The in vitro and in vivo investigation of a novel small chamber dry powder inhalation delivery system for preclinical dosing to rats.
This research describes a novel "minitower" dry powder delivery system for nose-only delivery of dry powder aerosols to spontaneously breathing rats. The minitower system forces pressurized air through pre-filled capsules to deliver aerosolized drug to four nose ports; three of which house spontaneously breathing rats, with the fourth used as a control. Within each port are vent filters which capture drug that was not inhaled for further quantitation. These vent filters along with a novel control system referred to as the "artificial rat lung", allow for the theoretical amount of drug delivered and subsequently inhaled by each rat to be calculated. In vitro and in vivo studies have demonstrated this system's ability to deliver aerosolized drug to rats. The in vitro study showed that ∼30% of the starting dose reached the 4 ports and was available for inhalation. During in-vivo studies, rats inhaled ∼34% of the delivered dose. Of the estimated inhaled dose, 12-18% was detectable in the various tissue samples, with over 30% of the recovered dose found in the rat's lungs. Results show that this system is capable of reproducibly delivering drug to the lungs of spontaneously breathing rats. Advantages over current delivery methods include being amenable to the administration of multiple doses and using less (milligram) amount of starting material. In addition, this technique avoids anesthesia which is typically required for instillation or insufflation, and thus has the potential as an efficient and noninvasive aerosol delivery method for preclinical drug development.


A retrospective serologic survey for Anaplasma spp. infection in three bighorn sheep (Ovis canadensis) populations in California.
Using an indirect immunofluorescence assay, we determined the prevalence of Anaplasma-reactive antibody in three herds of bighorn sheep, each a different subspecies and occupying a different habitat in California (USA). Antibodies to Anaplasma spp. were identified in none of twenty California bighorn (Ovis canadensis californiana) sampled from the Mt. Baxter herd, 11 of 17 peninsular bighorn (O. canadensis cremnobates) sampled in the Santa Rosa Mountains, and all 20 desert bighorn (O. canadensis nelsoni) sampled at Old Dad Peak/Kelso Mountains. Based on an assay and an adsorption technique, the titers most likely were due to Anaplasma ovis. The presence and species of tick vectors in each of the habitats, and the presence or absence of deer or livestock were identified as factors potentially influencing seroprevalence of antibodies.


Does the mode of questionnaire administration affect the reporting of urinary symptoms?
To assess the effect of modes of administration (self-administered questionnaires, oral face-to-face interview, and telephone interview) on responses to the American Urological Association Symptom Index (AUASI) in randomly selected community men. An age-stratified random sample of 475 white male residents of Olmsted County, Minnesota, aged 40 to 79 years, without prior prostate surgery or prostate cancer were queried about urinary symptom frequency twice at baseline and twice approximately 2 years later using questions with wording similar to the AUASI: At baseline and first follow-up, questionnaires were self-administered initially, followed by a structured interview by a female urology nurse within 2 to 28 weeks. A subset of 200 randomly selected men received a telephone interview by a female research assistant following the self-administered questionnaire given at a second follow-up approximately 4 years after baseline. Mean symptom scores obtained by oral interview were 1 to 2 points lower than those from self-administered questionnaires (P < 0.01). In a random subset (n = 200) interviewed by telephone, mean AUASI scores were as much as 4 points lower than those from self-completed questionnaires. Values of the AUASI obtained by interviewer administration may be lower than those obtained by self-administered questionnaires. When assessment of change in urinary symptoms over time is of interest, the same standardized method of questionnaire administration should be used at baseline and follow-up evaluations to avoid introducing artifactual differences related to the mode of administration.


The Neurotoxic Effect of β-Amyloid Is Accompanied by Changes in the Mitochondrial Dynamics and Autophagy in Neurons and Brain Endothelial Cells in the Experimental Model of Alzheimer's Disease.
A comparative assessment of the expression of the mitochondrial fission marker Drp1 and the autophagy marker LC3 in neurons and endothelial cells in the hippocampus and entorhinal cortex during progression of cognitive deficit was performed in animals with intrahippocampal administration of β-amyloid. In both brain regions, the expression of Drp1 and LC3 in neuronal and endothelial cells was enhanced. The peak of cognitive impairment corresponded to the maximum expression of Drp1 and LC3 in hippocampal neurons and was preceded by an increase in the number of Drp1+ and LC3+ endothelial cells in this brain region. These data attests to a possible role of aberrant mitochondrial dynamics and autophagy of endothelial cells in the impairment of brain plasticity in the Alzheimer's type neurodegeneration.


Automatic data extraction from 24 hour blood pressure measurement reports of a large multicenter clinical trial.
Ambulatory blood pressure monitoring (ABPM) is usually reported in descriptive values such as circadian averages and standard deviations. Making use of the original, individual blood pressure measurements may be advantageous, particularly for research purposes, as this increases the flexibility of the analytical process, enables alternative statistical analyses and provide novel insights. Here we describe the development of a new multistep, hierarchical data extraction algorithm to collect raw data from .pdf reports and text files as part of a large multi-center clinical study. Original reports were saved in a nested file system, from which they were automatically extracted, read and saved into databases with custom made programs written in Python 3. Data were further processed, cleaned and relevant descriptive statistics such as averages and standard deviations calculated according to a variety of definitions of day- and night-time. Additionally, data control mechanisms for manual review of the data and programmatic auto-detection of extraction errors was implemented as part of the project. The developed algorithm extracted 97% of the data automatically, the missing data consisted mostly of reports that were saved incorrectly or not formatted in the specified way. Manual checks comparing samples of the extracted data to original reports indicated a high level of accuracy of the extracted data, no errors introduced due to flaws in the extraction software were detected in the extracted dataset. The developed multistep, hierarchical data extraction algorithm facilitated collection from different file formats and paired with database cleaning and data processing steps led to an effective and accurate assembly of raw ABPM data for further and adjustable analyses. Manual work was minimized while data quality was ensured with standardized, reproducible procedures.


