective cohort study. Electronic medical records of all UC patients treated at our center between 1 January 2014 and 31 June 2021 were reviewed. A comprehensive analysis of various characteristics, such as demographics, comorbidities, medical history, follow-up appointments, admission endoscopy, histopathologic features, etc., was used to determine the primary end point, which was unplanned UC-related calendar year readmission. We found that the unplanned UC-related readmission rate within 1 year was 20.8%. In multivariable cox analysis, the predictors of the Elixhauser comorbidity index [Hazard ratio (HR): 3.50, 95% confidence interval (CI): 1.93-6.37], regular follow-up (HR: 0.29, 95% CI: 0.16-0.53), any history of corticosteroid use (HR: 3.38, 95% CI: 1.83-6.27), seral level of C-reactive protein (HR: 1.01, 95% CI: 1.00-1.02), and the UC endoscopic index of severity (HR: 1.29, 95% CI: 1.05-1.57) independently predicted calendar year readmission after discharge. The established nomogram had a consistently high accuracy in predicting calendar year readmission in the training cohort, with a concordance index of 0.784, 0.825, and 0.837 at 13, 26, and 52 weeks, respectively, which was validated in both the internal and external validation cohorts. Therefore, UC patients were divided into clinically low-, high-, and extremely high-risk groups for readmission, based on the calculated score of 272.5 and 378. The established nomogram showed good discrimination and calibration powers in predicting calendar year readmission in high-risk UC patients, who may need intensive treatment and regular outpatient visits.


Eosinophilic fasciitis during pregnancy.
The first reported case of eosinophilic fasciitis developing in pregnancy is discussed, and the obstetric management is reported. Eosinophilic fasciitis is a rare disease characterized by pain, swelling, and tenderness over the extremities, followed by induration of the skin. Laboratory findings include peripheral eosinophilia, hypergammaglobulinemia, an elevated erythrocyte sedimentation rate, distinctive histopathologic changes, and scleroderma-like skin induration without rheumatoid serologic markers. Diagnosis is made by biopsy of the deep fascia of the affected area. The deep fascia will be infiltrated with plasma cells, lymphocytes, and eosinophils. Prednisone is the therapy of choice. Carpal tunnel syndrome and rare serious hematologic abnormalities have been associated with eosinophilic fasciitis.


P50 suppression in human discrimination fear conditioning paradigm using danger and safety signals.
Auditory P50 suppression, which is assessed using a paired auditory stimuli (S1 and S2) paradigm to record the P50 mid-latency evoked potential, is assumed to reflect sensory gating. Recently, P50 suppression deficits were observed in patients with anxiety disorders, including panic disorder, post-traumatic stress disorder and obsessive-compulsive disorder, as we previously reported. The processes of fear conditioning are thought to play a role in the pathophysiology of anxiety disorders. In addition, we found that the P50 sensory gating mechanism might be physiologically associated with fear conditioning and extinction in a simple human fear-conditioning paradigm that involved a light signal as a conditioned stimulus (CS+). Our objective was to investigate the different patterns of P50 suppression in a discrimination fear-conditioning paradigm with both a CS+ (danger signal) and a CS- (safety signal). Twenty healthy volunteers were recruited. We measured the auditory P50 suppression in the control (baseline) phase, in the fear-acquisition phase, and in the fear-extinction phase using a discrimination fear-conditioning paradigm. Two-way (CSs vs. phase) Analysis of variance with repeated measures demonstrated a significant interaction between the two factors. Post-hoc LSD analysis indicated that the P50 S2/S1 ratio in the CS+ acquisition phase was significantly higher than that in the CS- acquisition phase. These results suggest that the auditory P50 sensory gating might differ according to the cognition of the properties (potentially dangerous or safe) of the perceived signal.


Use of a novel dual-action clip for closure of complex endoscopic resection defects.
Video 1Use of dual-action clips and clips with added atraumatic teeth to close complex endoscopic resection defects.


Unilateral biportal endoscopic technique combined with percutaneous transpedicular screw fixation for thoracolumbar burst fractures with neurological symptoms: technical note and preliminary report.
Previous studies on thoracolumbar fractures with neurological symptoms have focused on how to achieve satisfactory fracture reduction, adequate nerve decompression, and stable spinal alignment. With the development of the minimally invasive spine surgery technique, achieving satisfactory treatment results and reducing iatrogenic trauma at the same time has become a new goal of spinal surgery. This research used percutaneous transpedicular screw distraction to partially reduce the fractured vertebrae, followed by completing nerve decompression and reducing residual displacement bone fragments with the assistance of the unilateral biportal endoscopic (UBE) technique to achieve full protection of bone-ligament tissue and obtain good clinical efficacy. Guide wires were safely inserted into the fractured vertebra and adjacent upper and lower vertebra under the surveillance of anteroposterior and lateral X-ray fluoroscopy. Transpedicular screws were implanted via guide wires on the side with mild neurological deficits or bone fragment compression (the opposite side of the endoscopic operation). A titanium rod was installed and moderately distracted to reduce the fractured vertebra. Then, under the guidance of the endoscopic view, the laminectomy and ligamentum flavum resection were completed according to the position of the protruding bone fragment into the spinal canal, and the compressed dural sac or nerve root was fully exposed and decompressed. An L-shaped replacer was used to reduce residual bone fragments. The ipsilateral transpedicular screws and rod were installed and adjusted to match the contralateral side. The drainage tube was indwelled, and the incision was closed. The preoperative and postoperative images of the patients were evaluated, and the recovery of neurological symptoms was observed. Surgery was successfully completed on all six patients, and no intraoperative conversion to open surgery was performed. Postoperative images showed good reduction of the protruding bone fragment and good placement of all screws. At the last follow-up, the neurological symptoms of all patients returned to normal. The UBE technique combined with percutaneous transpedicular screw fixation in the treatment of thoracolumbar fractures with neurological symptoms can effectively achieve the reduction of displaced bone fragments, improve damaged nerve function, stabilize spinal alignment, and protect the integrity of bone-ligament tissue.


