Effects of weight change on taste function; a systematic review.
The aim of this review is to evaluate the relationship between weight status and taste perception and preference of sweet, salt, fat, bitter, and sour through reviewing observational and interventional studies with objective methods. A comprehensive literature search was performed in 6 online databases of PubMed, Scopus, Web of Science, Cochrane, Embase, and Google Scholar up to October 2021. The following keywords were used in the search strategy: (Taste OR "Taste Perception" OR "Taste Threshold" OR "Taste preference" OR "Taste sensitivity" OR "Taste changes") AND (weight OR "Weight gain" OR "weight loss" OR "weight change"). Most observational studies indicate that four taste sensitivities or perceptions (especially sweet and salt taste perception) are lower in subjects with overweight and obesity. The longitudinal studies reported that sweet and fat preference is increased along with weight gain in adults. It is concluded that taste perceptions are decreased in individuals with overweight and obesity, especially in men. Also, taste perception and preference change after weight loss but not significantly. It is suggested that the results of the interventional studies are not conclusive and need further studies with the same and standard design adjusting cofounding variables including genetic, gender, age and food condition of subjects.


Loss of FOXO1 promotes gastric tumour growth and metastasis through upregulation of human epidermal growth factor receptor 2/neu expression.
The biological significance of FOXO1, a member of the forkhead box O transcription factor family, in gastric cancer (GC) remains unclear. The present study provides direct evidence of the role of FOXO1 in tumour growth and metastasis of GC in relation to human epidermal growth factor receptor 2 (HER2). The expressions of FOXO1 and HER2 were modulated in GC cell lines (SNU-638, MKN45, SNU-216 and NCI-N87) by stable transfections. The effects of transfection on GC phenotypes were evaluated in vitro and in animal models. In addition, the relationship between FOXO1 and HER2 was analysed using GC clinical specimens, cell lines and xenografts. FOXO1 silencing in GC cells increased colony formation and mesenchymal transition in vitro, as well as tumour growth and metastasis in nude mice, whereas HER2 silencing induced the opposite results.. Furthermore, an inverse relationship between FOXO1 and HER2 was found in clinical specimens of GC, GC cells and GC xenograft tumours. Although a negative crosstalk between these two molecules was shown, double knockdown of both FOXO1 and HER2 in GC cells revealed that HER2 silencing reversed the FOXO1 shRNA-induced migration and invasion even without the FOXO1 restoration. Our results indicate that loss of FOXO1 promotes GC growth and metastasis by upregulating HER2 expression and that the HER2 expression is more critical to the induction of GC cell metastasis. The present study provides evidence that the FOXO1/HER2 pathway may regulate GC progression in a subgroup of GC patients.


A comparative evaluation of nutritional properties, antioxidant capacity and physical characteristics of cabbage (Brassica oleracea var. Capitate var L.) subjected to different drying methods.
Cabbag has been considered as one of attractive raw materials for dehydrated vegetables owing to high nutrient value. However, information about how drying methods affect the physicochemical properties of cabbage is limited. In present study, segmented cabbages were subjected to freeze drying (FD), hot air drying (HAD), microwave vacuum drying (MVD), vacuum drying (VD), MVD combined with HAD (MVD + HAD) and VD (MVD + VD). Dehydrated cabbages were compared in terms of nutritional components, antioxidant activity, microstructure, texture and so on. Results demonstrated that HAD conducted a worse effect, with loss rate of nutritional components >45%. However, two combined methods performed higher retention for nutritional compositions, better antioxidant activity and lower energy consumption than individual HAD. Furthermore, MVD + HAD products exhibited higher rehydration capacity and more uniform honeycomb porous structure but lower hardness compared to HAD. This conclusion would be fundamental for choosing more appropriate drying methods to obtain desirable quality.


Development of a multiplex PCR assay for the simultaneous and rapid detection of six pathogenic bacteria in poultry.
Escherichia coli, Pasteurella multocida, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella spp. and Staphylococcus aureus are six bacterial pathogens of avian. However, these pathogens may cause many similar pathological changes, resulting in clinical isolates that are difficult to quickly and simultaneously detect and identify. Here, a multiplex polymerase chain reaction (m-PCR) assay is reported to rapidly identify targets genes (phoA, KMT1, ureR, toxA, invA, and nuc) of these six pathogens in clinical samples. Six pairs of specific primers were designed. The optimal reaction conditions, specificity, and sensitivity of the m-PCR assay were investigated. The results showed that betaine remarkably improved amplification of the target genes. Specific test results showed that all six pathogens were detected by the proposed m-PCR protocol without cross-amplification with viruses or parasites. Sensitivity test results showed that the m-PCR system could amplify the six target genes from bacterial genomes or cultures with template amounts of 500 pg or 2.8-8.6 × 103 colony forming units, respectively. Furthermore, the six bacterial pathogens isolated from the infected tissue samples were successfully identified. The proposed m-PCR assay is a useful tool to monitor and diagnose bacterial infection in birds with high specificity, sensitivity and throughput.


Prenatal exposure to hexachlorobenzene (HCB) and reproductive effects in a multicentre birth cohort in Spain.
To investigate the possible association between birth size or gestational length and maternal serum concentrations of hexachlorobenzene (HCB) in a population exposed to background levels. A total of 1568 mother-child pairs recruited in three Spanish areas (INMA Project) from 2004 to 2008 participated in the study. Multivariate analysis was performed between birth weight and length, weeks of gestation, preterm birth or small for gestational age and HCB concentrations in maternal serum. The median concentration of HCB was 45.45 ng/g lipids. No association was found between HCB exposure levels and birth weight (β: -50.42 [-109.88; 9.04]), birth length (β: -0.07 [-0.32; 0.18]), gestation age (HR: 1.07 [0.94; 1.22]), small for gestational age (OR: 0.95 [0.56; 1.61]) and preterm birth (OR: 0.60 [0.29; 1.28]). Results remain similar after adjustment for other organochlorines. Our findings support the idea that exposure to low levels of HCB does not affect the intrauterine growth nor the duration of gestation.


Oxidative status and spermatozoa kinetics of rooster semen in citrus juice-based diluent.
Poultry spermatozoa are prone to oxidative damage and adversely affect their fertility. Bioactive constituents of citrus fruit confer antioxidant enrichment on its juice and are candidates to combat oxidative load in poultry semen. Computer-assisted semen analyser and oxidative status were used to evaluate the potency of two tropical citrus varieties (sweet orange and tangerine) as natural diluents for rooster semen. Fresh and ripe sweet orange and tangerine fruits were obtained and processed into juices using a standard protocol and included in dextrose saline at 0%, 10%, 20%, 30%, 40%, 50% and 60% as semen diluent. Semen pool from 30 breeder roosters of 35-40 weeks of age was allotted randomly in triplicates to the different fruit juice-dextrose at 1:2 dilution rate and evaluated for 5 h at room temperature. Diluted semen, according to treatments, was evaluated for sperm kinetics using a computer-assisted sperm analyser, and seminal plasma was assayed for lipid peroxidation and total antioxidant activity. The result obtained shows that tangerine and sweet orange juice inclusion significantly (p < 0.05) enhance progressive spermatozoa motility and semen kinetics compared favourably with undiluted semen. The diluents had a proportionate increase in antioxidant activity with juice inclusion at 0 h and the antioxidant activity of 40%, 50% and 60% fruit juice-based diluents was higher than undiluted semen at 5 h. There was a reduced lipid peroxidation in juice inclusive diluents and increase lipid peroxidation rate in undiluted semen and 0% juice inclusion. In conclusion, tangerine and sweet orange juice inhibit lipid peroxidation in rooster semen and enhance progressive spermatozoa motility and maintain rooster semen kinetics.


Low vitamin D and high cholesterol facilitate oral carcinogenesis in 4NQO-induced rat models via regulating glycolysis.
Diets and nutritional habits are critical during carcinogenic processes, where a diet poor in fruits and vegetables and rich in meat and other foods of animal origin facilitates carcinogenesis. In this study, we aimed at investigating the possible involvement of vitamin D deficiency (VDD) and high cholesterol (HC) together in oral squamous cell carcinoma (OSCC) through modulating glycolysis. We compared total cholesterol, LDL, HDL, triglycerides, LDH, and vitamin D levels of OSCC patients and control individuals. We used GEO datasets for gene set enrichment and 4-nitroquinoline-1-oxide induced in vivo oral carcinogenesis models to investigate contribution of VDD and HC during carcinogenesis via possible modulation of glycolysis. We found that VDD and HC co-exist in OSCC patients, and deregulation of cholesterol and vitamin D levels results in enrichment of genes related to glycolysis. We, then, demonstrated that VDD and HC on their own and together facilitated the formation of larger tumors in 4NQO-induced in vivo cancer models, which are suppressed by glycolysis inhibition. We reported collaborative contribution of HC and VDD during oral carcinogenesis, which is mainly carried out via altering energy metabolism in tumor cells.


Novel hit of DPP-4Is as promising antihyperglycemic agents with dual antioxidant/anti-inflammatory effects for type 2 diabetes with/without COVID-19.
DPP-4Is are well recognized therapy for type 2 diabetes. In spite of sharing a common mode of action, the chemical diversity among members of DPP-4Is raised the question whether structural differences may result in distinguished activities. DPP-4Is were recently explored as drug repurposing means for treatment of SARS-CoV-2 due to the urgent need for small molecule drugs for controlling infections. The use of DPP-4Is was not correlated with adverse COVID-19-related consequences among patients with type 2 diabetes. Inspired by these reasons and the importance of pyrimidinone ring as DPP-4I with both antioxidant and anti-inflammatory activities, we succeeded to prepare some novel pyrimidinone and thio-pyrimidinone derivatives, which were then screened for their antidiabetic activity and DPP-4 inhibition. In addition, their anti-inflammatory effect on LPS-stimulated RAW 264.7 cells were evaluated. Furthermore, their antioxidant activities were also tested.


N-3 fatty acids in the Mediterranean diet.
Fats in the diet of countries in the Mediterranean basin are typically represented by olive oil, but the high consumptions of vegetables and to some extent also of fish result in appreciable intakes of n-3 fatty acids. In fact, various plant foods are relatively rich in the 18 carbon n-3 fatty acid, alpha linolenic acid, ALA, while the generally moderate consumption of fish, except for certain communities living close to the sea, contributes to the intake of the long-chain n-3. Although the amounts of fats in ALA-containing plant foods are low, the relatively high concentrations of this fatty acid and the large size of the portions consumed allow to reach appreciable doses of ALA, an n-3 fatty acid that has been shown to exert favourable effects on various relevant factors in cardiovascular protection. In addition, consumption of relatively small amounts of certain typical dry fruit components of the diet such as walnuts, provides a sizable supply of ALA that is also rather efficiently converted to the ALA derivative eicosapentaenoic acid (EPA). Additional rather typical wild food components of the diet in certain countries, i.e. snails and frogs, are also appreciable sources of ALA. It appears thus that the consumption of typical Mediterranean foods provides relevant intakes of n-3 fatty acids, especially ALA, that appears to be efficiently absorbed and also transformed at least to the long-chain derivative EPA.