Radical prostatectomy: long-term cancer control and recovery of sexual and urinary function ("trifecta").
Radical prostatectomy (RP) disrupts the natural history of prostate cancer. However, it could be a significant source of long-term incontinence and potency morbidity. We studied the long-term cancer survival results and the probabilities of achieving ideal "trifecta" outcomes (cancer control, continence, and potency) after this surgical procedure. A total of 1746 intervention-naive patients with clinically localized newly diagnosed prostate cancer underwent RP with curative intent beginning in 1983. The mean follow-up time was 6 years (interquartile range, 3-9). The successive probabilities of achieving ideal trifecta outcomes for up to 24 months following RP versus experiencing biochemical recurrence were estimated using the cumulative incidence method. Additionally, long-term cancer control was assessed by preoperative and postoperative factors. Surgical excision controlled prostate cancer effectively in 1441 (83%) of the 1746 patients studied. At 5, 10, and 15 years, respectively, 82%, 77%, and 75% of patients were free from disease progression. Cancer-specific survival was 99%, 95% and 89%, respectively, at 5, 10, and 15 years. In men with disease progression, the 15-year probabilities of death from prostate cancer versus other causes were similar (32% and 33%, respectively). At 24 months, 60% of patients were potent, continent, and free of cancer, and 12% had experienced recurrence. Use of RP provided excellent long-term cancer control. At 15 years, only 11% of patients had died of prostate cancer. Cancer control was good even for patients with adverse prognostic features. The probability of death from cancer was similar to other causes after disease progression. By 2 years, 60% of men were continent, potent, and cancer free.


Efficient evaluation of photodynamic therapy on tumor based on deep learning.
Photodynamic therapy (PDT) is a non-invasive treatment method for treating tumors. Under laser irradiation, photosensitizers in tumor tissues generate biotoxic reactive oxygen, which can kill tumor cells. The traditional live/dead staining method of evaluating the cell mortality caused by PDT mainly depends on manual counting, which is time-consuming and relies on dye quality. In this paper, we have constructed a dataset of cells after PDT treatment and trained the cell detection model YOLOv3 to count both the dead and live cells. YOLO is a real time AI object detection algorithm. The achieved results demonstrate that the proposed method has a good performance in cell detection, with a mean average precision (mAP) of 94% for live cells and 71.3% for dead cells. This approach can efficiently evaluate the effectiveness of PDT treatment, thus speeding up treatment development effectively.


Purification of GFP fusion proteins with high purity and yield by monoclonal antibody-coupled affinity column chromatography.
GFP has often been used as a marker of gene expression, protein localization in living and fixed tissues as well as for protein targeting in intact cells and organisms. Monitoring foreign protein expression via GFP fusion is also very appealing for bioprocess applications. Many cells, including bacterial, fungal, plant, insect and mammalian cells, can express recombinant GFP (rGFP) efficiently. Several methods and procedures have been developed to purify the rGFP or recombinant proteins fused with GFP tag. However, most current GFP purification methods are limited by poor yields and low purity. In the current study, we developed an improved purification method, utilizing a FMU-GFP.5 monoclonal antibody (mAb) to GFP together with a mAb-coupled affinity chromatography column. The method resulted in a sample that was highly pure (more than 97% homogeneity) and had a sample yield of about 90%. Moreover, the GFP epitope permitted the isolation of almost all the active recombinant target proteins fused with GFP, directly and easily, from the crude cellular sources. Our data suggests this method is more efficient than any currently available method for purification of GFP protein.


Integrated tool for risk assessment in agricultural management of soil erosion and losses of phosphorus and nitrogen.
In recent years, increased attention has been focused on models for risk assessment of source areas in agricultural landscapes. Among the simplest of such models are index tools, which have been developed particularly for phosphorus (P) and to some extent nitrogen (N). However, only a few studies have considered the development of an integrated management strategy that includes erosion and losses of both P and N. Accordingly, the major objective of this study was to initiate the development of an integrated risk assessment tool, consisting of indices for erosion, P and N. The strategy used to create the integrated tool was based on the assumption that all input data at field scale should be readily available either from ordinary agricultural statistics or from the farmer. The results from using the indices in a pilot case study catchment illustrated that losses of P and N had often different critical source areas. The P index was highest for fields with manure application and/or high soil P status or with autumn ploughing, and the N index was highest for fields with excessive N application. The integrated risk was greatest for areas with manure application and some areas with a high erosion risk in combination with high nutrient application rate. Additionally, four different management options were assessed: (1) reduced fertilisation, (2) catch crops, (3) autumn ploughing, and (4) no autumn ploughing. The results verified that reduced nutrient application and stubble during autumn and winter led to the largest decrease in index values, and it was also apparent that management changes in high-risk areas had the greatest impact on the indices. Overall, our findings indicate that the present integrated risk assessment tool with readily available input data can be used to rank farm fields according to risk of soil erosion and losses of P and N.


An enzyme-linked immunoassay for thyroid microsomal antibodies.
An enzyme-linked immunoassay (ELISA) for microsomal antibody is described. The method was found to be rapid, sensitive and precise and analysis of 115 serum samples showed good correlation between the ELISA and the conventional tanned red cell haemagglutination test. The presence of thyroglobulin antibody, rheumatoid factor, antinuclear factor or gastric parietal cell antibodies did not interfere in the microsomal antibody ELISA but some sera with mitochondrial antibody activity appeared to cause a non-specific effect. The ELISA was particularly useful for analysing microsomal antibody production by Hashimoto lymphocyte cultures and in some cases antibody synthesis could be studied in the absence of mitogen. The high capacity of the ELISA combined with its sensitivity suggest that it will be a valuable technique for studying microsomal autoantibody activity both in serum and in lymphocyte cultures.