The results of radiotherapy for ependymomas: the Mayo Clinic experience.
This analysis was performed to examine the outcome of patients with histologically confirmed ependymomas of the brain or spinal cord who received postoperative radiotherapy. Eighty patients with histologically confirmed ependymomas were evaluated retrospectively. All were treated with various combinations of surgery, radiotherapy (RT), and chemotherapy. Follow-up ranged from 5 to 30 years (median 10.4 years). The 5- and 10-year survival rates for the entire study group were 79% and 73%, respectively. Patients with low-grade (1 and 2 of 4) tumors had a 5-year survival rate of 87% as compared to 27% for those with high-grade (3 and 4 of 4) tumors (p < 0.0001). Patients with tumors of the spine had a 5-year survival rate of 97% as compared to 68% for those with infratentorial tumors, and 62% for those with supratentorial tumors (p = 0.03). Patients with myxopapillary ependymomas of the spine had a 5-year survival rate of 100% as compared with 76% for patients with other histological subtypes of ependymoma (p = 0.02). Multivariate analysis revealed that the survival rate was independently associated with tumor grade (p = 0.0007) and histological subtype (p = 0.02). Twenty-eight patients (35%) experienced local failure and 10 patients (13%) developed leptomeningeal seeding. The 5-year leptomeningeal failure rate was 10% in patients with low-grade tumors as compared to 41% for patients with high grade tumors (p = 0.01). Patients with low-grade tumors, especially those with myxopapillary subtypes, have high 5-year survival rates when treated with post-operative radiotherapy. High grade ependymomas are associated with a much poorer outcome. New forms of therapy are required to improve the outcome of patients with high-grade ependymomas.


Characteristics and mechanism of glutamine-dipeptide absorption in human intestine.
Using in vivo and in vitro techniques, the mechanism by which intestinal mucosa obtains glutamine from luminal oligopeptides was investigated in humans. The rate of hydrolysis by mucosal brush border membrane was more than threefold greater for alanylglutamine than for glycylglutamine. Despite this difference, rates of dipeptide and amino acid disappearance during intestinal perfusion were greater from test solutions containing glycylglutamine than alanylglutamine. Furthermore, rates of intraluminal appearance of products of hydrolysis during the infusion of two dipeptides were similar and less than 5% of the disappearance rate of the parent dipeptide. In contrast to free glutamine, uptake of peptide-bound glutamine by brush border membrane vesicles was not inhibited by deletion of sodium or addition of free amino acids to the incubation medium but was inhibited by other oligopeptides and stimulated by a proton gradient. Inhibition constants for the saturable uptake of glycylglutamine and alanylglutamine by vesicles were not significantly different, suggesting similar affinities for the peptide transporter. It is concluded that in human intestine the predominant mechanism for assimilation of glutamine-dipeptides is absorption as intact dipeptide rather than hydrolysis.


Rate and age-dependent damage elasticity formulation for efficient hip fracture simulations.
Prediction of bone failure is beneficial in a range of clinical situations from screening of osteoporotic patients with high fracture risk to assessment of protective equipment against trauma. Computational efficiency is an important feature to consider when developing failure models for iterative applications, such as patient-specific diagnosis or design of orthopaedic devices. The authors previously developed a methodology to generate efficient mesoscale structural full bone models. The aim of this study was to implement a damage elasticity formulation representative of an elasto-plastic material model with age and strain rate dependencies compatible with these structural models. This material model was assessed in the prediction of femoral fractures in longitudinal compression and side fall scenarios. The simulations predicted failure loads and fracture patterns in good agreement with reported results from experimental studies. The observed influence of strain rate on failure load was consistent with literature. The superiority of a simplified elasto-plastic formulation over an elasto-brittle bone material model was assessed. This computationally efficient damage elasticity formulation was capable of capturing fracture development after onset.


Use of US Food and Drug Administration Expedited Drug Development and Review Programs by Orphan and Nonorphan Novel Drugs Approved From 2008 to 2021.
The US Food and Drug Administration (FDA) has 4 programs that can be used alone or in combination to expedite drug availability: Accelerated Approval, Breakthrough Therapy, Fast Track, and Priority Review. Drugs using these programs can include novel drugs, which do not contain a previously FDA-approved active moiety, and orphan drugs, intended for diseases or conditions affecting fewer than 200 000 people; to date, no comprehensive evaluation of how these programs have been used in combination has been published. To assess how often and in what combinations expedited programs are used in the development and review of approved novel biologics and small-molecule drugs, stratified by orphan drug status and indication. This cross-sectional study evaluated all novel drugs that were FDA approved between January 1, 2008, and December 31, 2021. The main outcome was the frequency with which expedited programs were used and in what combinations, stratified by orphan drug status and drug type (small molecule vs therapeutic biologic). The unit of analysis was the novel drug-indication pair because a drug can be approved for multiple indications, each of which may use a different expedited program or differ in orphan drug status. The study included 581 novel drug-indication pairs approved during the 14-year study period; 252 (43.4%) were orphan drugs, 139 (23.9%) were therapeutic biologics, and 442 (76.1%) were small-molecule drugs. Use of at least 1 expedited program increased from 11 of 26 drug-indication pairs (42.3%) in 2008 to 41 of 55 (74.5%) in 2021. Of the 363 approved drug-indication pairs using at least 1 expedited program, 225 (62.0%) were orphan drugs; at least 1 expedited program was used by 97 of the 139 approved biologic drugs (69.8%) and by 266 of the 442 approved small-molecule drugs (60.2%). Eighty-two of the 581 novel drug-indication pairs (14.1%) used the Accelerated Approval Program; of those, 65 (79.3%) were oncology drugs and 70 (85.4%) had an orphan designation. The study showed that use of the FDA's expedited programs to bring novel drugs to market in the US increased from 2008 to 2021. The findings suggest that this trend is likely to continue.