Positive link between variant Toll-like receptor 4 (Asp299Gly and Thr399Ile) and colorectal cancer patients with advanced stage and lymph node metastasis.
Toll-like receptors (TLRs) are considered as major endotoxin-signaling receptor and as crucial sensors of innate immunity. TLRs recognize pathogen-associated molecular patterns; induce effectors genes involving inflammatory cytokines and therefore initiation of adaptative immune responses against pathogens. Recently, it has been shown that TLRs are involved in tumor progression. In fact, increased level of TLR4 is associated with progression of colon malignancies. Even, TLR4 polymorphism has been shown associated with susceptibility to have colorectal cancer. Our study aimed to investigate an association between TLR4 Asp299Gly (D299G) and Thr399Ile (T399I) polymorphisms in Tunisian patients with colorectal cancer. Using a primer extension method (SNaPshot), we genotyped two variants of TLR4 D299G and T399I in 100 patients with colorectal cancer and 140 healthy controls in Tunisian population. Interesting, we noted a significant association between T399I polymorphism and tumor differentiation (p = 0.027) and tumor architecture (p = 0.02) in colorectal cancer (CRC) patients. We also showed a significant association of D299G with an increased risk of advanced stage (p = 0.03). Finally, we observed a positive link between D299G and T399I polymorphisms and CRC patients with lymph node (p = 0.00024; p = 0.0005, respectively) and metastasis (p = 0.001; p = 0.002, respectively). However, we found no evidence to support a significant association between TLR4 D299G and T399I polymorphisms and colorectal cancer susceptibility. Our findings suggest that TLR4 D299G and T399I polymorphisms are significantly associated with clinical features variables. TLR4 polymorphisms may serve as biomarker of disease progression. Therefore, our results need confirmation in even larger studies.


Immunohistochemical staining with EGFR mutation-specific antibodies: high specificity as a diagnostic marker for lung adenocarcinoma.
We previously demonstrated a high specificity of immunohistochemistry using epidermal growth factor receptor (EGFR) mutation-specific antibodies in lung adenocarcinoma and correlation with EGFR mutation analysis. In this study, we assessed EGFR mutation status by immunohistochemistry in a variety of extrapulmonary malignancies, especially those that frequently show EGFR overexpression. Tissue microarrays containing triplicate cores of breast carcinomas (n=300), colorectal carcinomas (n=65), pancreatic adenocarcinoma (n=145), and uterine carcinosarcoma or malignant mixed müllerian tumors (n=25) were included in the study. Tissue microarray of lung adenocarcinoma with known EGFR mutation status was used as reference. Immunohistochemistry was performed using antibodies specific for the E746-A750del and L858R mutations. In pulmonary adenocarcinoma, a staining intensity of 2+ or 3+ correlates with mutation status and is therefore considered as positive. Out of 300 breast carcinomas, 293 (98%) scored 0, 5 (2%) had 1+ staining, 2 (1%) were 2+ for the L858R antibody. All breast carcinomas scored 0 with the E746-A750 antibody. All the colorectal, pancreatic carcinomas and malignant mixed müllerian tumors were negative (0) for both antibodies. Molecular analysis of the breast carcinomas that scored 2+ for L858R showed no mutation. Our results show that EGFR mutation-specific antibodies could be an additional tool distinguishing primary versus metastatic carcinomas in the lung. False-positivity can be seen in breast carcinoma but is extremely rare (1%).


EPR spectroscopy studies on the structural transition of nitrosyl hemoglobin in the arterial-venous cycle of DEANO-treated rats as it relates to the proposed nitrosyl hemoglobin/nitrosothiol hemoglobin exchange.
In vivo studies show a dynamic cycle in which alpha-nitrosylated hemoglobin is mainly in the relaxed state in arterial blood of rats treated with 2-(N,N-diethylamino)-diazenolate-2-oxide, but converts mainly to the tense state during the arterial-venous transit. A detailed analysis shows that different electron paramagnetic resonance spectra recorded for alpha-nitrosyl hemoglobin in arterial and venous blood at 77 K originate only from a different ratio between 5- and 6-coordinate heme without any change in the concentration of nitrosyl hemoglobin. In venous blood, the five- and six-coordination equilibrium of the alpha-nitrosyl heme is shifted in favor of the 5-coordinate state (58% venous vs. 20% arterial). These results are not consistent with the recently proposed exchange of nitrosyl heme with the beta-93 nitrosothiol group of hemoglobin during the arterial-venous cycle.


Selective evolutionary pressure from the tissue microenvironment drives tumor progression.
Cells grow within defined environmental niches and are subject to microenvironmental control. Outside of their niche, the environment is hostile, the normal cells lack appropriate survival signals which leads to anoikis. During tumor development and progression, malignant cells must escape the local tissue control and resist anoikis. The inherent genetic instability of tumor cells makes their phenotype very plastic, which changes under continuous environmental selection pressure. In this way the microenvironment drives the somatic evolution of the tumor. In the current review, we assess how this environmental selection pressure fits into the classical scheme of tumor progression.


Degeneration and regeneration of olfactory cells induced by ZnSO4 and other chemicals.
The necrotic effect of various salt solutions was tested on the catfish olfactory mucosa. Only zinc cations were able to induce an extensive degeneration of the olfactory cells. Two different modes of irrigation of the mucosa with zinc sulfate were investigated. (1) The olfactory cavity is flushed with the chemical for not more than a few seconds. At concentrations above 30 mM, the resulting damage is very reproducible, largely concentration independent and almost completely specific for the olfactory receptor cells. The non-sensory respiratory cells are unaffected, the sustentacular cells surrounding the receptor cells are affected mainly by a loss of microvilli. The olfactory receptor cells, on the contrary, start to degenerate within a few hours and by day 4 only 20% of the original receptor population remains. Division of the mucosal basal cells increases during days 3 and 4 on and day 6 olfactory receptor cells reach the bare surface of the lamella. After day 7, the receptor population reaches a level of more than 80% of its original value. Because of the absence of sustentacular processes covering the olfactory cell's knobs on day 6, it has been possible to confirm that each of the two types of olfactory receptor cells previously characterized are concentrated on each half of the mucosa (2) The salt is maintained in contact with the tissue for several days. After this treatment most of the lamellae are irreversibly destroyed, some regeneration occurs in limited areas of the mucosa. In these small areas, indifferent respiratory cells reappear first between 20 and 35 days. It is only when the structure of the olfactory tissue is completely reorganized that the new receptor cells reappear between days 45 and 55. Regeneration is not completed before 60-65 days.


The dangers of the rabbit hole: Reflections on social media as a portal into a distorted world of edited bodies and eating disorder risk and the role of algorithms.
The relationship between social media usage and body image has been well-established in the literature; however, social media companies' use of algorithms may intensify this association, as algorithms provide viewers with personalized content that is often more extreme, less monitored, and designed to keep users engaged for longer periods of time. This article details the recent media coverage of algorithms, revelations by former social media employees regarding the problematic usage of algorithms, and revelations that social media companies are aware of the harm posed by their implementation of algorithms, particularly for young, vulnerable users. We provide recommendations for influencers, educators, researchers, clinicians, parents, and users, and conclude that it is ultimately the responsibility of the social media corporations to protect and enhance the well-being of their users.


Prosecutors' reflections on sexually abused preschoolers and their ability to stand trial.
Child sexual abuse (CSA) cases are notoriously difficult to investigate, and less than 10% of cases are prosecuted. We aimed to investigate prosecutors' experiences of preparing for and prosecuting suspected CSA cases with preschool aged victims. Nine specialized child prosecutors (6 women, 3 men) took part either in individual interviews or in focus groups on this subject. The transcripts were analyzed thematically. The prosecutors said that children's testimony was sometimes held to an adult standard and that child complainants who expressed emotion could be perceived as more credible than their less expressive counterparts. CSA victims were identified as vulnerable victims who had difficulty telling their stories. Some of the interviewers were described as lacking in the ability to approach these children. The results imply that the reliability and credibility of sexually abused preschoolers and their testimony might be influenced by a number of verbal and non-verbal factors and that there are several obstacles preventing prosecutors from prosecuting these cases.


Dismantling the tumoral cloak of self-protection.
Tumors protect themselves from immune clearance by promoting extramedullary hematopoiesis. A new study in PLOS Biology provides insights into the mechanisms underlying this process, which may hold the key to disrupting generation of the immunosuppressive tumor microenvironment.


High resolution ultrasonic neural modulation observed via in vivo two-photon calcium imaging.
Neural modulation plays a major role in delineating the circuit mechanisms and serves as the cornerstone of neural interface technologies. Among the various modulation mechanisms, ultrasound enables noninvasive label-free deep access to mammalian brain tissue. To date, most if not all ultrasonic neural modulation implementations are based on ∼1 MHz carrier frequency. The long acoustic wavelength results in a spatially coarse modulation zone, often spanning over multiple function regions. The modulation of one function region is inevitably linked with the modulation of its neighboring regions. Moreover, the lack of in vivo cellular resolution cell-type-specific recording capabilities in most studies prevents the revealing of the genuine cellular response to ultrasound. To significantly increase the spatial resolution, we explored the application of high-frequency ultrasound. To investigate the neuronal response at cellular resolutions, we developed a dual-modality system combining in vivo two-photon calcium imaging and focused ultrasound modulation. The studies show that the ∼30 MHz ultrasound can suppress the neuronal activity in awake mice at 100-μm scale spatial resolutions, paving the way for high-resolution ultrasonic neural modulation. The dual-modality in vivo system validated through this study will serve as a general platform for studying the dynamics of various cell types in response to ultrasound.