Evidence for the Maillard reaction in rat lung collagen and its relationship with solubility and age.
This study investigated age-related changes in collagen solubility and collagen-linked fluorescence, and their relationship with the Maillard reaction. As a result of the collagen purification of rat lung samples, we obtained two pools of collagen with different degrees of solubility. The relative distribution of collagen between these two fractions was time-dependent, and the proportion of the smaller and less soluble fraction increased with time (r = 0.73, P < 0.0001). In this fraction, the intensity of fluorescence at Exc 335 nm/Em 385 nm, and the total amount of pentosidine increased with age (r = 0.66, P < 0.002, and r = 0.69, P < 0.01, respectively). The mean values for fluorescence and pentosidine per milligram of collagen were, respectively, six and ten times greater in the less soluble fraction. In this fraction the pentosidine per milligram of collagen increased with age (r = 0.59, P < 0.03). Our results demonstrated the presence of pentosidine in rat lung collagen. Moreover, its accumulation in the less soluble fraction suggested a relationship between Maillard reaction products, physico-chemical changes in collagen solubility, and the ageing process in rat lungs.


Rop, the Sec1/Munc18 homolog in Drosophila, is required for furrow ingression and stable cell shape during cytokinesis.
Physically separating daughter cells during cytokinesis requires contraction of an actin-myosin ring and vesicle-mediated membrane addition at the cleavage furrow. To identify vesicle trafficking proteins that function in cytokinesis, we screened deficiencies and mutations of candidate genes by live imaging the mitotic domains of the Drosophila embryo. In embryos homozygous for some of these deficiencies, we observed several cytokinesis phenotypes, including slow furrow ingression and increased membrane blebbing. We also found that cytokinesis required the Sec1/Munc18 homolog Rop, which interacts with syntaxin and mediates exocytosis at the plasma membrane. In a temperature-sensitive Rop mutant (Rop(TS)), the contractile ring disassembled during furrow ingression, indicating that maintenance of the ring required vesicle addition. Furthermore, in some dividing Rop(TS) cells, the shape of the daughter cells became unstable, causing cytokinesis failure. These results further highlight the importance of vesicle trafficking in animal cytokinesis and show that vesicle fusion influences cell shape during cytokinesis.


Platelet Volume Is Reduced In Metastasing Breast Cancer: Blood Profiles Reveal Significant Shifts.
Platelets play a crucial role in breast cancer (BC) progression and metastases. Mean platelet volume (MPV) is an indicator of platelet activation. The aim of the present study was to assess whether there is a difference in MPV between patients with metastatic BC with liver metastases and those with BC without liver metastases. Between January 2014 and December 2017, 211 metastatic BC patients with synchronous liver metastases and 215 BC patients without metastases were retrospectively analyzed. Patients' clinicopathological characteristics data were collected. MPV levels were reduced in patients with liver metastases compared with those in patients without liver metastases. There were significant differences in MPV levels according to liver metastases status both in premenopausal and in postmenopausal non-TNBC or non-HER2+ patients. Moreover, in postmenopausal HER2+ or TNBC patients, MPV levels were lower in patients with liver metastases compared with those in patients without liver metastases. In the group with non-liver metastasis, platelet distribution width was significantly associated with tumor N stage. In addition, the prevalence of BC liver metastases decreased as MPV quartiles increased. After adjusting for other risk factors, the odds ratios for liver metastases according to MPV quartiles were 1.000, 0.267 (0.134-0.530), 0.072 (0.034-0.152), and 0.137 (0.066-0.281), respectively. MPV is reduced in BC patients with liver metastases compared with that in BC patients without metastases. Moreover, MPV is independently associated with the presence of liver metastases.


Caring for the frail older patient: the doctor-patient-family caregiver relationship.
Much has been written about the complexities of the doctor-patient relationship and the myriad ways that both parties communicate. When patients become more frail, this relationship must expand to include their family caregivers. This addition of a third person complicates the situation in more than an additive fashion. It requires more time and energy on the part of physicians that are not without costs. Balancing both patient and caregiver needs in a time-driven and increasingly competitive world is not easy. Physicians not only need better preparation to meet the challenges of caring for frail older patients, but they also need changes in reimbursement policies so that they can afford to spend the time needed to manage the complexities inherent in the doctor-patient-family caregiver relationship. Although there is uncertainty about timing of these changes, there is no uncertainty about the complexity of the doctor-patient-family caregiver relationship. To make its management easier, the following rules are worth remembering: 1. Carefully assess problems in all domains of patient function and develop plans for managing and monitoring these problems. 2. Recognize that the effects of patient disabilities cycle between patient and caregiver. 3. Pay attention to the needs and concerns of patients and of their caregivers. 4. Provide caregivers with as much information as possible and educate them about "when to worry." 5. Assure caregivers that you are available to answer questions and help them with handling problems that arise in the care of their elders.


Increased incidence of nonfatal cardiovascular events in stroke patients with sleep apnoea: effect of CPAP treatment.
Obstructive sleep apnoea (OSA) is a risk factor for stroke, but little is known about the effect of OSA and continuous positive airway pressure (CPAP) on the incidence of long-term, nonfatal cardiovascular events (CVE) in stroke patients. A prospective observational study was made in 223 patients consecutively admitted for stroke. A sleep study was performed on 166 of them. 31 had an apnoea/hypopnoea index (AHI) <10 events · h(-1); 39 had an AHI between 10 and 19 events · h(-1) and 96 had an AHI ≥ 20 events · h(-1). CPAP treatment was offered when AHI was ≥ 20 events · h(-1). Patients were followed up for 7 yrs and incident CVE data were recorded. The mean ± SD age of the subjects was 73.3 ± 11 yrs; mean AHI was 26 ± 16.7 events · h(-1). Patients with moderate-to-severe OSA who could not tolerate CPAP (AHI ≥ 20 events · h(-1); n = 68) showed an increased adjusted incidence of nonfatal CVE, especially new ischaemic strokes (hazard ratio 2.87, 95% CI 1.11-7.71; p = 0.03), compared with patients with moderate-to-severe OSA who tolerated CPAP (n = 28), patients with mild disease (AHI 10-19 events · h(-1); n = 36) and patients without OSA (AHI <10 events · h(-1); n = 31). Our results suggest that the presence of moderate-to-severe OSA is associated with an increased long-term incidence of nonfatal CVE in stroke patients and that CPAP reduces the excess of incidence seen in these patients.