Estimating testosterone concentrations in adolescent girls: Comparison of two direct immunoassays to liquid chromatography-tandem mass spectrometry.
The appropriate role of direct total testosterone (T) immunoassays in reproductive research is controversial. To assess the concordance between two direct immunoassays and a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for total T in adolescent girls with measured concentrations < 50 ng/dl. Cross-sectional analysis. Academic medical center. Adolescent girls (age 8.4-18.1 years) participating in clinical research protocols. Paired blood samples were obtained for total T by LC-MS/MS (n = 66; Mayo Clinic Laboratory) and by direct immunoassay (Center for Research in Reproduction)-either radioimmunoassay (RIA; n = 31) or chemiluminescence immunoassay (CLIA; n = 35). At the time of assay, laboratories were unaware that results would be compared. Measurement agreement between immunoassay and LC-MS/MS. Measured T concentrations (LC-MS/MS) were <7 to 44 ng/dl. The average difference between RIA and LC-MS/MS was 0.84 [-0.89, 2.56] ng/dl (mean [95% confidence interval]). RIA correlated very strongly with LC-MS/MS (r = 0.899; p < 0.0001); and both Deming regression and Bland-Altman analysis suggested no bias. The average difference between chemiluminescence and LC-MS/MS was 1.39 [-0.83, 3.60] ng/dl. CLIA correlated strongly with LC-MS/MS (r = 0.806; p < 0.0001). While Bland-Altman analysis suggested no systematic bias, Deming regression analysis suggested that, as measured values increased, values obtained by CLIA tended to be progressively, albeit only modestly, higher than those obtained by LC-MS/MS. These data support the use of rigorously-performed and carefully-validated direct T immunoassays in high-quality endocrine research in peripubertal adolescent girls.


Decade of aortic valve sparing reimplantation: are we pushing the limits too far?
This single center study assesses the outcome of aortic valve sparing reimplantation (AVS) in 284 consecutive patients who were operated on for various indications during the last 11 years. From July, 1993, to July, 2004, 284 patients underwent AVS. Mean age was 53+/-16 (range 8 to 84) years. Of the 284 patients, 184 were male (64.8%) and 54 (19%) experienced Marfan's syndrome. Acute aortic dissection Stanford type A was present in 53 patients (19%) and a bicuspid aortic valve was present in 17 patients (6%). Concomitant arch replacement was necessary in 120 patients (42%). Additional procedures were performed in 66 patients (23.2%). Mean follow-up time was 41+/-32 (range 0 to 130) months. The 30-day mortality was 3.2% overall, 11.3% in emergency patients, and 1.3% in elective patients. Mean bypass time was 174+/-48 (range 90 to 440) minutes and aortic cross clamp time was 132+/-33 (range 64 to 283) minutes. In patients undergoing arch replacement, circulatory arrest was 25+/-17 (range 7 to 99) minutes. Rethoracotomy for bleeding was required in 4.6% of patients. During follow-up, there were 20 (7.3%) late deaths. Reoperation of the reconstructed valve was required in 15 patients (5.3%); underlying reasons were endocarditis (n=4) and aortic insufficiency (n=11) requiring aortic valve replacement. Average grade of aortic insufficiency increased significantly from 0.23+/-0.46 postoperatively to 0.34+/-0.59 at latest evaluation (P=0.026). Two patients experienced a transient ischemic attack early postoperatively; no further thromboembolic complications were noticed. The majority of patients (96%) presented with a favorable exercise tolerance at last contact. The aortic valve reimplantation technique leads to excellent clinical outcome in patients with various pathologies. Lack of anticoagulation and favorable durability should encourage the extension of indications for this technique.


The spatial frequency and orientation selectivity of the mechanisms that extract motion-defined contours.
The human visual system can undertake a specialized form of motion integration, one that enables the presence of extended spatial contours to be disambiguated from their backgrounds. We have shown previously that the visual system can selectively integrate local motion signals when their directions are along spatial contours and its efficiency is inversely related to the curvature of the contour involved (Ledgeway, T., & Hess, R. F. (2002). Vision Research, 42, 653-659). This integration primarily involves the direction, rather than the speed, of local motion signals. In the present study, we sought to investigate both the spatial frequency and orientation tuning of this specialized contour integration process, using a path detection paradigm. The results show that the tuning for spatial frequency is very broad, in line with previous studies that have examined this issue. In contrast, the orientation selectivity of the mechanism mediating contour extraction under these conditions is relatively narrowband. Thus, spatial frequency but not orientation pooling appears to take place prior to the extraction of motion-defined contours, a situation that is different from that previously shown for spatial contours composed of static, oriented elements.


New amphibian models for the study of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP).
We report the development of two animal models in amphibians (frogs and salamanders) in whom 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) produces the behavioral (neurological) and biochemical equivalents of the human disease and, in addition, a measurable modification in at least one form of pigment-bearing cell from the neural crest, the skin melanocyte. We propose that this new approach can become an inexpensive, easily quantifiable model for the study of the effect of MPTP on the central and peripheral nervous systems. We also demonstrate that the toxic effect of MPTP can be completely abolished in vivo by treatment with a monoamine oxidase inhibitor and potentiated by an inhibitor of catechol-O-methyltransferase. MPTP is catabolised by oxidation into toxic metabolites, but 1-methyl-4-phenylpyridinium ion (MPP+), the proposed end-metabolite, is even more toxic than MPTP in this model, possibly through a different mechanism.