Incidence of ventricular shunt placement for hydrocephalus with clipping versus coiling for ruptured and unruptured cerebral aneurysms in the Nationwide Inpatient Sample database: 2002 to 2007.
Few studies have compared the incidence of ventricular shunt placement for hydrocephalus after clipping versus coiling of cerebral aneurysms. The Nationwide Inpatient Sample (NIS) database was used to compare, on a national level, the incidence of ventricular shunt placement after clipping versus coiling of ruptured and unruptured aneurysms. Hospitalizations for clipping and coiling of ruptured and unruptured aneurysms from 2002 to 2007 were collected from the NIS by cross-matching International Classification of Diseases-9 codes for diagnoses of subarachnoid hemorrhage or unruptured cerebral aneurysm with procedure codes for clipping or coiling. The incidence of ventricular shunt placement for hydrocephalus after clipping and coiling was compared using generalized linear models with generalized estimating equations (GEE) to adjust for patient- and hospital-specific factors and correlation between admissions. Of 10,899 ruptured aneurysm patients (6593 clipping, 4306 coiling), clipping had a similar incidence of ventricular shunt placement (9.3%) compared to coiling (10.5%) (odds ratio = 0.984; 95% confidence interval = 0.85, -1.14; P value = 0.833 after adjustment for patient-specific and hospital-specific factors). Likewise, of 9686 unruptured aneurysm patients (4483 clipping, 5203 coiling), clipping had similar incidence of ventricular shunt placement (0.4%) compared to coiling (0.5%) (odds ratio = 0.763; 95% confidence interval = 0.37, -1.58; P value = 0.465 after adjustment for patient-specific and hospital-specific factors). Predictors of shunt placement in ruptured aneurysm patients were age, comorbidity score, admission type, payer, and hospital aneurysm volume. Predictors of shunt placement in unruptured aneurysm patients were comorbidity score and admission type. In an observational study, clipping and coiling of ruptured and unruptured cerebral aneurysms are associated with similar incidences of ventricular shunt placement for hydrocephalus.


COVID-19 meningitis without pulmonary involvement with positive cerebrospinal fluid PCR.
Coronavirus disease 2019 (COVID-19) typically presents with respiratory illness ranging in severity. Neurological complications of the disease remain largely unknown. Herein, we discuss the case of a woman diagnosed with COVID-19 meningitis following two positive cerebrospinal fluid (CSF) RT-PCR assays, and highlight the importance of recognizing the neurological manifestations of the disease. The patient was a 49-year-old woman with a history of hypertension who presented with non-specific symptoms (fever, headache, malaise, nausea/vomiting). Chest computed tomography (CT) revealed a lack of pulmonary involvement and oropharyngeal/nasopharyngeal RT-PCR was negative for COVID-19. A lumbar puncture was performed on the third day of admission and the CSF analysis elucidated a viral pattern, but the CSF bacterial culture and RT-PCR assay for herpes simplex virus were both negative. Surprisingly, the CSF RT-PCR for COVID-19 was positive. The diagnosis of COVID-19 meningitis was made and the patient was treated solely with Kaletra® , with a second CSF analysis confirming our unique finding 1 week later. The patient's clinical characteristics improved progressively, and she was discharged in excellent general condition after 21 days. In contrast to what was originally believed, the SARS-CoV-2 can cause meningitis in isolation, perhaps by crossing the blood-brain barrier. Hence, it seems essential that physicians maintain a high index of suspicion for neurological involvement among COVID-19 patients, with early CSF analysis and brain imaging sometimes being indicated.


Electronic Health Record Embedded Strategies for Improving Care of Patients With Heart Failure.
A majority of clinical decisions use the electronic health record (EHR) and there is an unmet need to use its capability to help providers to make evidence-based decisions that improve care for heart failure patients. These electronic nudges are rooted in the human psychology of decision-making and often target specific cognitive biases. This review outlines the development of novel EHR nudges and specific lessons learned from each experience to inform the development of future interventions. There have been several randomized clinical trials examining the impact of EHR alerts on quality of care for heart failure patients. These interventions have targeted both clinicians and patients. There are features of each trial that inform best practices and future directions for EHR nudges. Recent clinical trials have demonstrated that some EHR alerts can improve care for heart failure patients. These trials utilized default options, involved clinicians in the alert design process, provided actionable recommendations, and aimed to minimize disruptions to typical workflow. Alerts aimed at improving care should be examined in a randomized fashion in order to evaluate their impact on clinician satisfaction and patient care.


Stevens-Johnson syndrome and toxic epidermal necrolysis in sub-Saharan Africa: a multicentric study in four countries.
The purpose of this study was to document the clinical profile, etiologies, and outcomes of Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) in hospitals in four sub-Saharan African countries. A retrospective study on cases of SJS/TEN treated in dermatology departments and/or intensive care units in four sub-Saharan African countries (Benin, Burkina Faso, Central African Republic, and Togo) from 2000 to 2010. The study focuses on variables such as age, sex, type of SJS/TEN, epidermal detachment of the skin surface, HIV status, drug(s) involved, and outcomes (death and sequelae). This study identified 177 cases of SJS/TEN from 2000 to 2010: 129 with SJS; 37 TEN; and 11 overlapping SJS/TEN. The average age of patients was 32.3 ± 15.4 years, and the sex ratio (M/F) was 0.6. HIV serology was positive in 69 (54.8%) of the 126 patients tested. Antibacterial sulfonamides (38.4%) were the most commonly used drugs followed by nevirapine (19.8%) and tuberculosis drugs (5.6%). We recorded 22 deaths (i.e. six cases of SJS, 15 of TEN, and one of overlapping SJS/TEN). Of the 22 patients who died, 16 were infected with HIV; among them, seven had an opportunistic infection (four cases of cerebral toxoplasmosis and three of pulmonary tuberculosis). Twenty-seven cases of sequelae were noted with a large part of eye complications. This study has highlighted: (i) the high proportion of patients infected with HIV among patients who had SJS/TEN in sub-Saharan Africa; (ii) the high frequency of antiretroviral drugs as new SJS/TEN causes in sub-Saharan Africa; and (iii) the impact of HIV infection on morbidity and mortality of these affections.


Benefits of applying organic amendments from recycled wastes for fungal community growth in restored soils of a limestone quarry in a semiarid environment.
Applying organic amendments to recover physical, chemical, and biological qualities of soil may enable recovery of soils degraded by mining in semiarid climates. This study's aim was to investigate the development and changes in the composition of fungal communities in restored soils with five different types of organic amendments (two types of vegetable compost and sewage sludge compost, and a mixture of both) compared with unamended soils and surrounding natural soils and to examine the relationships between the fungal taxa, the new physico-chemical and biological soil properties of technosoils after 18 months of restoration, and natural soils. Restoration improved soil quality and fungal diversity, placing these soils in an intermediate position between unrestored soils (with no fungi present) and undisturbed reference soils, which were the most fungal diverse. Sewage-treated soils and their mixtures showed high nitrogen and carbohydrate content as well as high basal respiration and fatty acid content, suggesting that they provided readily biodegradable organic matter. In contrast, greenhouse compost-treated soils showed high total organic carbon and polyphenol content, whereas garden compost-treated soils showed intermediate values. The biological soil properties of both composts showed were similar to those of the reference soils, suggesting that composts contained more resilient organic matter. Organic amendments of dissimilar origin caused significantly different fungal soil communities at the genus level among the restored soils. Results indicated that soil pH, electrical conductivity, total nitrogen content, soil basal respiration, fungi/bacteria-PLFA ratio, and dehydrogenase and β-glucosidase activities, together with Pearson's correlations, revealed that these properties and nutrient content (total organic carbon, C/N ratio, carbohydrates, and polyphenols) influenced 40 soil fungal taxa. Therefore, the organic amendments led to changes in soil properties that favoured plant cover by promoting the soil fungal community growth beneficial to the carbon cycle and symbiotic with plants.


Maturation of the auditory brain stem response in rhesus monkeys (Macaca mulatta).
To document the maturational changes in the auditory brain stem response (ABR) of the rhesus monkey, longitudinal and cross-sectional data were collected on a panel of 31 animals during the first year of life. Each ABR test consisted of averaging 2048 responses to click stimuli delivered at a rate of 21 clicks/sec and elicited at 60, 40, 20 and 10 dB HL intensities in accordance with procedures described previously. The morphology of the ABR wave form recorded at birth was similar to that at 12 months of age and the latencies of the component waves decreased exponentially over this period. The magnitude of decrease in latency was greatest for wave IV and least for wave I. The regression lines for the log of the latencies of 3 waves on age were computed for 3 animals followed longitudinally and for the cross-sectional data set. The slopes of these regression lines were different among animals, indicating differing rates of maturation. However, these slopes were identical for the component waves of any one animal, suggesting that the rate of maturation was constant over the auditory pathway. These changes were similar in pattern to those reported for human data and suggest that the rhesus monkey may serve as an appropriate model of auditory development in man.


Primary aortic intimal sarcoma.
Aortic intimal angiosarcoma is extremely rare, and the prognosis of patients with a tumor is unfavorable even if they have undergone surgery. We treated a patient with primary intimal angiosarcoma of the aortic arch who underwent an operation. The tumor originated from the inner wall of the aortic arch on the lesser curve. In order to remove the tumor completely, the entire aortic arch from the ascending to the middle of descending aorta was resected through an L-shape skin incision. On histologic examination, an undifferentiated intimal sarcoma was diagnosed. It grew into the aortic lumen while spreading along the aortic intima, focally infiltrating the media. The postoperative course was uneventful. Postoperative CT performed at 6, 12 and 18 months after surgery showed no local recurrence or metastasis. According to some reports endoarterectomy has been performed to treat this type of tumor, since the malignant cells are thought to be limited to the luminal surface. However, we favor aortic resections and graft interpositions rather than an endoarterectomy because the tumor could have invaded the media.


Clinical Update for Physical Therapists: Coagulopathy and COVID-19.
Physical therapists have a unique role in both prevention of venous thromboembolism (VTE) through the promotion of early mobility and physical activity and diagnosis through discovery of signs and symptoms of VTE. This Perspective updates clinicians on the latest information regarding pathophysiology of coagulopathy associated with COVID-19 and applies VTE clinical practice guidelines to COVID-19 in order to provide guidance on physical therapist management.


Sequential studies on the role of mitoxantrone in the treatment of acute leukemia.
Mitoxantrone (Novantrone; 1, 4-dihydroxy-5, 8-bis [[2-[(2-hydroxyethyl) amino]ethyl]amino-] 9, 10 anthracenedione dihydrochloride (NSC 301739] is a synthetic anthracenedione with intercalating properties. Activity has been shown in preclinical studies in mice bearing intraperitoneal P388 and L1210 leukaemias, ADJ-Pc6 plasmacytoma and a variety of solid tumours. In a phase I/II collaborative study fourteen consecutive patients with relapsed or primarily refractory acute leukaemia received a single infusion of mitoxantrone (20-32 mg/m2) at fourteen-day intervals. Antileukaemic activity was seen but there were no complete remissions and toxicity was minimal. Mitoxantrone was subsequently given in a five-day schedule at a dose of 10mg/m2 daily to twenty-one patients with relapsed or refractory acute leukaemia or chronic myeloid leukaemia in blast crisis (CML-BC). Four of five patients in first relapse of acute non-lymphoblastic leukaemia (ANLL) achieved a complete remission (CR). The overall response rate (CR + partial remission (PR] was 48%. In an ongoing phase III study the same (5-day) mitoxantrone treatment has been given in conjunction with a 7-day continuous infusion of cytosine arabinoside (Ara-C) in a kinetically designed schedule based upon the preclinical studies of the Mount Sinai group.