Augmenting NMDA receptor signaling boosts experience-dependent neuroplasticity in the adult human brain.
Experience-dependent plasticity is a fundamental property of the brain. It is critical for everyday function, is impaired in a range of neurological and psychiatric disorders, and frequently depends on long-term potentiation (LTP). Preclinical studies suggest that augmenting N-methyl-d-aspartate receptor (NMDAR) signaling may promote experience-dependent plasticity; however, a lack of noninvasive methods has limited our ability to test this idea in humans until recently. We examined the effects of enhancing NMDAR signaling using d-cycloserine (DCS) on a recently developed LTP EEG paradigm that uses high-frequency visual stimulation (HFvS) to induce neural potentiation in visual cortex neurons, as well as on three cognitive tasks: a weather prediction task (WPT), an information integration task (IIT), and a n-back task. The WPT and IIT are learning tasks that require practice with feedback to reach optimal performance. The n-back assesses working memory. Healthy adults were randomized to receive DCS (100 mg; n = 32) or placebo (n = 33); groups were similar in IQ and demographic characteristics. Participants who received DCS showed enhanced potentiation of neural responses following repetitive HFvS, as well as enhanced performance on the WPT and IIT. Groups did not differ on the n-back. Augmenting NMDAR signaling using DCS therefore enhanced activity-dependent plasticity in human adults, as demonstrated by lasting enhancement of neural potentiation following repetitive HFvS and accelerated acquisition of two learning tasks. Results highlight the utility of considering cellular mechanisms underlying distinct cognitive functions when investigating potential cognitive enhancers.


[The Alliance for the Prevention of Colorectal Cancer in Spain. A civil commitment to society].
Colorectal cancer (CRC) is the most common malignant tumor in Spain, when men and women are considered together, and the second leading cause of cancer death. Every week in Spain over 500 cases of CRC are diagnosed, and nearly 260 people die from the disease. Epidemiologic estimations for the coming years show a significant increase in the number of annual cases. CRC is a perfectly preventable tumor and can be cured in 90% of cases if detected in the early stages. Population-based screening programs have been shown to reduce the incidence of CRC and mortality from the disease. Unless early detection programs are established in Spain, it is estimated that in the coming years, 1 out of 20 men and 1 out of 30 women will develop CRC before the age of 75. The Alliance for the Prevention of Colorectal Cancer in Spain is an independent and non-profit organization created in 2008 that integrates patients' associations, altruistic non-governmental organizations and scientific societies. Its main objective is to raise awareness and disseminate information on the social and healthcare importance of CRC in Spain and to promote screening measures, early detection and prevention programs. Health professionals, scientific societies, healthcare institutions and civil society should be sensitized to this highly important health problem that requires the participation of all sectors of society. The early detection of CRC is an issue that affects the whole of society and therefore it is imperative for all sectors to work together.


Label-free detection of cellular drug responses by high-throughput bright-field imaging and machine learning.
In the last decade, high-content screening based on multivariate single-cell imaging has been proven effective in drug discovery to evaluate drug-induced phenotypic variations. Unfortunately, this method inherently requires fluorescent labeling which has several drawbacks. Here we present a label-free method for evaluating cellular drug responses only by high-throughput bright-field imaging with the aid of machine learning algorithms. Specifically, we performed high-throughput bright-field imaging of numerous drug-treated and -untreated cells (N = ~240,000) by optofluidic time-stretch microscopy with high throughput up to 10,000 cells/s and applied machine learning to the cell images to identify their morphological variations which are too subtle for human eyes to detect. Consequently, we achieved a high accuracy of 92% in distinguishing drug-treated and -untreated cells without the need for labeling. Furthermore, we also demonstrated that dose-dependent, drug-induced morphological change from different experiments can be inferred from the classification accuracy of a single classification model. Our work lays the groundwork for label-free drug screening in pharmaceutical science and industry.


CD 152 gene polymorphisms and risk of osteosarcoma in Chinese population.
Osteosarcoma has become a health threat for adolescents and young adults. To identify the genetic risk factor for the malignancy is in urgent need. Several studies have investigated the role of CD 152 polymorphisms in osteosarcoma in a sample of Chinese population. However, the association is poorly defined due to lack of a sufficiently large sample. In this study, we performed a meta-analysis of all CD 152 polymorphisms that had been implicated in osteosarcoma to examine the association. We searched the electronic MEDLINE database until December 31, 2013, to identify the studies regarding the association between CD 152 polymorphisms and osteosarcoma. Inclusion criteria were followed in the selection of eligible study. The genotypic and allelic data were collected from all studies included to evaluate the risk of osteosarcoma (odds ratio, OR). We found statistically significant evidence of the studied CD 152 polymorphisms and increased risk of osteosarcoma in homozygous (OR = 1.79, 95 % CI = 1.40-2.29, P = 0.958), recessive (OR = 1.77, 95 % CI = 1.40-2.25, P = 0.899), and allele model (OR = 1.21, 95 % CI = 1.09-1.34, P = 1.000). This increased risk was also revealed in single nucleotide polymorphism (SNP) +49G>A and SNP 326G>A. Our meta-analysis indicates that there may be an association between CD 152 polymorphisms and risk of osteosarcoma in Chinese population. Further validation of the observation is necessary.