Antigenic determinants shared by lysosomal proteins of Dictyostelium discoideum. Characterization using monoclonal antibodies and isolation of mutations affecting the determinant.
Polyclonal antisera raised against a single lysosomal enzyme from Dictyostelium discoideum will cross-react with other lysosomal proteins due to the presence of common post-translational modification antigens. We have now isolated hybridoma cell lines secreting monoclonal antibodies that also recognize a shared lysosomal determinant. Although the same proteins are recognized by the monoclonal and polyclonal antibodies, the actual determinants differ. Also, the polyclonal serum contains antibodies which recognize at least one determinant not recognized by the monoclonal antibodies. All of the antigenic proteins are minor cell proteins but represent a majority of the proteins secreted from the cells during axenic growth. Anode-directed nonequilibrium two-dimensional gel electrophoresis of cellular and secreted protein indicates that the antigenic proteins have extremely acidic pI values; moreover, most are both sulfated and phosphorylated. Several sulfated polysaccharides, including dextran sulfate, chondroitin sulfate, and heparin inhibit binding of the monoclonal but not the polyclonal antibodies, indicating that the shared determinant recognized by the monoclonal antibodies may involve sulfated oligosaccharides. As a step in analyzing the function of the post-translational modification of lysosomal enzymes, we have screened a mutagenized population of amoeba for those lacking the antigenic determinant recognized by one of the monoclonal antibodies. Five cell lines have been isolated that completely lack the determinant. They appear to fall into at least three different classes based upon physical properties of the enzymes. One strain lacks any detectable sulfation of glycoproteins. Several of the strains are capable of forming fruiting bodies indicating that the antigenic determinant plays no crucial role in morphogenesis under laboratory conditions.


Tetrandrine induces lipid accumulation through blockade of autophagy in a hepatic stellate cell line.
Macroautophagy, or autophagy, is a cellular response in which unnecessary cytoplasmic components, including lipids and organelles, are self-degraded. Recent studies closely related autophagy to activation of hepatic stellate cells (HSCs), a process critical in the pathogenesis of liver fibrosis. During HSC activation, cytoplasmic lipid droplets (LDs) are degraded as autophagic cargo, and then cells express fibrogenic genes. Thus, inhibition of autophagy in HSCs is a potential therapeutic approach for attenuating liver fibrosis. We found that tetrandrine, a bisbenzylisoquinoline alkaloid isolated from Stephania tetrandra, induced lipid accumulation, a phenotype associated with quiescent HSCs, through blockade of autophagy in the rat-derived HSC line HSC-T6. Tetrandrine inhibited autophagic flux without affecting lysosomal function. A phenotypic comparison using siRNA knockdown suggested that tetrandrine may target regulators, involved in fusion between autophagosomes and lysosomes (e.g., syntaxin 17). Moreover, perilipin 1, an LD-coated protein, co-localized specifically with LC3, a marker protein for autophagosomes, in tetrandrine-treated HSC-T6 cells. This suggests a potential role for perilipin 1 in autophagy-mediated LD degradation in HSCs. Our results identified tetrandrine as a potential tool for prevention and treatment of HSC activation.


Panendoscopy and synchronous second primary tumors in head and neck cancer patients.
Endoscopy techniques are used to diagnose and to determine the extent and exact location of malignancies in the head and neck region, bronchial tree and esophagus. Panendoscopy is used to find the primary tumor in the case of metastatic disease from unknown primaries or to detect a simultaneous second primary tumor at the time of diagnosis of a malignancy in the upper aerodigestive tract (UADT). The value of panendoscopy has been debated lately because of the relatively small proportion of malignant findings and because of the lack of convincing data concerning its effect on survival rates. However, despite the relatively low proportion of positive findings, their significance is often crucial for the individual patient. The significant number of late metachronous, second primaries, especially in the lungs, also emphasizes the importance of follow-up endoscopies. This study consists of 203 consecutive patients with squamous cell cancer (SCC) of the upper aerodigestive tract who underwent panendoscopy in Turku University Central Hospital as part of the initial diagnostic workup from 1992-1999. Eight patients with synchronous second primaries were found to represent a prevalence of 3.9%, and in addition, 19 patients with metachronous tumors were diagnosed. In the case reports we illustrate the importance of some of these findings.


Students' perceptions of school climate during the middle school years: associations with trajectories of psychological and behavioral adjustment.
A cross-domain latent growth curve model was used to examine the trajectories of change in student perceptions of four critical dimensions of school climate (i.e., teacher support, peer support, student autonomy in the classroom, and clarity and consistency in school rules and regulations) among 1,451 early adolescents from the beginning of sixth through the end of eighth grade; and the effects of such trajectories on the rate of change in psychological and behavioral adjustment. Findings indicated that all of the dimensions of perceived school climate declined over the 3 years of middle school. Furthermore, declines in each of the dimensions of perceived school climate were associated with declines over time in psychological and behavioral adjustment. Moreover, the direction of effects between each dimension of perceived school climate and psychological or behavioral adjustment were often unidirectional rather than bi-directional, underscoring the role of perceived school climate in the psychological and behavioral health of early adolescents. Gender and socioeconomic class differences in these patterns are noted.


Total hip replacement ad modum Lubinus: five- to seven-year follow-up.
A study of 141 total hip replacements with the Lubinus prosthesis is presented. The complications in the total patient group and the results in 129 hips with follow-up times from 5 to 7 years are described. Three prostheses (2.1%) have been reoperated because of loosening. There were radiographic signs of definitive loosening in 2.3%. Varus positioning was associated with an increased loosening rate. No infections have been encountered, and none of the hips have undergone excision arthroplasty. Ninety-two percent of the hips were free from significant pain and 78% had a total range of motion exceeding 160 degrees. THR was performed, without subsequent infections, in a conventional operating room using prophylactic penicillin and gentamicin cement. We propose that the surgical technique of exposure and the cement injection technique with a partial vacuum in the medullary canal may be responsible for the low incidence of femoral prosthesis loosening (0.7%).