The pediatrician's role in atherosclerosis prevention.
Prevention of atherosclerosis, a major cause of illness and death in our society, is a pediatric responsibility in 1988 and beyond. Significant cardiovascular risk factors are identifiable and alterable in children and adolescents, and are associated with arterial lesions. Cost-effective atherosclerosis prevention can be achieved by identification and treatment of high-risk children and adolescents. The pediatrician should record the family history of all patients at 2 years of age, with periodic updates. The serum cholesterol concentration can then be measured in the office if there is a commitment to quality control; otherwise, a monitored commercial laboratory must be used. It is not unreasonable to measure serum cholesterol concentrations once in all school-age patients. The majority of high-risk patients will respond to nutritional intervention and will not require pharmacotherapy. Until the efficacy and safety of the therapeutic diet are proved beyond doubt, its use should be limited to high-risk patients under pediatric supervision. Studies documenting the growth and development of children receiving dietary therapy must be a major priority of pediatric nutrition. Until that is accomplished, although some pediatricians may prefer to wait, most will use the extensive scientific evidence at hand and their clinical judgment to identify and treat their high-risk patients.


On the possible effects of homeostatic shifts in human embryonic development.
Possible etiological factors of congenital malformations as well as of human trisomies are considered in the framework of the repressor hypothesis. In this approach gene expression is envisaged from the point of view of the functional variations of the total activation energy for normal gene expression in homeostatic equilibrium. We restrict our attention to variations of the total activation energy under the effect of temperature gradients. We discuss the evidence that hyperthermia may be an etiological factor for trisomies in humans.


Molecular mechanisms of developmental toxicities of azoxystrobin and pyraclostrobin toward zebrafish (Danio rerio) embryos: Visualization of abnormal development using two transgenic lines.
Azoxystrobin (AZ) and pyraclostrobin (PY) are strobilurin fungicides that inhibit fungal mitochondrial respiration. In this study, a representative model, zebrafish (Danio rerio), was used as a test species for acute and developmental toxicity. Survival and malformation rates were observed only PY-treated embryos, with an LC50 value of 77.75 ppb accompanied by a dramatic decrease in hatching rate, while AZ did not show great mortality. Morphological changes were observed in PY-treated embryos with the occurrence of pericadial edema at 25 ppb. A delay in growth was observed after treatment with pyraclostrobin at 50 ppb. Use of genetically engineered Tg(cmlc:EGFP) allowed fluorescence observation during heart development. PY interfered with normal heart development via upregulation of the nppa gene responsible for the expression of natriuretic peptides. Heart function was dramatically reduced as indicated by reduced heart rates. Increased expression of the nppa gene was also seen in AZ-treated embryos. The expression level of cyp24a1 was also up-regulated, while ugt1a1 and sult1st6 were down-regulated after treatment of zebrafish embryos with AZ or PY. Overall, strobilurin fungicides might inhibit normal heart formation and function within the range of concentrations tested.


Altered promoter binding of the TATA box-binding factor induced by the transcriptional activation domain of VP16 and suppressed by TFIIA.
The acidic transcriptional activation domain of the Herpes simplex virus protein VP16 has been shown to bind directly to both the TATA box-binding factor TBP and the general initiation factor TFIIB. Using DNase I footprinting assays, we have shown here that the VP16 activation domain qualitatively alters binding of Saccharomyces cerevisiae TBP to a TATA sequence in DNA. The effect of VP16 on promoter binding by TBP was reduced by mutations in VP16 known to reduce transactivation and could not be overcome by increasing the amount of TBP used in the footprinting assays. However, the association of yeast TFIIA with TBP on the promoter reversed the VP16-mediated effect and restored normal binding of TBP to the promoter. We suggest that VP16 induces a conformational change in TBP which alters its binding to promoter DNA, and that this effect of VP16 is suppressed by TFIIA.


Epitopes recognized by antibodies to denatured core protein of hepatitis B virus.
Particulate and denatured core protein as well as e-antigen (HBe) of hepatitis B virus (HBV) differ in part immunologically but this has not been studied in sufficient detail. Therefore, in this study the B-cell immune response to native and denatured HBV core protein which both can exhibit HBe-specific epitopes was examined using a panel of mouse MABs and rabbit polyclonal antibodies to native and denatured core protein and polyclonal anti-HBe/anti-HBc antibodies from sera of infected patients. Epitope mapping was performed using a set of partially overlapping synthetic HBc peptides, carboxy-terminally truncated HBc proteins and various HBc fusion proteins. A major immunogenic region between amino acids 134-140 and two less immunogenic regions, one spanning amino acids 2-10 and one with three partially overlapping epitopes between amino acid positions 138 and 154, were defined by mouse MABs. Polyclonal rabbit antibodies to denatured HBc, woodchuck and ground squirrel hepatitis core proteins (WHc and GSHc) recognized similar epitopes but in addition occasionally region 61-85, and the latter was also recognized on particulate HBc. Two antigenic regions (amino acid positions 2-10 and 138-145) were found to be exposed on HBe from human serum, and were recognized by mouse anti-HBe but not by anti-HBc antibodies from sera of infected patients. This study demonstrates a more complex pattern of HBc and HBe epitopes than detected previously and provides tools to study conformational changes which may take place during HBc/HBe processing, transport and core particle assembly.


On the intracellular release mechanism of hydrophobic cargo and its relation to the biodegradation behavior of mesoporous silica nanocarriers.
The intracellular release mechanism of hydrophobic molecules from surface-functionalized mesoporous silica nanoparticles was studied in relation to the biodegradation behavior of the nanocarrier, with the purpose of determining the dominant release mechanism for the studied drug delivery system. To be able to follow the real-time intracellular release, a hydrophobic fluorescent dye was used as model drug molecule. The in vitro release of the dye was investigated under varying conditions in terms of pH, polarity, protein and lipid content, presence of hydrophobic structures and ultimately, in live cancer cells. Results of investigating the drug delivery system show that the degradation and drug release mechanisms display a clear interdependency in simple aqueous solvents. In pure aqueous media, the cargo release was primarily dependent on the degradation of the nanocarrier, while in complex media, mimicking intracellular conditions, the physicochemical properties of the cargo molecule itself and its interaction with the carrier and/or surrounding media were found to be the main release-governing factors. Since the material degradation was retarded upon loading with hydrophobic guest molecules, the cargo could be efficiently delivered into live cancer cells and released intracellularly without pronounced premature release under extracellular conditions. From a rational design point of view, pinpointing the interdependency between these two processes can be of paramount importance considering future applications and fundamental understanding of the drug delivery system.


Is there a relationship between middle concha bullosa and ethmoid roof asymmetry?
The middle turbinate and ethmoid roof are intranasal structures and may have many anatomical variations. These structures, which serve as anatomical markers during functional sinus surgery, are important for preventing complications and performing a proper surgery. Knowledge of anatomical variations will increase surgical success and reduce complications. We aimed to investigate the presence of asymmetry in the ethmoidal roof and anatomical variation in patients with and without concha bullosa. In this study, the files of patients who underwent paranasal computed tomography between 2012 and 2018 were analyzed retrospectively. The patients were divided into two groups, as patients with and without concha bullosa. Differences between the two groups in terms of age, gender, septum deviation, ethmoid artery dehiscence, ethmoid roof asymmetry were examined. The 369 patients included in our study were divided into two groups; those with concha bullosa and those without concha bullosa. The mean age of the patients with concha bullosa was 36.1 ± 13.4 (min-max: 12-74) and the mean age of patients without concha bullosa was 37.5 ± 14.3 (min-max: 10-81). The ethmoid roof depths were compared between the two groups and a significant difference was observed (p < 0.001). The ethmoid roof depth was higher in the group with concha bullosa (p < 0.001). The results of our study indicate that the ethmoidal roof tends to be higher in patients with middle concha bullosa.


Effects of wheels and tires on high-strength lightweight wheelchair propulsion cost using a robotic wheelchair tester.
This study was designed to investigate the effect of wheel and tire selections on the propulsion characteristics of a high-strength lightweight manual wheelchair using robotic wheelchair propulsion. Four configurations were compared with differing combinations of drive wheel tires and casters, with the baseline reflecting the manufacturer configuration of a solid mag drive wheel and 8"×1" caster. The robotic wheelchair tester propelled the chair using pre-generated straight and curvilinear manoeuvres using repeatable and reliable cyclic torque profiles. Additionally, energy loss of the components was measured using coast-down deceleration tests to approximate the system-level rolling resistance of each configuration. Results indicate a significant decrease in propulsion cost, increased distance travelled and increased manoeuvrability across all configurations, with upgraded casters and tires. These results indicated that with better casters and drive wheel tires, the performance of high strength lightweight wheelchairs can be improved and better meet the mobility needs of users.Implications for rehabilitationWheel and tire selection can have a demonstrable impact on the propulsion efficiency of manual wheelchairsCoast-down test protocols can be used as a simple and cost-effective means of assessing representative energy losses across various surfacesWheelchair configurations can be optimized with proper knowledge of the main energetic loss contributions and the environments and contexts of use.


Successful surgical removal of a large arteriovenous malformation in a patient with hemophilia: case report.
This is the first reported case of the successful surgical removal of a large arteriovenous malformation (AVM) in a patient with hemophilia A. A 19-year-old male patient was admitted to our department with intracranial hemorrhage. He had previously been diagnosed with hemophilia A and a cerebral AVM. Carotid angiography revealed a large AVM in the right temporal and parietal lobes. The neurological and neuroradiological findings, especially those of single photon emission computed tomography, identified an area of devitalization around the lesion, which was thought to reduce the risk of new deficits resulting from surgical manipulation. We resected the AVM in conjunction with supplemental infusions of Factor VIII before, during, and after the operation. A slight cerebral hemorrhage on the 7th postoperative day was observed despite control with Factor VIII, but the patient was discharged without any new deficits. We evaluated and managed all problems of a patient with multiple complications and achieved a medical cure.