[Immunosuppression and its use in kidney transplantation].
Current immunosuppressive protocols effectively prevent acute rejection of renal allografts. Extensive drug toxicity and the deleterious effects of long-term immunosuppression are associated with significant morbidity and mortality. The purpose of this article is to provide an overview over modern immunosuppressants and their unwanted side effects and to discuss strategies for improved long-term transplant survival. Review of the current topic-related literature and discussion of our own experience. The use of antibody induction together with an initial combination therapy of calcineurin inhibitors, mycophenolate and steroids is recommended and results in excellent early outcomes. Detrimental effects include an increased incidence of infections, malignomas, and cardiovascular diseases. Long-term transplant survival is impaired by extensive drug toxicity and the frequent development of donor specific antibodies. Reduction of overall cumulative exposure to immunosuppressants or the reduction of specific toxic drugs such as calcineurin inhibitors and steroids may improve long-term results. Alternative immunosuppressants like mTOR inhibitors and belatacept appear to be effective and safe but their long-term effects on patient and allograft survival needs to be established in clinical trials. Current immunosuppressants provide effective protection from renal allograft rejection. However, their use is complicated by serious side effects. In the future, development of novel immunosuppressants and optimization of minimization strategies may help to improve long-term success after kidney transplantation.


Photocatalytic Charge-Transfer Complex Enables Hydroarylation of Alkenes for Heterocycle Synthesis.
Here, we report a photocatalytic charge-transfer complex (CTC) strategy for one electron reduction of alkenes using thiolate as a catalytic electron donor. This catalytic CTC system could engage hydroarylation of both activated and unactivated alkenes for the synthesis of various heterocycles. The reactions do not require any photocatalysts or acids and are easy to perform. Mechanistic studies revealed the formation of a CTC between catalytic thiolate and alkene.


Bacterial growth inhibition produced by root canal sealer cements with a calcium hydroxide base.
Inhibition of growth of six bacterial strains produced by two root canal sealers with a calcium hydroxide base, CRCS and Sealapex sealers, is studied. The results are compared with those obtained with two zinc oxide eugenol sealers and one epoxy resin. The inhibition produced with the calcium hydroxide sealers is similar to that obtained with the other sealers. The component of paraformaldehyde in a sealer increases the inhibition significantly.


Hide and run. Arginine-based endoplasmic-reticulum-sorting motifs in the assembly of heteromultimeric membrane proteins.
Arginine-based endoplasmic reticulum (ER)-localization signals are sorting motifs that are involved in the biosynthetic transport of multimeric membrane proteins. After their discovery in the invariant chain of the major histocompatibility complex class II, several hallmarks of these signals have emerged. They occur in polytopic membrane proteins that are subunits of membrane protein complexes; the presence of the signal maintains improperly assembled subunits in the ER by retention or retrieval until it is masked as a result of heteromultimeric assembly. A distinct consensus sequence and their position independence with respect to the distal termini of the protein distinguish them from other ER-sorting motifs. Recognition by the coatomer (COPI) vesicle coat explains ER retrieval. Often, di-leucine endocytic signals occur close to arginine-based signals. Recruitment of 14-3-3 family or PDZ-domain proteins can counteract ER-localization activity, as can phosphorylation. This, and the occurrence of arginine-based signals in alternatively spliced regions, implicates them in the regulated surface expression of multimeric membrane proteins in addition to their function in quality control.


Proliferation capacity of T-lymphocytes is affected transiently after a long-term weight gain in Beagle dogs.
Across species obesity is associated with several disorders but in companion animals little information is available on the impact of chronic obesity on immune competence. The aim of the present study was to investigate whether weight gain and stable obese bodyweight affects the immune cell response. Obesity was induced in eight adult healthy beagle dogs (weight gain group; WGG) by a weight gain period (WGP) of 47 weeks, which was immediately followed by a period (stable period: SP) of stable obesity of 26 weeks. Eight adult healthy beagle dogs were included as a control group (CG) and remained at their ideal bodyweight throughout the entire study. Body composition was measured at five intervening time-points. Concentration of serum leptin and inflammatory cytokines, functionality of lymphocytes and phagocytic activity of neutrophils and monocytes were evaluated at ten intervening time-points. Serum leptin concentration was rising during the WGP in the WGG but went to lower concentrations during the SP. At the end of long-term weight gain, a decreased mitogen-induced proliferation of T-lymphocytes was noted but this alteration seemed to be transient after stabilization of bodyweight. This finding may imply an altered immune response for dogs with different energy balances. However, no systemic low grade inflammation or alteration in other immune cell functions was observed. Consequently it is suggested that the change in energy balance during the onset of obesity (becoming obese versus being obese), evokes an additional obesity-related disorder in dogs, i.e. impaired T-lymphocyte immune function.


Serotonin acts as an up-regulator of acyl-coenzyme A:cholesterol acyltransferase-1 in human monocyte-macrophages.
Acyl-coenzyme A:cholesterol acyltransferase-1 (ACAT-1) converts intracellular free cholesterol into cholesterol ester for storage in lipid droplets and plays an important role in the formation of macrophage-derived foam cells in atherosclerotic lesions. Serotonin (5-HT), a potent vasoconstrictor that is released from activated platelets, increases uptake of oxidized low-density lipoprotein (LDL) by macrophages, leading to foam cell formation, and contributes to the development of atherosclerotic plaque. However, it is not yet known whether 5-HT affects ACAT-1 expression in human monocyte-macrophages as the molecular mechanism of enhanced foam cell formation by 5-HT remains unclear. We examined the effects of 5-HT on ACAT-1 expression during differentiation of cultured human monocytes into macrophages. Expression of ACAT-1 protein but not 5-HT2A receptor increased in a time-dependent manner. 5-HT increased ACAT activity in a concentration-dependent manner after 7 days in primary monocyte culture. Immunoblotting analysis showed that 5-HT at 10 microM increased ACAT-1 protein expression level by two-fold, and this effect was abolished completely by a 5-HT2A receptor antagonist (sarpogrelate), its major metabolite (M-1), a G protein inactivator (GDP-beta-S), a protein kinase C (PKC) inhibitor (rottlerin), a Src family inhibitor (PP2), or a mitogen-activated protein kinase (MAPK) kinase inhibitor (PD98059). Northern blotting analysis indicated that among the four ACAT-1 mRNA transcripts (2.8-, 3.6-, 4.3-, and 7.0-kb), the levels of the 2.8- and 3.6-kb transcripts were selectively up-regulated by approximately 1.7-fold by 5-HT (10 microM). The results of the present study suggested that 5-HT may play a crucial role in macrophage-derived foam cell formation by up-regulating ACAT-1 expression via the 5-HT2A receptor/G protein/c-Src/PKC/MAPK pathway, contributing to the progression of atherosclerotic plaque.