Anaerobic bloodstream infections in Italy (ITANAEROBY): A 5-year retrospective nationwide survey.
A lack of updated data on the burden and profile of anaerobic bloodstream infections (ABIs) exists. We assessed the incidence of ABIs and trends in antimicrobial resistance in anaerobes isolated from blood in Italy. We conducted a retrospective study on 17 Italian hospitals (2016-2020). Anaerobes isolated from blood culture and their in vitro susceptibility profiles (EUCAST-interpreted) were registered and analyzed. A total of 1960 ABIs were identified. The mean age of ABIs patients was 68.6 ± 18.5 years, 57.6% were males. The overall incidence rate of ABIs was 1.01 per 10.000 patient-days. Forty-seven% of ABIs occurred in medical wards, 17% in ICUs, 14% in surgical wards, 7% in hemato-oncology, 14% in outpatients. The three most common anti-anaerobic tested drugs were metronidazole (92%), clindamycin (89%) and amoxicillin/clavulanate (83%). The three most common isolated anaerobes were Bacteroides fragilis (n = 529), Cutibacterium acnes (n = 262) and Clostridium perfringens (n = 134). The lowest resistance rate (1.5%) was to carbapenems, whereas the highest rate (51%) was to penicillin. Clindamycin resistance was >20% for Bacteroides spp., Prevotella spp. and Clostridium spp. Metronidazole resistance was 9.2% after excluding C. acnes and Actinomyces spp. Bacteroides spp. showed an increased prevalence of clindamycin resistance through the study period: 19% in 2016, 33% in 2020 (p ≤ 0.001). Our data provide a comprehensive overview of the epidemiology of ABIs in Italy, filling a gap that has existed since 1995. Caution is needed when clindamycin is used as empirical anti-anaerobic drug.


Conventional versus 'all-inside' anterior cruciate ligament reconstruction: a randomized controlled trial comparing hamstring strength and functional outcome.
Cite this article: Bone Joint Open 2020;1-11:706-708.


[Patient-reported outcome measures for assessing health-related quality of life in patients with voice and swallowing disorders].
Standardized and validated patient surveys in the form of questionnaires are becoming increasingly important in otorhinolaryngology. Subjective symptoms and functional limitations but also health-related quality of life are assessed with so-called patient-reported outcome measures (PROMs). Besides patient history and objective findings, these questionnaires can be applied as another important source for ensuring the quality of patient-centered care. In German-speaking countries there are now several PROMs that specifically ask for the patient perspective on voice and swallowing disorders. For patients with voice disorders the following questionnaires based on the Voice Handicap Index (VHI), which are also used internationally, are mainly applied: VHI-30, VHI-12i, and VHI-9i. In the field of oropharyngeal dysphagia, the German versions of the Eating Assessment Tool-10 (EAT-10), des Sydney Swallow Questionnaire SSQ‑G, the Swallowing Quality of Life Questionnaire (SWAL-QoL), and the MD Anderson Dysphagia Inventory (MDADI) are currently mostly used.


Protective effect of serofendic acid, administered intravenously, on cerebral ischemia-reperfusion injury in rats.
Serofendic acid is a low-molecular-weight compound extracted from fetal calf serum. We previously reported that intracerebroventricular administration of serofendic acid prevents cerebral ischemia-reperfusion injury. However, the effect of peripheral administration of serofendic acid on cerebral ischemia-reperfusion injury has not been examined. In the present study, we investigated the effect of intravenous administration of serofendic acid against cerebral ischemia-reperfusion injury using transient middle cerebral artery occlusion model rats. Serofendic acid (10mg/kg) administrated three times, including 30min before the onset of ischemia, just after the onset of ischemia and just before reperfusion reduced the infarct volume and improved the neurological dysfunction induced by ischemia-reperfusion without affecting regional cerebral blood flow or physiological parameters. However, there were no protective effects when serofendic acid (30mg/kg) was only administered once at 30min before the onset of ischemia, just after the onset of ischemia, or just before reperfusion. Our results reveal the importance of maintaining the blood concentration of serofendic acid for preventing cerebral ischemia-reperfusion injury.


Regulation of ghrelin structure and membrane binding by phosphorylation.
The peptide hormone ghrelin requires Ser-3 acylation for receptor binding, orexigenic and anti-inflammatory effects. Functions of desacylghrelin are less well understood. In vitro kinase assays reveal that the evolutionarily conserved Ser-18 in the basic C-terminus is an excellent substrate for protein kinase C. Circular dichroism reveals that desacylghrelin is approximately 12% helical in aqueous solution and approximately 50% helical in trifluoroethanol. Ser-18-phosphorylation, Ser-18-Ala substitution, or Ser-3-acylation reduces the helical character in trifluoroethanol to approximately 24%. Both ghrelin and desacylghrelin bind to phosphatidylcholine:phosphatidylserine sucrose-loaded vesicles in a phosphatidylserine-dependent manner. Phosphoghrelin and phosphodesacylghrelin show greatly diminished phosphatidylserine-dependent binding. These results are consistent with binding of ghrelin and desacylghrelin to acidic lipids via the basic face of an amphipathic helix with Ser-18 phosphorylation disrupting both helical character and membrane binding.


Reversal of locked-in syndrome with anticoagulation, induced hypertension, and intravenous t-PA.
Widespread use of intravenous tissue plasminogen activator (t-PA) for acute ischemic stroke is limited by multiple contraindications to its use. This article describes a patient with stuttering symptoms of pontine ischemia caused by vertebrobasilar dissection who suddenly deteriorated into a locked-in state 32 hours after symptom onset. The quadriparesis was successfully reversed within 3 hours of onset with the combination of pharmacologically induced hypertension, anticoagulation, and intravenous t-PA. Even in the face of numerous contraindications (including hypertension, anticoagulation, and treatment beyond 3 hours of symptom onset), intravenous t-PA can be used successfully in carefully selected cases.