[Finite element analysis of the effects of two kinds of individual extralevator abdominalperineal excision on female pelvic floor].
To evaluate the effects of two kinds of individual extralevator abdominalperineal excision(ELAPE) on female pelvic floor biomechanics. Four kinds of finite element model were developed: intact models, ELAPE models and two kinds of individual ELAPE models. The maximal stress and stress distributions of each model under the same pressure were analyzed and compared. In the individual ELAPE models of dissection plane close to rectum on one side, non-levator ani tissue's maximal stress on levator ani partly reserved side and totally removed side were (3.114±0.129) and (4.856 ±0.128) MPa. The former were lower than the latter (P<0.01). They were both lower than those in ELAPE models which were (5.127±0.070) MPa (P<0.01), and were higher than those in intact model which were (1.963±0.061) MPa (P<0.01). In the individual ELAPE models of dissection plane close to pelvic on one side, non-levator ani tissue's maximal stress on both sides, which were (5.131±0.067) MPa, were similar to those in ELAPE models (P>0.05). ELAPE of dissection plane close to rectum on one side is able to reduce the stress of non-levator ani tissue on both sides, especially on levator ani partly reserved side; conversely, ELAPE of dissection plane close to pelvic on one side may not reduce the stress of non-levator ani tissue on either side.


Assessment of L-ascorbic acid requirement for prolonged survival in ODS rats and their susceptibility to urinary bladder carcinogenesis by N-butyl-N-(4-hydroxybutyl)nitrosamine.
Assessment of L-ascorbic acid requirement for prolonged survival in ODS (genotype: od/od) rats and their susceptibility to urinary bladder carcinogenesis by N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN) were examined. In ODS rats without L-ascorbic acid synthesizing ability, the 50 ppm dietary total ascorbic acid (TAA) was insufficient to survive for 4 weeks, the 250 ppm dietary TAA was sufficient to survive for 36 weeks. In examination of BBN treatment, ODS rats--although showing a lower availability of TAA than the heterozygotes (+/od) and normal (+/+) rats with L-ascorbic acid synthesizing ability--were equally susceptible to bladder carcinogenesis.


Is the walk ratio a window to the cerebellum in multiple sclerosis? A structural magnetic resonance imaging study.
Existing research studies have demonstrated a relationship between magnetic resonance imaging (MRI) neuroimaging measures and walking speed in people with multiple sclerosis (PwMS). However, to date there are no data as to the brain structures involved in gait coordination and control in PwMS. Therefore, the aim of our study was to investigate the association between walk ratio, an indicator of gait coordination, and related brain structures in PwMS. A brain MRI was performed by a 3.0-T MR scanner in conjunction with a volumetric analysis based on three-dimensional T1-weighted images. Regions of interest were volumes of the hippocampus, amygdala, putamen, caudate, pallidum, thalamus, cerebellum and the corpus callosum regions. Walking speed and walk ratio, defined as step length divided by step rate, was measured whilst walking on an electronic walkway. In all, 343 PwMS (41.1 ± 13.4 years, 69.1% female, median Expanded Disability Status Scale 2.5) were included in the study. A significant association was found between the left cerebellum volume and walk ratio after controlling for age, gender, total cranial volume and disability; R2 = 0.379, P = 0.002. A similar association was found between the right cerebellum volume and walk ratio, R2 = 0.364, P = 0.002. No correlations were observed between walk ratio and the thalamus, basal ganglia, hippocampus, amygdala and the corpus callosum volumes. No association was found between walking speed and all brain measures. The walk ratio should be considered when evaluating and assessing PwMS presenting with ataxia. Furthermore, it is also hypothesized that a low walk ratio indicates a lower cerebellum volume in the MS population.


The emerging role of peptides and lipids as antimicrobial epidermal barriers and modulators of local inflammation.
Skin is complex and comprised of distinct layers, each layer with unique architecture and immunologic functions. Cells within these layers produce differing amounts of antimicrobial peptides and lipids (sphingoid bases and sebaceous fatty acids) that limit colonization of commensal and opportunistic microorganisms. Furthermore, antimicrobial peptides and lipids have distinct, concentration-dependent ancillary innate and adaptive immune functions. At 0.1-2.0 μM, antimicrobial peptides induce cell migration and adaptive immune responses to coadministered antigens. At 2.0-6.0 μM, they induce cell proliferation and enhance wound healing. At 6.0-12.0 μM, they can regulate chemokine and cytokine production and at their highest concentrations of 15.0-30.0 μM, antimicrobial peptides can be cytotoxic. At 1-100 nM, lipids enhance cell migration induced by chemokines, suppress apoptosis, and optimize T cell cytotoxicity, and at 0.3-1.0 μM they inhibit cell migration and attenuate chemokine and pro-inflammatory cytokine responses. Recently, many antimicrobial peptides and lipids at 0.1-2.0 μM have been found to attenuate the production of chemokines and pro-inflammatory cytokines to microbial antigens. Together, both the antimicrobial and the anti-inflammatory activities of these peptides and lipids may serve to create a strong, overlapping immunologic barrier that not only controls the concentrations of cutaneous commensal flora but also the extent to which they induce a localized inflammatory response.


Multiple adverse experiences and child cognitive development.
During childhood and adolescence, children's social environments shape their cognitive development. Children exposed to multiple adversities in their social environment are more likely to have poorer cognitive outcomes. These findings have prompted interest among pediatric and public health communities to screen and connect youth to appropriate interventions that ameliorate the detrimental effects of adverse exposures. Such intervention efforts can be improved with a stronger conceptual understanding of the relationship between multiple adverse exposures and child cognitive development. This includes disentangling adverse exposures from other risk factors or underlying mechanisms, specifying mechanisms of action, and determining when adverse exposures are most detrimental. This review summarizes findings from the literature on each of these areas and proposes a conceptual model to guide further research and intervention.


A nationwide study of occurrence and exposure assessment of neonicotinoid insecticides and their metabolites in drinking water of China.
Neonicotinoid insecticides (NNIs) are the most extensively used insecticides worldwide, threatening ecosystem and human health. However, nationwide studies of NNIs and their metabolites in drinking water are limited. In order to characterize the contamination status of NNIs in drinking water throughout China, we collected 884 drinking water samples including 789 tap water and 95 groundwater samples from 32 provinces (covering seven regions of mainland China: south, central, east, north, northeast, northwest, and southwest) and Hong Kong. Ten NNIs and six of their main metabolites were determined in the water samples. The relative potency factor method was used to assess the cumulative concentrations of NNIs and their metabolites (imidacloprid-equivalent total NNIs, IMIeq) based on the chronic reference doses (cRfDs) of the NNIs or the toxic effects of the mataboilites. The IMIeq varied among the studied regions, with a median concentration of 24.5 ng/L and a maximum concentration of 8,622 ng/L. The predominant NNIs in drinking water were acetamiprid (ACE) and imidacloprid (IMI). Compared with tap water derived from groundwater, much higher concentrations of IMIeq and NNIs were found in tap water derived from surface water. Different concentrations and patterns of NNIs in drinking water were observed in different regions, provinces, and capital cities, mainly due to regional and provincial differences in crop types and volumes of pesticide usage. The concentrations of NNIs in the drinking water of provincial capitals and small/medium cities were higher than the concentrations in rural areas. The estimated daily intake (EDI) of IMIeq was at least two orders of magnitude lower than the cRfD of IMI, while the NNIs in 16 drinking water samples exceeded the acceptable value (100 ng/L) recommended by the European Union. This study provided a nationwide profile of the occurrence of NNIs and their metabolites in the drinking water of China and the associated potential cumulative human health risks, taking into account of the toxicity differences between NNIs and their metabolites.


Protamine alterations in human spermatozoa.
Protamines are the major nuclear proteins in sperm cells, having a crucial role in the correct packaging of the paternal DNA. The fact that protamine haploinsufficiency in mice resulted in abnormal chromatin packaging and male infertility suggested that the protamines could also be important candidates in explaining some of the idiopathic male infertility cases in humans. The first clinical studies focused on analyzing protamines at the protein level. Various studies have found the presence of an altered amount of protamines in some infertile patients, in contrast to the normal situation in fertile individuals where the two protamines, protamine 1 and protamine 2, are both present in approximately equal quantities. Subsequently, the protamine genes were the subject of various mutational genetic screening studies in search of variants that could be associated with deregulation in the protamine expression observed. The results of these protamine mutational studies showed that the presence of high penetrant mutations is a very rare cause of male infertility. However, some variants and some haplotypes described may behave as risk factors for male infertility. More recently, the presence of RNA in the mature sperm cell has also been investigated. The present chapter will introduce the basic aspects of protamine evolution and function and review the various articles published to date on the relationship between the protamines studied at the DNA, RNA, and protein levels and male infertility.


Men at risk of gonococcal urethritis: a case-control study in a Darwin sexual health clinic.
Male urethritis is primary sexually transmitted. Northern Territory (NT) has the highest rates of gonococcal infection in Australia and local guidelines recommend empiric treatment with azithromycin and ceftriaxone for all men presenting with urethritis. As gonococcal drug resistance is a growing concern, this study aims to improve empiric use of ceftriaxone through examining local patterns of male urethritis, comparing cases of gonococcal urethritis (GU) to controls with non-gonococcal urethritis (NGU). A retrospective study was undertaken of all men with symptomatic urethritis presenting to Darwin sexual health clinic from July 2015 to July 2016 and aetiology of urethritis in this population was described. Demographic, risk profile, and clinical features of GU cases were compared to NGU controls. Among n = 145 men, the most common organisms identified were Chlamydia trachomatis (23.4%, SE 3.5%) and Neisseria gonorrhoeae (17.2%, SE 3.1%). The main predictors of GU were any abnormalities on genital examination (aOR 10.4, 95% CI 2.1 to 50.8) and a history of urethral discharge (aOR 5.7, 95% CI 1.4 to 22.6). Aboriginal patients (aOR 3.0, 95% CI 0.9 to 9.6) and those over 30 years of age (aOR 1.4, 95% CI 0.3 to 7.0) were more likely to have GU in the unadjusted analysis, but not in the adjusted model. This is the first study looking at patterns of male urethritis in urban NT and the results support a move towards adopting national guidelines to use ceftriaxone for empiric management of syndromic urethritis only in high-risk patients. In addition to traditional demographic risk factors, clinical features remain an important component of risk stratification.