5-HT1C receptors in the serotonergic control of periaqueductal gray induced aversion in rats.
The functional role of brain 5-HT and 5-HT receptor subtypes in periaqueductal gray (PAG) induced aversion has been investigated in rats. Antiaversive effects were found with the serotonin agonists TFMPP, mCPP and DOI but not with RU 24969 which was found to facilitate PAG aversion. The first three serotonin agonists share potent 5-HT1C activity while RU 24969 differs with a high 5-HT1A activity. Proaversive effects were found with the mixed 5-HT1C/5-HT2 antagonists cyproheptadine and ritanserin; this effect was already reported for the mixed 5-HT1C/5-HT2 antagonists metergoline and mianserin and is opposite to the effects of the selective 5-HT2 antagonists ketanserin, pirenperone, trazodone and spiperone. The antiaversive effects of mCPP (1 mg/kg) could be prevented by pretreatment of the animals with mianserin (1 and 10 mg/kg). These results suggest that 5-HT1C receptors play an important role in the serotonergic control of PAG aversion. 5-HT1C receptor activation seems to mediate antiaversive effects whereas acute 5-HT1C receptor blockade appears to facilitate PAG aversion. Functional interactions take place between several receptor types in the in vivo control of PAG aversion, where 5-HT1C receptors appear to play a predominant function.


Epidemiological features of an outbreak of gastroenteritis/cholera in Katsina, Northern Nigeria.
In April 1982, Katsina, in Northern Nigeria, was affected by an outbreak of gastroenteritis associated with Vibrio cholerae serotype 'Ogawa' and 662 patients were admitted to the Katsina General Hospital during a 16-week period. The outbreak affected all ages and both sexes and all parts of the town and its immediate surroundings except the Government Residential Area (GRA). The overall case fatality rate was 7.7%. Male specific case fatality and female specific case fatality rates were 9.7 and 6.2% respectively. 'Adults' and those in the 11-20 and 21-30 age groups accounted for most of the cases. The epidemic curve was of a propagated and protracted nature. About 51.7% of all the patients spent between 2 and 5 days in the hospital. A similar pattern was observed for all age groups regardless of sex. Cholera vibrio 'Ogawa', Shigella spp., Salmonella spp., Proteus spp. and Escherichia coli were isolated from 16 patients. All well-water samples obtained from the compounds of the cases were contaminated with MPN/100 ml index ranging from 540 to greater than 2400. All samples were positive for faecal coliforms. Salmonella spp., Shigella spp., Proteus spp. and non-O, group 1 (non-O 1) V. cholerae were isolated. Water sellers probably facilitated the spread of the outbreak.


Stable long-term recordings from cat peripheral nerves.
A procedure has been developed for the stable long-term recording of nerve signals in unanesthetized mammals, which should have wide application in basic research on the nervous system and also in clinical areas such as the derivation of control signals for powered prostheses. Methods are fully described for constructing devices consisting of (1) Silastic nerve cuffs containing three or more electrodes, (2) coiled leads insulated with Silastic which extend from the cuffs to an integrated circuit socket, (3) a vitreous carbon transcutaneous connector which surrounds the integrated circuit socket and makes a good interface with the skin. Neural activity has been recorded from mammalian nerves for many months during normal behaviour. The peak-to-peak amplitude and latency of the recorded compound action potentials remain stable and may continue at a constant level more or less indefinitely. A tripolar recording configuration between a central lead and the two end leads, which are connected together, permits good rejection of EMG signals from surrounding muscles. The amplitude of single unit potentials increases as the square of the conduction velocity of the nerve fibre. Thus, the largest nerve fibres will dominate the signals recorded during behaviour. The reasons for premature termination of a few experiments are given together with methods for overcoming these problems. For example, platinum-iridium electrodes remain relatively stable, whereas silver wires tend to fracture after being in an animal for several months. This and other relationships are discussed which permit an optimal design of nerve cuffs for a given recording situation.


Promiscuous esterase activities of the C-C hydrolases from Dyella ginsengisoli.
A C-C hydrolase gene (bphD(LA-4)) from strain Dyella ginsengisoli LA-4 was cloned and expressed in Escherichia coli BL21 (DE3). BphD(LA-4) together with another hydrolase MfphA(LA-4), which derived from the same strain, possessed esterase activities. p-Nitrophenyl butyrate was the best substrate for both enzymes. BphD(LA-4) had high catalytic efficiency to p-nitrophenyl benzoate, whereas MfphA(LA-4) had no activity. Homology modeling and docking studies demonstrated that the proper hydrogen bond interaction was important for the reactivity of specific substrate.


Determination of hypoxia and dietary copper mediated sub-lethal toxicity in carp, Cyprinus carpio, at different levels of biological organisation.
Hypoxic events frequently occur in the aquatic environment in association with micro pollutants, including heavy metals. Only a few studies are however available on the uptake and biological responses of heavy metals under hypoxic conditions. To elucidate the phenomenon, mirror carp Cyprinus carpio L. (16.13-16.22 g) were exposed chronically to dietary copper (Cu; 250 and 500 mg kg dry wt.(-1)) for 30 d under normoxic (8.25 mg O(2) L(-1)) and hypoxic (~3 mg O(2) L(-1)) conditions and adopting an integrated approach, sub-lethal biomarker responses were determined at different levels of biological organisation. Level of oxidative DNA damage (as determined by modified Comet assay) showed strong significant difference following exposure to dietary Cu level under normoxic (1.6-fold) as well as under hypoxic condition at both Cu levels (2.1 and 2.5-folds respectively). Significant difference was also observed for haematological parameters (i.e. increased red and white blood cells, haematocrit value and haemoglobin concentration). Quantitative histology revealed alterations in tissues (i.e. liver and gills) for hypoxic and all dietary Cu treatment groups under both normoxic and hypoxic conditions suggesting a compensatory response to these organs (p<0.05). The order of Cu accumulation in tissues (as determined by ICP-OES) was liver>intestine>kidney>gill. Interestingly, SGR under both normoxic and hypoxic conditions reduced with elevating Cu levels (p=0.019). Overall, the results provide evidence for enhanced toxicological responses in fish following exposure to Cu either alone or in combination with hypoxic condition and lends support to the evolving viewpoint that many water quality guidelines should be revisited in terms of new ecotoxicological criteria.