Summer diapause induced by high temperatures in the oriental tobacco budworm: ecological adaptation to hot summers.
Summer diapause in Helicoverpa assulta (Hübner), which prolongs the pupal stage, particularly in males, is induced by high temperatures. In the laboratory, 3(rd)-, 4(th)-, 6(th)-instar and prepupal larvae were exposed to high temperatures - 33 and 35 °C with a photoperiod of LD16:8 - until pupation to induce summer diapause. The results showed that the incidence of summer diapause was influenced by temperature, stage exposed, and sex. The higher the temperature, the more often summer diapause was attained. Sixth-instar and prepupal larvae were the sensitive stages for summer diapause induction. H. assulta summer-diapausing pupae needed diapause development to resume development when temperatures became favorable. Furthermore, both body mass and energy storage capacity (lipid and glycogen) were significantly affected by diapause rather than sex, and were significantly higher in summer-diapausing pupae than in non-diapausing pupae. In addition, the body mass loss and respiration rate showed that the rate of metabolism in the summer-diapausing pupae was consistently lower than in non-diapausing pupae, which were significantly affected by diapause and pupal age. We conclude that summer diapause in H. assulta is a true diapause, and H. assulta has evolved mechanisms to accumulate energy storage and to lower its metabolism to adapt to hot summers.


Respiratory and cuticular water loss in insects with continuous gas exchange: comparison across five ant species.
Respiratory water loss (RWL) in insects showing continuous emission of CO(2) is poorly studied because few methodologies can measure it. Comparisons of RWL between insects showing continuous and discontinuous gas exchange cycles (DGC) are therefore difficult. We used two recently developed methodologies (the hyperoxic switch and correlation between water-loss and CO(2) emission rates) to compare cuticular permeabilities and rates of RWL in five species of ants, the Argentine ant (Linepithema humile) and four common native ant competitors. Our results showed that RWL in groups of ants with moderate levels of activity and continuous gas exchange were similar across the two measurement methods, and were similar to published values on insects showing the DGC. Furthermore, ants exposed to anoxia increased their total water loss rates by 50-150%. These results suggest that spiracular control under continuous gas exchange can be as effective as the DGC in reducing RWL. Finally, the mesic-adapted Argentine ant showed significantly higher rates of water loss and cuticular permeability compared to four ant species native to dry environments. Physiological limitations may therefore be responsible for restricting the distribution of this invasive species in seasonally dry environments.


Chemically induced lipid phase separation in model membranes containing charged lipids: a spin label study.
The lipid distribution in binary mixed membranes containing charged and uncharged lipids and the effect of Ca2+ and polylysine on the lipid organization was studied by the spin label technique. Dipalmitoyl phosphatidic acid was the charged, and spin labelled dipalmitoyl lecithin was the uncharged (zwitterionic) component. The ESR spectra were analyzed in terms of the spin exchange frequency, Wex. By measuring Wex as a function of the molar percentage of labelled lecithin a distinction between a random and a heterogeneous lipid distribution could be made. It is established that mixed lecithin-phosphatidic acid membranes exhibit lipid segregation (or a miscibility gap) in the fluid state. Comparative experiments with bilayer and monolayer membranes strongly suggest a lateral lipid segregation. At low lecithin concentration, aggregates containing between 25% and 40% lecithin are formed in the fluid phosphatidic acid membrane. This phase separation in membranes containing charged lipids is understandable on the basis of the Gouy-Chapman theory of electric double layers. In dipalmitoyl lecithin and in dimyristoyl phosphatidylethanolamine membranes the labelled lecithin is randomly distributed above the phase transition and has a coefficient of lateral diffusion of D = 2.8-10(-8) cm2/s at 59 degrees C. Addition of Ca2+ dramatically increases the extent of phase separation in lecithin-phosphatidic acid membranes. This chemically (and isothermally) induced phase separation is caused by the formation of crystalline patches of the Ca2+-bound phosphatidic acid. Lecithin is squeezed out from these patches of rigid lipid. The observed dependence of Wex on the Ca2+ concentration could be interpreted quantitatively on the basis of a two-cluster model. At low lecithin and Ca2+ concentration clusters containing about 30 mol % lecithin are formed. At high lecithin or Ca2+ concentrations a second type of precipitation containing 100% lecithin starts to form in addition. A one-to-one binding of divalent ions and phosphatidic acid at pH 9 was assumed. Such a one-to-one binding at pH 9 was established for the case of Mn2+ using ESR spectroscopy. Polylysine leads to the same strong increase in the lecithin segregation as Ca2+. The transition of the phosphatidic acid bound by the polypeptide is shifted from Tt = 47.5 degrees to Tt = 62 degrees C. This finding suggests the possibility of cooperative conformational changes in the lipid matrix and in the surface proteins in biological membranes.


Squamous metaplasia in the cyst epithelium of type 1 congenital pulmonary airway malformation after thoracoamniotic shunt placement.
Thoracoamniotic shunting is the treatment of choice for management of the fetus with type 1 congenital pulmonary airway malformation. Thoracoamniotic shunting has been performed to reduce life-threatening risks such as fetal hydrops. However, caution is needed because of possible complications. Here, we report that thoracoamniotic shunting can cause histologic changes in the cyst epithelia. In 5 of 8 patients treated prenatally with thoracoamniotic shunting, squamous metaplasia in the cyst epithelia was seen; whereas squamous metaplasia was not found in 6 patients who were not treated with this procedure. Our results reveal that long-term exposure to the intrauterine environment could possibly lead to the change in the nature of cyst epithelium and consequent squamous metaplasia.


Transcriptional feedback of Neurospora circadian clock gene by phosphorylation-dependent inactivation of its transcription factor.
The circadian clock protein Frequency (FRQ) feedback-regulates its own expression by inhibiting its transcriptional activator, White Collar Complex (WCC). We present evidence that FRQ regulates the bulk of WCC through modulation of its phosphorylation status rather than via direct complex formation. In the absence of FRQ, WCC is hypophosphorylated and transcriptionally active, while WCC is hyperphosphorylated and transcriptionally inactive when FRQ is expressed. The phosphorylation status of WCC changes rhythmically over a circadian cycle. Dephosphorylation and activation of WCC depend on protein phosphatase 2A (PP2A), and WCC is a substrate of PP2A in vitro. Hypophosphorylated WCC binds to the clock box of the frq promoter even in the presence of FRQ, while binding of hyperphosphorylated WCC is compromised even when FRQ is depleted. We propose that negative feedback in the circadian clock of Neurospora is mediated by FRQ, which rhythmically promotes phosphorylation of WCC, functionally equivalent to a cyclin recruiting cyclin-dependent kinase to its targets.