Epigenetic alchemy for cell fate conversion.
Recent progress in neural stem cell research shows that a number of extrinsic factors and intracellular mechanisms, including epigenetic modifications, are involved in the self-renewal of neural stem cells and in neuronal and glial differentiation. Remarkably, there is increasing evidence that the remodeling of chromatin structure and the alteration of epigenetic marks, including histone methylation and acetylation and DNA methylation, can cause committed cells to convert from one fate to another, and such converted cells are functional when transplanted in vivo. Thus, epigenetic research might generate the alchemy required to convert any non-neural stem cells into functional neural stem cells, which are few and difficult to extract from the adult central nervous system.


Returning a Shelter Dog: The Role of Owner Expectations and Dog Behavior.
Millions of animals are adopted from animal shelters in the United States each year, although some are returned post-adoption, which can decrease both the animals' chances of future adoptions and the owners' willingness to adopt again. In this study, we investigated the impact of adopter expectations for ownership and animal behavioral problems on post-adoptive dog returns at a large animal shelter in South Carolina. Between June-September 2021, 132 dog adopters completed a survey about their expectations for ownership through Qualtrics. Twenty-nine adopters returned their dogs to the shelter within three months of adoption, with a median length of ownership of eight days. Owners completed follow-up questionnaires about their perceptions of adoption and dog behavior at two days, two weeks, and four months post-adoption. Categorical principal component analysis revealed three factors pertaining to adopters' expectations for ownership. Independent t-tests showed returning owners had significantly higher expectations for dog behavior and health (t = -2.32, p = 0.02) and the human-dog bond compared with non-returning owners (t = -2.36, p = 0.02). Expectations for ownership responsibilities did not differ between the groups. Two-thirds of adopters experienced dog behavioral problems post-adoption, although training difficulty decreased significantly between two days and four months (F = 5.22, p = 0.01) and nonsocial fear decreased between two weeks and four months post-adoption (X2 = 10.17, p = 0.01). Shelters may benefit from utilizing adoption counselling to ensure adopters understand the potential for dog behavioral problems in the early stages of ownership and develop appropriate expectations for the human-dog relationship. Post-adoption behavioral support may also help some owners to overcome behavioral difficulties as their dogs adapt to the new environment.


Chronic liver diseases: From development to novel pharmacological therapies: IUPHAR Review 37.
Chronic liver diseases comprise a broad spectrum of burdensome diseases that still lack effective pharmacological therapies. Our research group focuses on fibrosis, which is a major precursor of liver cirrhosis. Fibrosis consists in a progressive disturbance of liver sinusoidal architecture characterised by connective tissue deposition as a reparative response to tissue injury. Multifactorial events and several types of cells participate in fibrosis initiation and progression, and the process still needs to be completely understood. The development of experimental models of liver fibrosis alongside the identification of critical factors progressing fibrosis to cirrhosis will facilitate the development of more effective therapeutic approaches for such condition. This review provides an overlook of the main process leading to hepatic fibrosis and therapeutic approaches that have emerged from a deep knowledge of the molecular regulation of fibrogenesis in the liver. LINKED ARTICLES: This article is part of a themed issue on Translational Advances in Fibrosis as a Therapeutic Target. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v180.22/issuetoc.


Histopathological and immunohistochemical (cytokeratins AE1/AE3) study of the parametrium of patients with early stage cervical cancer.
This study was undertaken in order to evaluate histopathological and immunohistochemical (cytokeratins AE1/AE3) characteristics of parametrial invasion in patients with early stage cervical cancer. Thirty patients with stage IB squamous cell carcinoma (SCC) of the cervix submitted to radical hysterectomy from November 2001 to September 2002 were prospectively studied. Histopathological studies were undertaken using tissue samples (n=60) taken from the parametrium, whose surgical margins were inked and the entire parametrial tissue was fixed in 10% buffered formalin and embedded in paraffin for further analysis using hematoxylin-eosin (H&E) staining. Specific patterns of parametrial involvement (continuous invasion, parametrial lymphatic vascular space invasion (LVSI) and/or parametrial lymph nodes' (PMLN) metastasis) were recorded. Parametrial samples, in which the histological examination showed no tumor involvement, were immunohistochemically assessed through monoclonal antibodies for cytokeratins AE1/AE3. Clinicopathological characteristics of the patients were also recorded. Patient's mean age was 49+/-10 years (27-73 years). Histopathological analysis (H&E) showed parametrium involvement in 12 patients (40%) of whom 11 (92%) presented parametrial LVSI, 9 (75%) continuous invasion and 4 (33%) PMLN metastasis. Micrometastasis was detected in 3/18 (17%) of the patients with histologically negative parametrium by H&E evaluation. Parametrial involvement detected by H&E was associated with tumor recurrence (p=0.009) and survival (p=0.025). This association was not correlated with the presence of parametrial micrometastasis (p=1.00 and 1.00, respectively). The process of parametrial spreading in patients with SCC of the cervix may develop several histopathological patterns, which are associated with clinicopathological features and prognosis. Our findings highlight the importance to ink the parametria, which is the only way to define the pattern of tumor spreading. The clinical significance of micrometastasis, detected in patients with histologically negative parametrium by H&E, remains to be clear.


Electrocardiographic Parameters in Indian Newborns.
To generate data of electrocardiogram (ECG) parameters according to gestational age in Indian newborns. An observational study was carried out over 7 months in neonatology unit of a tertiary care teaching hospital. Following auscultation, ECG parameters were recorded simultaneously in 12 leads, on third day of life, in hemodynamically stable neonates. Data from 364 babies were analyzed, keeping at least 30 records for each gestational age between 30 to 42 weeks. There was no difference in mean heart rate recorded through auscultation and ECG traces. The mean (SD) values recorded were: P wave duration 0.04 (0.01) s, P wave amplitude 1.3 (0.4) mm, T wave duration 0.07 (0.02) s, T wave amplitude 1.1 (0.6) mm, PR interval 0.09 (0.02) s, QRS duration 0.04 (0.01) s, QT interval 0.26 (0.02) s, QTc 0.4 (0.03) s and QRS axis 127 (22) degree. Gestation age-wise percentile charts of different ECG parameters were generated. These gestational age-wise percentile charts of different ECG parameters for Indian newborns can be used as reference for neonatal ECG.


Methylxanthines and ryanodine receptor channels.
Methylxanthines of either natural or synthetic origin have a number of interesting pharmacological features. Proposed mechanisms of methylxanthine-induced pharmacological effects include competitive antagonism of G-coupled adenosine A(1) and A(2A) receptors and inhibition of phosphodiesterases. A number of studies have indicated that methylxanthines also exert effects through alternative mechanisms, in particular via activation of sarcoplasmic reticulum or endoplasmic reticulum ryanodine receptor (RyR) channels. More specifically, RyR channel activation by methylxanthines was reported (1) to stimulate the process of excitation coupling in muscle cells, (2) to augment the excitability of neurons and thus their capacity to release neurotransmitters, and also (3) to improve their survival. Here, we address the mechanisms by which methylxanthines control RyR activation and we consider the pharmacological consequences of this activation, in muscle and neuronal cells.


Anlotinib exerts an anti-T-cell acute lymphoblastic leukemia effect in vitro and in vivo.
Despite some progress having been made regarding the treatment of T-cell acute lymphoblastic leukemia (T-ALL), the prognosis of T-ALL, particularly adult T-ALL, is still poor. Identifying novel, effective anti-T-ALL drugs is of great significance. Anlotinib, an oral tyrosine kinase inhibitor currently utilized in the treatment of lung cancer, exhibited a promising anti-T-ALL effect. A comprehensive study should therefore be conducted to explore both the in vitro as well as in vivo mechanisms of the anti-T-ALL effects of anlotinib. CCK8 assays and flow cytometry were employed to investigate the viability, cell cycle distribution, and apoptosis of T-ALL cell lines when treated with anlotinib. T-ALL xenograft mouse models were established to examine the in vivo antileukemic effects of anlotinib. Cellular and molecular analysis of T-ALL were conducted to define the underlying mechanisms. In vitro, anlotinib significantly inhibited the viability, induced G2/M phase arrest and apoptosis in T-ALL cell lines in a concentration-dependent pattern. In vivo, anlotinib also demonstrated a strong anti-tumor effect at doses that are well-tolerated. Interestingly, anlotinib could decrease the protein levels of the intracellular domains of NOTCH1 (ICN1) and c-Myc, two important targets for T-ALL. Mechanistically, anlotinib-induced c-Myc reduction was associated with proteasome-mediated degradation, while the ICN1 reduction was not due to protein degradation or transcriptional repression. The present study showed that anlotinib may be a promising anti-T-ALL candidate drug, and simultaneous reduction of the protein levels of both ICN1 and c-Myc may contribute to the anti-T-ALL efficacy of anlotinib.


Zinc adaptation and resistance to cadmium toxicity in mammalian cells: molecular insight by proteomic analysis.
To identify proteins involved in cellular adaptive responses to zinc, a comparative proteome analysis between a previously developed high zinc- and cadmium-resistant human epithelial cell line (high zinc-resistant HeLa cells, HZR) and the parental HeLa cells has been carried out. Differentially produced proteins included cochaperones, proteins associated with oxido-reductase activities, and ubiquitin. Biochemical pathways to which these proteins belong were probed for their involvement in the resistance of both cell lines against cadmium toxicity. Among ER stressors, thapsigargin sensitized HZR cells, but not HeLa cells, to cadmium toxicity more acutely than tunicamycin, implying that these cells heavily relied on proper intracellular calcium distribution. The similar sensitivity of both HeLa and HZR cells to inhibitors of the proteasome, such as MG-132 or lactacystin, excluded improved proteasome activity as a mechanism associated with zinc adaptation of HZR cells. The enzyme 4-hydroxyphenylpyruvate dioxygenase (HPPD) was overproduced in HZR cells as compared to HeLa cells. It transforms HPP to homogentisate in the second step of tyrosine catabolism. Inhibition of HPPD decreased the resistance of HZR cells against cadmium, but not that of HeLa cells, suggesting that adaptation to zinc overload and increased HPP removal are linked in HZR cells.


Crystallization and preliminary X-ray analysis of GlcNAc alpha 1,4Gal-releasing endo-beta-galactosidase from Clostridium perfringens.
The unique clostridial endo-beta-galactosidase (Endo-beta-Gal(GnGa)) capable of releasing the disaccharide GlcNAc alpha 1,4Gal from O-glycans expressed in the gastric gland mucous cell-type mucin has been crystallized. The crystal belongs to space group P6(3), with unit-cell parameters a = 160.4, c = 86.1 A. Under cryocooled conditions and using a synchrotron X-ray source, the crystals diffract to 1.82 A resolution. The asymmetric unit contains two or three molecules.