Gamete and embryo donation for research: what might shape the willingness to donate among gamete donors and recipients?
Research using gametes and embryos donated by reproductive and third-party donors contributed to substantial, albeit contentious achievements. The views of gamete donors and recipients on donation for research and the underpinning role of attitudes towards research have been seldom explored and are yet to be incorporated into ethical, legal, and regulatory landscapes. From a cultural standpoint, this study adapts and explores psychometric properties of the Portuguese version of the Research Attitudes Questionnaire (RAQ), and analyzes the willingness of gamete donors and recipients to donate gametes and embryos for research and its association with sociodemographic, reproductive characteristics, and attitudes towards research. Between July 2017 and June 2018, 71 donors and 165 recipients completed a self-administered questionnaire at the Portuguese Public Bank of Gametes. Willingness to donate and attitudes towards research were measured with a 5-point Likert scale. RAQ psychometric characteristics were explored. Two RAQ components were identified: "trustworthiness of research" and "critical perspective". Most participants were willing to donate gametes and embryos: donors more willing to donate gametes and male recipients more willing to donate gametes and embryos. Higher RAQ scores, indicating a more positive attitude towards research, were observed on the component "trustworthiness of research" among those willing to donate gametes and embryos and on the component "critical perspective" among those willing to donate embryos. These findings help foster inclusivity, diversity, and responsiveness of research and call for upstream engagement of male and female gamete donors and recipients, promoting a trustworthy, anticipatory, democratic, and people-centered approach to policies, regulations, and practices in human gamete and embryo research.


Functional profiling of Covid 19 vaccine candidate by flow virometry.
Vaccine development is a complex process, starting with selection of a promising immunogen in the discovery phase, followed by process development in the preclinical phase, and later by clinical trials in tandem with process improvements and scale up. A large suite of analytical techniques is required to gain understanding of the vaccine candidate so that a relevant immunogen is selected and subsequently manufactured consistently throughout the lifespan of the product. For viral vaccines, successful immunogen production is contingent on its maintained antigenicity and/or infectivity, as well as the ability to characterize these qualities within the context of the process, formulation, and clinical performance. In this report we show the utility of flow virometry during preclinical development of a Covid 19 vaccine candidate based on SARS-CoV-2 spike (S) protein expressed on vesicular stomatitis virus (VSV). Using a panel of monoclonal antibodies, we were able to detect the S protein on the surface of the recombinant VSV virus, monitor the expression levels, detect differences in the antigen based on S protein sequence and after virus inactivation, and monitor S protein stability. Collectively, flow virometry provided important data that helped to guide preclinical development of this vaccine candidate.


In Situ-Formed and Low-Temperature-Deposited Nb:TiO2 Compact-Mesoporous Layer for Hysteresis-Less Perovskite Solar Cells with High Performance.
Recently, reported perovskite solar cells (PSCs) with high power conversion efficiency (PCE) are mostly based on mesoporous structures containing mesoporous titanium oxide (TiO2) which is the main factor to reduce the overall hysteresis. However, existing fabrication approaches for mesoporous TiO2 generally require a high-temperature annealing process. Moreover, there is still a long way to go for improvement in terms of increasing the electron conductivity and reducing the carrier recombination. Herein, a facile one-step, in situ, and low-temperature method was developed to prepare an Nb:TiO2 compact-mesoporous layer which served as both scaffold and electron transport layer (ETL) for PSCs. The Nb:TiO2 compact-mesoporous ETL-based PSCs exhibit suppressed hysteresis, which is attributed to the synergistic effect of the increased interface surface area caused by nano-pin morphology and the improved carrier transportation caused by Nb doping. Such a high-quality compact-mesoporous layer allows the PSCs assembled using optimized 2% Nb-doped TiO2 to achieve a remarkable PCE of 19.74%. This work promises an effective approach for creating hysteresis-less and high-efficiency PSCs based on compact-mesoporous structures with lower energy consumption and cost.


Single-Centre Retrospective Study Using Propensity Score Matching Comparing Left Versus Right Internal Jugular Vein Access for Transjugular Intrahepatic Portosystemic Shunt (TIPS) Creation.
To compare the safety and efficacy of left versus right internal jugular vein access for portal vein puncture during transjugular intrahepatic portosystemic shunt (TIPS) creation in patients with a small liver and short vertical puncture distance. The vertical distance from the hepatic vein orifice to the puncture point of the portal vein was measured by CT and DSA. A distance ≤ 30 mm is defined as a short vertical puncture distance. After 1:1 propensity score matching (PSM), 29 patients of left internal jugular vein-TIPS (LIJ-TIPS) and 29 patients of right internal jugular vein-TIPS (RIJ-TIPS) were included. The number of needle punctures, fluoroscopy time, and radiation dose during the puncture process were statistically analyzed. There was no significant difference in the average vertical puncture distances on CT or DSA between LIJ-TIPS and RIJ-TIPS (19.10 ± 0.60 mm vs. 19.30 ± 0.60 mm, P = 0.840; 22.02 ± 0.69 mm vs. 22.23 ± 0.64 mm, P = 0.822, respectively). The average number of needle punctures, fluoroscopy time, and radiation dose in LIJ-TIPS were significantly lower than those in RIJ-TIPS (2.07 ± 0.20 vs. 4.10 ± 0.24, P < 0.001; 78.45 ± 12.80 s vs. 201.16 ± 23.71 s, P < 0.001; 31.55 ± 7.04 mGy vs. 136.69 ± 16.38 mGy, P < 0.001, respectively). Within three punctures, the technical success rate in LIJ-TIPS was significantly higher than that in RIJ-TIPS (86.2 vs. 27.6%, P < 0.001). The incidence of hemoperitoneum in LIJ-TIPS was significantly lower than that in RIJ-TIPS (0% vs. 13.8%, P = 0.038). The left internal jugular vein could be used as primary access for TIPS creation in patients with a small liver and short vertical puncture distance.