Effect of methotrexate and its photodegradation products on the temperature induced denaturation of human serum albumin.
Effect of the photodegradation of chemotherapeutic agent methotrexate on the denaturation of human serum albumin has been studied by differential scanning calorimetry and fluorimetric measurements. Photoluminescence studies highlighted entropy driven binding of both methotrexate molecules and its degradation products to the albumin molecules. The calorimetric measurements evaluated by the Kissinger method show elevated activation energy in the presence of methotrexate. Similar but moderated enhancement of the activation energy was obtained in the presence of the photodegradation products of methotrexate. These results highlight stabilization of the human serum albumin by the methotrexate drug which finding may contribute to fine tuning of methotrexate applications in therapy.


The effect of intradermal administration of Corynebacterium parvum on the immune response to hepatitis Bs antigen.
The immunopotentiating effect of the intradermal administration of a course of four doses (0.25 ml) of a standard suspension of killed C. parvum (2 mg/ml) was studied in a group of 10 asymptomatic chronic hepatitis B surface antigen (HBsAg) carriers, as well as in 11 persons with antibodies to HBsAg (anti-HBs) and six without HBsAg or anti-HBs. HBsAg, anti-HBs, and leukocyte migration inhibition (LMI) studies were performed in pre- and post-inoculation blood samples. C. parvum produced a substantial increase of anti-HBs titre in persons with preexisting anti-HBs. However, anti-HBs responses were not induced in carriers. HBsAg was not eliminated and its titre remained practically unchanged in carriers. These results support the hypothesis that in carriers the specific defect in the immune response to HBsAg possibly exists at the B cell level.


Early and long-term prognostic value of Troponin-I after cardiac surgery in newborns and children.
Troponin-I (Tn-I) is a well-recognized early postoperative marker for myocardial damage in adults and children. The present prospective study was undertaken to investigate whether a postoperative Tn-I value higher than 35 microg/l is able to predict long-term outcome as it does in early postoperative course, after surgery for congenital heart defects (CHD). Five hundred and twenty patients (median age 11 months; male 54.7%: 284 patients) undergoing congenital heart defect repair on cardiopulmonary by-pass were prospectively updated in our database including postoperative Tn-I values. Seventy of them (13.4%) (mean age 2.6+/-5.8 months) (70/520) experienced low output syndrome in the early postoperative period. According to the complexity of their malformations, we have arbitrarily divided these patients into two groups: group A included atrial and ventricular septal defects (13 patients), while group B included hypoplastic left heart syndrome, atrio-ventricular canal, transposition of great vessels, tetralogy of Fallot, double outlet right ventricle, truncus arteriosus, total anomalous venous return, and other combined diseases (57 patients). These patients are the object of our study. We reviewed clinical, laboratory, and echocardiographic data performed in the immediate postoperative course (within 24 h) and in the follow-up. In this study, 13 patients died (13/70 patients; 18.5%), 12 in group B and 1 in group A. In deceased patients, mean Tn-I value was 130+/-175 microg/l (CK-MB 570+/-280 microg/l). Conversely, survivors showed a lower mean Tn-I value (25.5+/-28.9 microg/l; CK-MB 76+/-86 microg/l). Overall, Tn-I peak value was higher than 35 microg/l in 19 patients (19/70; 27.2%); among these, 9 died (median Tn-I was 163+/-186 microg/l), whereas in survivors it was 73.4+/-37 microg/l (p=0.37). The remaining four patients who died had a median Tn-I value of 21 microg/l. When Tn-I exceeded 35 microg/l (>100 microg/l in two cases), at echocardiogram a severely depressed cardiac function was evident. Nevertheless, at long-term follow-up (12+/-6 months), the echocardiogram showed an enhanced cardiac performance with an ejection fraction of 70+/-8.5% in all; none of these patients presented with worsened ventricular function. Cardiac Tn-I is a specific and sensitive marker of myocardial injury after cardiac surgery and it may predict early in-hospital outcomes. However, by long-term echocardiographic analysis, cardiac Tn-I value looses its prognostic significance and therefore it is not a predictor of long-term ventricular dysfunction.


Ventricular performance and quality of life in patients who underwent radiofrequency AV junction ablation and permanent pacemaker implantation due to medically refractory atrial tachyarrhythmias.
In patients with drug refractory atrial tachyarrhythmias and previous failed attempts of ablation of the arrhythmia substrate, radiofrequency (RF) modulation or ablation of the atrioventricular (AV) junction is an alternative procedure. Of this study was to assess the efficacy and long term results of RF AV junction ablation in conjunction with permanent pacemaker implantation, in the management of patients with drug resistant atrial tachyarrhythmias. Between 4/92 and 1/97, 46 patients (30 male, 16 female, 67 +/- 12 years) underwent RF AV junction ablation because of paroxysmal atrial fibrillation (24 patients), chronic atrial fibrillation (13 patients), atrial flutter (5 patients) and atrial tachycardia (4 patients). The underlying heart disease was dilated cardiomyopathy (16), ischemic heart disease (9), hypertensive heart disease (6), hypertrophic cardiomyopathy (3), atrial septal defect (2) and non structural heart disease (10). The duration of symptoms was 6.4 +/- 3.5 years at a maximal heart rate 169 +/- 24 bpm. The hospital admissions in the last 12 months were 8.2 +/- 3 per patient. The failed antiarrhythmic drugs were 3.5 +/- 2.1. The functional NYHA class was 2.7 +/- 0.6. Patients with atrial flutter and atrial tachycardia had previous failed attempts of RF ablation of the arrhythmia substrate. Thirty patients had a compromised left ventricular systolic function with LVEF below 50% (mean 34 +/- 9%). AV junction ablation was achieved in all patients after 4 +/- 2.5 RF applications. Post ablation, the selected pacing mode was DDD-R for the 33 patients with paroxysmal atrial tachyarrhythmias and VVI-R for the 13 pts with chronic atrial fibrillation. The dual chamber pacemakers implanted had the option of automatic mode switch. During the follow-up period of 28 +/- 13 months (6-47), AV conduction recovered in 1 patient. Antiarrhythmic treatment was necessary in only 7 patients. Post ablation the new functional NYHA class was 1.4 +/- 0.8 (p < 0.001). Post ablation hospital admissions, including ordinary pacemaker follow-up visits, were 4 +/- 1 per patient per year (p < 0.001). Six months after the procedure the LVEF of the study population was increased from 42 +/- 16% to 50 +/- 14% (p = NS). In the 30 patients with heart failure the LVEF was significantly increased to 46 +/- 8% (p < 0.05). Symptomatic relief or significant improvement was observed in all patients as showed by the answers given in a customized questionnaire before and after the procedure. In patients with drug refractory atrial tachyarrhythmias, RF AV junction ablation and permanent pacemaker implantation is an alternative therapy with excellent long term results in terms of arrhythmia control, ventricular performance and quality of life.