Allergy to honeybee body components: distinction from bee venom sensitivity.
Two patients with inhalant allergy to whole bee body components are described, documented by positive skin tests to whole bee body extract and the presence of serum bee body-specific IgE. Both had evidence of mild sensitivity to bee venom. RAST inhibition studies indicated that the IgE antibodies directed at whole bee body components and bee venom were distinct. These observations suggest that venom allergy from insect stings and bee body inhalant allergy are caused by different antigens.


Congenital left atrial aneurysm in an infant.
Congenital left atrial aneurysm, without associated cardiac abnormalities, is a rare defect. We report the case of a large left atrial aneurysm filling almost the entire left hemothorax. This is the first report of this anomaly occurring in an infant less than 1 year of age.


Persistent and De Novo GERD After Sleeve Gastrectomy: Manometric and pH-Impedance Study Findings.
One concerning disadvantage of sleeve gastrectomy (SG) technique as a treatment for obesity is the risk of developing gastroesophageal reflux disease (GERD) or worsening its symptoms after surgery. The purpose of this study is to describe the changes in manometry and 24-h pHmetry with impedance in bariatric patients after SG. Retrospective analysis of 225 patients who underwent SG and were subjected to manometry, 24-h phmetry, and 24-h impedance between 2006 and 2016. Patients were subdivided into groups based on the test performed: 66 patients with manometry, 60 with 24-h pHmetry and 53 with 24-h impedance. Descriptive and analytical statistics were used for analysis. Manometry results showed a decrease in LES pressure from 12.26 ± 6.87 to 8.88 ± 6.28 mmHg in patients after surgery, de novo LES incompetence in 53.33%, worsening of symptoms in 71.4%, increase of inverted gastroesophageal gradient from 6.06% to 48.48% (p<0.05), and a rise in esophageal dysmotility from 18.19% to 37.87%. 24-h pHmetry showed an increase of DeMeester score from 16.71 ± 12.78 to 42.88 ± 32.08 after SG and 79.48% of de novo GERD. Ambulatory 24-h impedance showed an increase of pathological reflux from 47.17% to 88.67% postoperatively. Laparoscopic SG is associated with a higher proportion of abnormal DeMeester score and greater incompetence of the LES in post-SG esophageal tests. Hence, SG may predispose the development of gastroesophageal reflux in the postoperative period.


Fast and New Microbiological Method for Evaluating the Potency of Marbofloxacin-Based Tablets.
Marbofloxacin (MAR) is an antimicrobial belonging to the fluoroquinolone class and is marketed in the form of tablets. There is no microbiological monograph in official compendia to evaluate the potency of the final product. Furthermore, the literature is scarce in this context. The objective is to develop and validate a microbiological method by turbidimetry to evaluate the potency of MAR-based tablets, based on the principles of green analytical chemistry. Purified water-ethanol (9 + 1, v/v) was used as diluent to prepare the MAR solutions at concentrations of 0.25, 0.8, and 2.56 µg/mL, brain heart infusion (BHI) broth as culture media, E. coli American Type Culture Collection (ATCC) 25922 at 10% and incubation in a shaker at 37°C for 4 h were used in the method. The method was: linear in the range of 0.25 to 2.56 µg/mL; selective against the sample adjuvants; precise (intra-day RSD 2.26%, inter-day RSD 3.49%, and inter-analyst RSD 3.59%); accurate with a recovery of 100.20%; and robust against changes in culture medium volume in the tube, shaker rotation, and incubation temperature in the shaker. The potency of MAR tablets was 96.98% using the proposed method. The turbidimetric method developed is a new, fast, and optimized option to the routine QC of MAR in tablets, since it allows the evaluation of the drug's potency in the final product and can be used to complement the results of the physicochemical analysis, in addition to being a green and sustainable alternative. The work shows an ecological and green alternative to the routine microbiological pharmaceutical analysis of MAR tablets.


Patterns of recurrence of rectal cancer after potentially curative surgery.
The results of surgical treatment alone for 142 cases of carcinoma of the rectum and rectosigmoid from the Massachusetts General Hospital were reviewed. The incidence of local failure as any component of failure was found to be strongly dependent on the pathologic stage, and for Dukes' A was 8.0% (3/39); Dukes' B, 31% (18/59), and Dukes' C, 50% (22/44). The incidence of local failure for tumors without lymph node metastasis was 17% with only microscopic extension through the wall (modified Astler-Coller Stage MAC-B2m), but increased to 54% in tumors that were adherent to or invading adjacent organs and structures (MAC-B3). Similarly, in tumors with positive lymph nodes, there was a 36% incidence of local failure for tumors confined to the wall or with only microscopic extension through the wall (MAC-C1/C2m), compared to a 67% incidence for tumors with adherence or involvement of adjacent organs (MAC-C3). Other predictors of local recurrence were the tumor location, grade, number of lymph nodes, and blood vessel invasion. The pathologic factors predicting distant metastasis are also presented. Five-year survival for Dukes' A was 77% (30/39); Dukes' B, 44% (26/59); and Dukes' C, 23% (10/44). The implications for future adjuvant therapy based on the identification of patients with the highest risk for local and distant failure are discussed.


Superior esterolytic activity in environmental Lactococcus lactis strains is linked to the presence of the SGNH hydrolase family of esterases.
Lactococcus lactis strains are widely used in the dairy industry in fermentation processes for production of cheese and fermented milks. However, the esterolytic activity of L. lactis is not generally considered high. For this reason, purified microbial lipases and esterases are often added in certain dairy processes to generate specific flavors in the final food product. This work demonstrates the superior esterolytic activity of a collection of L. lactis strains isolated from different environmental sources compared with that of dairy-derived strains. It provides further evidence of the more diverse metabolic capabilities displayed by L. lactis strains from environmental sources compared to their domesticated dairy counterparts. Furthermore, the presence of a 1,287-bp gene encoding a 428-amino acid SGNH hydrolase in the high-esterolytic environmental strains suggests a possible link between superior esterolytic activity and the presence of the esterase from the SGNH hydrolase family.


Coagulation activation and expression of CD40 ligand on platelets upon in vitro lipopolysaccharide-challenge in patients with unstable angina.
Elevated markers of inflammation and coagulation are found in patients with coronary heart disease. A role of inflammatory stimulation on coagulation time and expression of CD40 ligand on platelets in acute coronary syndromes has not been described yet. Whole blood samples of 9 patients with coronary heart disease and stable angina, 10 patients with unstable angina, 7 patients with acute myocardial infarction and 7 patients without coronary heart disease were incubated with lipopolysaccharide (LPS). Coagulation time was measured in arterial and coronary blood with the ReoRox(R), a viscometric whole blood coagulometer. CD40L and CD62P expression on platelets and platelet-monocyte aggregates were measured by flow cytometry. Without LPS, patients with unstable angina showed a significantly decreased coagulation time in arterial and coronary blood compared to patients without coronary heart disease. After incubation with LPS, in patients with unstable angina, a significantly decreased coagulation time in coronary blood was observed compared to patients with stable angina or patients without coronary heart disease. CD40L expression on platelets in patients with unstable angina was significantly higher in arterial and coronary blood compared to patients with stable angina. No significant differences between the patient groups were observed concerning CD62P expression on platelets, tissue factor binding on monocytes, platelet-monocyte aggregates and plasma levels of platelet factor 4. Patients with unstable angina show an enhanced coagulation activation and an upregulation of CD40L on platelets. This may be of importance in the understanding of coronary plaque rupture and formation of coronary thrombosis.


Potentially avoidable causes of hospitalisation in people with dementia: contemporaneous associations by stage of dementia in a South London clinical cohort.
To estimate the frequency of all-cause and ambulatory care sensitive condition (ACSCs)-related hospitalisations among individuals with dementia. In addition, to investigate differences by stage of dementia based on recorded cognitive function. Data from a large London dementia care clinical case register, linked to a national hospitalisation database. Individuals aged ≥65 years with a confirmed dementia diagnosis with recorded cognitive function. Acute general hospital admissions were evaluated within 6 months of a randomly selected cognitive function score in patients with a clinical diagnosis of dementia. To evaluate associations between ACSC-related hospital admissions (overall and individual ACSCs) and stage of dementia, an ordinal regression was performed, modelling stage of dementia as the dependant variable (to facilitate efficient model selection, with no implication concerning the direction of causality). Of the 5294 people with dementia, 2993 (56.5%) had at least one hospitalisation during a 12-month period of evaluation, and 1192 (22.5%) had an ACSC-related admission. Proportions with an all-cause or ACSC-related hospitalisation were greater in the groups with more advanced dementia (all-cause 53.9%, 57.1% and 60.9%, p 0.002; ACSC-related 19.5%, 24.0% and 25.3%, p<0.0001 in the mild, moderate and severe groups, respectively). An ACSC-related admission was associated with 1.3-fold (95% CI 1.1 to 1.5) increased odds of more severe dementia after adjusting for demographic factors. Concerning admissions for individual ACSCs, the most common ACSC was urinary tract infection /pyelonephritis (9.8% of hospitalised patients) followed by pneumonia (7.1%); in an adjusted model, these were each associated with 1.4-fold increased odds of more severe dementia (95% CI 1.2 to 1.7 and 1.1 to 1.7, respectively). Potentially avoidable hospitalisations were common in people with dementia, particularly in those with greater cognitive impairment. Our results call for greater attention to the extent of cognitive status impairment, and not just dementia diagnosis, when evaluating measures to reduce the risk of potentially avoidable hospitalisations.


Production of monoclonal antibodies to rat lung angiotensin-converting enzyme.
Several monoclonal antibodies to rat lung angiotensin-converting enzyme (ACE) have been produced. The antibodies are of the IgG class, do not inhibit ACE catalytic activity, and do not cross-react with the human or bovine enzyme. They bind in a curvilinear fashion to lung capillary endothelium by immunofluorescence microscopy, and radiolabeled antibody localizes in lung and other organs after intravenous injection into living rats. Each antibody tested appears to bind preferentially to lung rather than kidney ACE by ELISA, a finding supported by weak or absent immunofluorescence of kidney slices in vitro. These antibodies may be used to probe structural differences between ACE in various tissues and, by quantitating changes in accumulation of radiolabeled antibody in experimental models of lung injury, should complement functional measurements in determining the presence of subtle and progressive endothelial damage.