Inhibition of hepatic protein degradation by synthetic analogues of chymostatin.
Analogues of the microbial proteinase inhibitor chymostatin have been synthesized. The two most promising analogues were tested on protein turnover in isolated rat hepatocytes. Their effect is much similar to the effect of chymostatin, but the analogues are even more powerful inhibitors, probably due to an increased effect on lysosomal thiol proteinases. The analogues blocked most of the lysosomal (i.e. methylamine-sensitive) degradation of endogenous protein and caused a 50% inhibition of the non-lysosomal degradation; the effect occurred rapidly and was reversed upon washing the cells. One of the analogues, Z-Arg-Leu-Phe(H), is the most potent inhibitor of hepatic protein degradation so far found.


Acute IGF-1 deficiency in multiple trauma victims.
The circulating levels of IGF-1 in the catabolic flow phase of injury were measured in multiple trauma victims once before nutritional support and again after 4 days of intravenous feeding. The hypermetabolic and highly catabolic state of injury is characterized by a diminished level of IGF-1 along with increased levels of stress hormones. Inverse relationships of adiposity and aging on IGF-1 levels are seen both in injured and uninjured subjects. Feeding could restore the IGF-1 levels in non-obese, young patients but it is delayed in obese or elderly subjects. This diminished responsiveness to feeding of IGF-1 hormone deficiency in obese and elderly subjects should be taken into consideration in their post-trauma treatment and may warrant adjuvant administration in these types of patients.


Effects of lipoprotein apheresis on PCSK9 levels.
PCSK9 (Proprotein Convertase Subtilisin/Kexin Type 9) increases LDL cholesterol (LDL-C) levels by stimulating the degradation of Low Density Lipoprotein receptors (LDL-r). This protein is now of high interest because antibodies which inhibit its effect on LDL-r are being developed. A severe hypercholesterolemia and / or an elevation of lipoprotein(a) can be treated with lipoprotein apheresis (LA) in high-risk patients. We measured serum PCSK9 levels in patients eligible for the extracorporeal treatment: in 40 patients (Cohort I) who were treated with different systems before and after apheresis sessions and in the intervals between sessions. 10 patients (Cohort II) who were eligible but did not start LA yet served as controls. Patients' baseline serum PCSK9 levels were elevated relative to healthy volunteers and LA sessions acutely reduced the mean PCSK9 concentrations by 51%. Comparison of the effectiveness of the different LA methods demonstrated the DSA and HELP were more effective than the DALI system. After 24 h PCSK9 levels had returned to baseline compared to 8 days for the LDL-C concentrations to return to its pre-apheresis levels. In Cohort II baseline PCSK9 levels were similar to those in Cohort I. The acute reductions of PCSK9 by apheresis may be beneficial with respect to increasing the effectiveness of lipid-lowering drugs and with respect to an anti-atherosclerotic effect. In the future, antagonists to PCSK9 will probably be combined with or possibly replace LA in patients with a very high cardiovascular risk.


Effect of diltiazem on myocardial infarct size estimated by enzyme release, serial thallium-201 single-photon emission computed tomography and radionuclide angiography.
Diltiazem is a calcium antagonist with demonstrated experimental cardioprotective effects. Its effects on myocardial infarct size were studied in 34 patients admitted within 6 hours after the first symptoms of acute myocardial infarction. These patients were randomized, double-blind to placebo or diltiazem (10-mg intravenous bolus followed by 15 mg/hr intravenous infusion during 72 hours, followed by 4 X 60 mg during 21 days). Myocardial infarct size was assessed by plasma creatine kinase and creatine kinase-MB indexes, perfusion defect scores using single-photon emission computed tomography with thallium-201 and left ventricular ejection fraction measured by radionuclide angiography. Tomographic and angiographic scanning was performed serially before randomization, after 48 hours and 21 days later. Groups were comparable in terms of age, sex, inclusion time and baseline infarct location and size. Results showed no difference in creatine kinase and creatine kinase-MB data between controls and treated patients, a significant decrease in the perfusion defect scores in the diltiazem group (+0.1 +/- 3.0 placebo vs -2.2 +/- 1.9 diltiazem, p less than 0.02) and a better ejection fraction recovery in the diltiazem group (-4.2 +/- 7.4 placebo vs +7.7 +/- 11.2 diltiazem, p less than 0.05). Myocardial infarct size estimates from perfusion defect scores and enzyme data were closely correlated. These preliminary results suggest that diltiazem may reduce ischemic injury in acute myocardial infarction.(ABSTRACT TRUNCATED AT 250 WORDS)