Molecular characterization of an endophytic Phomopsisliquidambaris CBR-15 from Cryptolepis buchanani Roem. and impact of culture media on biosynthesis of antimicrobial metabolites.
An endophytic fungus Phomopsis liquidambaris CBR-15, was isolated from Cryptolepis buchanani Roem. (Asclepiadaceae) and identified by its characteristic culture morphology and molecular analysis of the ITS region of rDNA and intervening 5.8S rRNA gene. The impact of different culture media on biosynthesis of antimicrobial metabolites was tested by disc diffusion assay. Polyketide synthase gene (PKS) of the endophytic fungus was investigated using three pairs of degenerate primers LC1-LC2c, LC3-LC5c and KS3-KS4c by PCR. TLC-bioautography method was employed to detect the antimicrobial metabolites. Antimicrobial metabolites fractionated with ethyl acetate extract showed significant antimicrobial activity against the test bacteria and fungi. Biosynthesis of antimicrobial metabolites was optimum as depicted by zone of inhibition from ethyl acetate extract cultured in potato dextrose broth. Strain CBR-15 was identified as Phomopsisliquidambaris and PKS genes of the fungus were amplified with LC3-LC5c and KS3-KS4c sets of degenerate primers. These findings suggest that endophytic P.liquidambaris CBR-15 harbor iterative type I fungal PKS gene domain which indicates the biosynthetic potential of endophytic fungi as producers of natural antimicrobial metabolites. The study also demonstrates the utilization and optimization of different culture media which best supports for the biosynthesis of the antimicrobial metabolites from P.liquidambaris.


l-Cysteine and glutathione restore the modulation of rat frontal cortex Na+, K+ -ATPase activity induced by aspartame metabolites.
Studies have suggested that aspartame (ASP) ingestion is implicated in neurological problems. The aim of this study was to evaluate rat frontal cortex Na+, K+ -ATPase and Mg2+ -ATPase activities after incubation with ASP or each of its metabolites, phenylalanine (Phe), methanol (MeOH) and aspartic acid (asp) separately. Suckling rat frontal cortex homogenates or pure Na+, K+ -ATPase were incubated with ASP metabolites. Na+, K+ -ATPase and Mg2+ -ATPase activities were measured spectrophotometrically. Incubation of frontal cortex homogenate or pure Na+, K+ -ATPase with various ASP concentrations as expected in the cerebrospinal fluid (CSF) after ASP consumption of 34, 150 or 200mg/kg, decreased the frontal cortex enzyme activity by 33%, 53% or 57%, respectively, whereas pure enzyme was remarkably stimulated. Moreover, incubation of frontal cortex homogenate with each one of the expected ASP metabolites in the CSF, except MeOH, which are related to the intake of the above mentioned doses of the sweetener, resulted in an activation of the membrane Na+, K+ -ATPase, as well as pure enzyme. Frontal cortex Mg2+-ATPase remained unaltered. Addition of l-cysteine (cys) or reduced glutathione (GSH) to ASP metabolites mixtures, corresponding to 150 or 200mg/kg doses of the sweetener, completely or partially restored to normal the modulated membrane and pure Na+, K+ -ATPase activities. CSF concentrations of the sum of ASP metabolites corresponding to the intake of common, abuse or toxic doses (34 or 150 or 200mg/kg, respectively) of the additive significantly increased rat frontal cortex Na+, K+ -ATPase and pure enzyme activities. Cys or GSH completely or partially restored to normal both enzyme activities, possibly due to amelioration of the cellular GSH reduction from the action of MeOH, a metabolite of the sweetener and/or by their scavenging effect.


A Review of Telemedicine Practice Guidelines for COVID-19 and Global Emergencies.
An incorrect version of this manuscript was published online and so has been withdrawn.


Experimental Insight into the Hemodynamics and Perfusion of Radiological Contrast in Patent and Non-patent Aortic Dissection Models.
In a curved vessel such as the aortic arch, the velocity profile closer to the aortic root is normally skewed towards the inner curvature wall, while further downstream along the curve, the velocity profile becomes skewed towards the outer wall. In an aortic dissection (AD) disease, blood velocities in the true lumen (TL) and false lumen (FL) are hypothesized to depend on the proximity of the entry tear to the root of aortic arch. Faster velocity in the FL can lead to higher hemodynamic loading, and pose tearing risk. Furthermore, the luminal velocities control the perfusion rate of radiological contrast media during diagnostic imaging. The objective in this study is to investigate the effect of AD disease morphology and configuration on the blood velocity field in the TL and FL, and on the relative perfusion of radiological enhancement agents through the dissection. Eight in vitro models were studied, including patent and non-patent FL configurations. Particle image velocimetry (PIV) was used to quantify the AD velocity field, while laser-induced fluorescence (LIF) was implemented to visualize dynamical flow phenomena and to quantify the perfusion of injected dye, in mimicry of contrast-enhanced computed tomography (CT). The location of the proximal entry tear along the aortic arch in a patent FL had a dramatic impact on whether the blood velocity was higher in the TL or FL. The luminal velocities were dependent on the entry/reentry tear size combination, with the smaller tear (whether distal or proximal) setting the upper limit on the maximal flow velocity in the FL. Upon merging near the distal reentry tear, the TL/FL velocity differential gave rise to the roll up and shedding of shear layer vortices that convected downstream in close proximity to the wall of the non-dissected aorta. In a non-patent FL, the flow velocity was practically null with all the blood passing through the TL. LIF imaging showed much slower perfusion of contrast dye in the FL compared to the TL. In a patent FL, however, dye had a comparable perfusion rate appearing around the same time as in the TL. Blood velocities in the TL and FL were highly sensitive to the exact dissection configuration. Geometric case A1R, which had its proximal entry tear located further downstream along the aortic arch, and had its entry and reentry tears sufficiently sized, exhibited the highest FL flow velocity among the tested models, and it was also higher than in the TL, which suggest that this configuration had elevated hemodynamic loading and risk for tearing. In contrast-enhanced diagnostic imaging, a time-delayed acquisition protocol is recommended to improve the detection of suspected cases with a non-patent FL.


Proteomic and peptidomic differences and similarities between four muscle types from New Zealand raised Angus steers.
Four muscles from New Zealand-raised Angus steers were evaluated (musculus semitendinosus, m. longissimus thoracis et lumborum, m. psoas major and m. infraspinatus) to test their differences and common features in protein and peptide abundances. The ultimate goal of such a comparison is to match muscle types to products with targeted properties. Protein profiling based on two-dimensional electrophoresis showed that the overall profiles were similar, but, between muscle types, significant (p<0.05) intensity differences were observed in twenty four protein spots. Profiling of endogenous peptides allowed characterisation of 346 peptides. Quantitative analysis showed a clear distinction between the muscle types. Forty-four peptides were identified that showed a statistically significant (p<0.05) and substantial (>2-fold change) difference between at least two muscle types. These analyses demonstrate substantial similarities between these four muscle types, but also clear distinctions in their profiles; specifically a 25% difference between at least two muscles at the peptidomic level, and a 14% difference at the proteomic level.


Effects and side-effects of a brief overcorrection procedure in reducing multiple self-stimulatory behaviour: a single case analysis.
The use of positive practice overcorrection to reduce the classroom self-stimulatory behaviour of a mentally retarded child is described. An initial attempt to reduce four stereotyped hand movements through reinforcement of incompatible behaviour resulted in a decrease in three types of behaviour and an increase in the fourth. When one minute of positive practice overcorrection was then made contingent upon the increased behaviour, it was immediately reduced to near-zero levels with accompanying low occurrences of all other behaviour forms. Response suppression was durable following the evaluation of the overcorrection procedure at 2- and 3.5-month follow-ups.


Probing the effects of N-terminal acetylation on α-synuclein structure, aggregation and cytotoxicity.
Parkinson's disease is associated with the aberrant aggregation of α-synuclein within brain cells. Although the causes of this process are still unclear, post-translational modifications of α-synuclein are likely to play a modulatory role. Since α-synuclein is constitutively N-terminally acetylated, we previously investigated how this protein modification affects the aggregation behavior of the protein using a variety of methods in vitro and in cell systems. This chapter describes the production of N-terminally acetylated (NTA) α-synuclein, the preparation of different seeds of NTA α-synuclein for aggregation assays and the experimental methods for the kinetic analysis of the aggregation process of NTA α-synuclein. We also detail our protocol to evaluate the effects of preformed protofibrils of NTA α-synuclein in cell-based assays. These methods can be applied to study other post-translational modifications of α-synuclein, or adapted for the study of N-acetylation of other aggregation-prone proteins.


Defective stromal cell function in a mouse model of infusion-induced bone marrow failure.
To study bone marrow (BM) stromal damage in a mouse model of infusion-induced BM failure. Sublethally irradiated CByB6F1 mice were infused with 5 x 10(6) C57BL/6 (B6) lymph node (LN) cells. Recipient BM cells were taken at 3, 7, 10, and 14 days following LN infusion and were cultured in vitro in alpha-modified Eagle media for 2-3 weeks. Peripheral blood and was analyzed by complete blood counts while BM lymphocyte infiltration/expansion was analyzed by flow cytometry. Marrow cells from affected and control mice were mixed and cultured in vitro to test nonspecific stromal damage. Donor lymphocytes infiltrated host BM within 3-7 days and expanded significantly between 7 and 10 days, concurrent with the development of leukopenia, thrombocytopenia, and marrow hypoplasia. BM cells from mice at 7, 10, and 14 days after B6-LN cell infusion were progressively defective in forming stromal feeder layers. A 1:1 mixture of BM cells from affected CByB6F1 mice and normal B6 mice failed to form an effective stromal feeder layer that could support cobblestone colony formation, indicating that lymphocytes in the BM of affected CByB6F1 mice were able to damage stromal cells in the normal B6 BM. Activated lymphocytes destroy both hematopoietic and stromal cells as innocent bystanders in the infusion-induced BM failure model.


Roles of the trkB and trkC gene products of Escherichia coli in K+ transport.
Mutations at the trkB and trkC loci of Escherichia coli produce an abnormal efflux of K+. The mutations are partially dominant in diploids and revert frequently by what appears to be intragenic suppression to the null state. The mutations can be reverted by insertion of Tn10 into the mutated gene, and spontaneous revertants are fully recessive to the mutant allele in diploids. K+ efflux produced by NEM* and by DNP* persists in strains with presumed null mutations at either locus, indicating neither gene product is the primary target for the effect of these inhibitors on K+ efflux. The results are consistent with the view that trkB and trkC encode independent systems for K+ efflux. Mutations at these loci alter regulation of the process so that K+ efflux occurs inappropriately. A second mutation to the null state abolishes this abnormal K+ efflux. These genes may encode K+/H+ antiporters, an activity postulated to mediate K+ efflux and demonstrated to exist in E. coli and other bacteria.