Preliminary analysis of length and GC content variation in the ribosomal first internal transcribed spacer (ITS1) of marine animals.
Length and guanine-cytosine (GC) content of the ribosomal first internal transcribed spacer (ITS1) were compared across a wide variety of marine animal species, and its phylogenetic utility was investigated. From a total of 773 individuals representing 599 species, we only failed to amplify the ITS1 sequence from 87 individuals by polymerase chain reaction with universal ITS1 primers. No species was found to have an ITS1 region shorter than 100 bp. In general, the ITS1 sequences of vertebrates were longer (318 to 2,318 bp) and richer in GC content (56.8% to 78%) than those of invertebrates (117 to 1,613 bp and 35.8% to 71.3%, respectively). Specifically, gelatinous animals (Cnidaria and Ctenophora) were observed to have short ITS1 sequences (118 to 422 bp) with lower GC content (35.8% to 61.7%) than the other animal taxa. Mollusca and Crustacea were diverse groups with respect to ITS1 length, ranging from 108 to 1,118 and 182 to 1,613 bp, respectively. No universal relationship between length and GC content was observed. Our data indicated that ITS1 has a limited utility for phylogenetic analysis as obtaining confident sequence alignment was often impossible between different genera of the same family and even between congeneric species.


Impacts of nationwide lockdown due to COVID-19 outbreak on air quality in Bangladesh: a spatiotemporal analysis.
In Bangladesh, a nationwide lockdown was imposed on 26 March 2020, due to the COVID-19 pandemic. Due to restricted emissions, it was hypothesized that the air quality has been improved during lockdown throughout the country. The study is intended to assess the impact of nationwide lockdown measures on air quality in Bangladesh. We analyzed satellite data for four different air pollutants (NO2, SO2, CO, and O3) to assess the changes in the atmospheric concentrations of pollutants in major cities as well as across the country. In this study, the concentrations of NO2, SO2, CO, and O3 from 1 February to 30 May of the year 2019 and 2020 were analyzed. The average SO2 and NO2 concentrations were decreased by 43 and 40%, respectively, while tropospheric O3 were found to be increased with a maximum of > 7%. Among the major cities, Dhaka, Gazipur, Chattogram, and Narayanganj were found to be more influenced by the restricted emissions. In Dhaka, NO2 and SO2 concentrations were decreased approximately by 69 and 67%, respectively. Our analysis reveals that NO2 concentrations are highly correlated with the regional COVID-19 cases (r = 0.74). The study concludes that the lockdown measures significantly reduced air pollution because of reduced vehicular and industrial emissions in Bangladesh.


Impaired neutrophil store-mediated calcium entry in Type 2 diabetes.
In Type 2 diabetes impaired neutrophil function leads to increased bacterial infection and cardiovascular disease. Many neutrophil functions depend on calcium signalling, which involves release of calcium from intracellular stores and subsequently translocation of stores via the cytoskeleton to the plasma membrane, causing store-mediated calcium entry (SMCE) into the cell. We hypothesized that in Type 2 diabetes there would be a defect in SMCE. Neutrophils were prepared from patients with Type 2 diabetes (DM, n=15) and controls (NC, n=15). Free cytosolic calcium [Ca2+]i was measured with Fura-2 in resting cells and after stimulation of calcium release with fMLP and thapsigargin. Baseline [Ca2+]i was higher in neutrophils from the patients than the controls (NC 65 +/- 5 nm, DM 80 +/- 4 nm, P<0.05). However, after fMLP-treatment [Ca2+]i was significantly lower in the patients (NC 301 +/- 28 nm, DM 210 +/- 20 nm, P<0.01). The greater increase in controls was not observed when cells were treated with fMLP in the absence of extracellular calcium (-fold increase NC 2.9 +/- 0.5, DM 2.7 +/- 0.3). Treatment of cells with thapsigargin caused a similar greater increase in [Ca2+]i in the controls than in the patients that was not seen in the absence of extracellular calcium (-fold increase with Ca2+ NC 5.2 +/- 1.0, DM 3.0 +/- 0.4, P<0.05; fold increase without Ca2+ NC 2.5 +/- 0.4, DM 2.2 +/- 0.2). In Type 2 diabetes there is a defect in neutrophil calcium signalling which results in a lesser increase in free cytosolic calcium owing to impaired influx across the plasma membrane. Abnormal calcium signalling is likely to be important in the pathogenesis of diabetic complications.


Predictors of Increased Surgical Drain Output following Type I Thyroplasty for Glottic Insufficiency.
To determine predictors of increased drain output following type I thyroplasty for glottic insufficiency. Retrospective chart review. A retrospective review was conducted for patients who underwent type I thyroplasty for glottic insufficiency from 2014-2019. The primary outcome was 24-hour drain output. Increased drain output was defined as >50th percentile for the sample. Univariate logistic regression models and linear regression models were used. There were 84 patients with a mean age of 58.9 (SD 16.9) years. Twenty-four-hour drain output ranged from 0 to 29 mL with a mean of 9.47 (SD 6.49) mL. Patients with a history of tobacco use (OR 3.33; 95% CI, 1.24-8.95; P = .017) and prior neck surgery (OR 3.52; 95% CI, 1.26 to 9.83; P = .016) were significantly more likely to have increased drain output following surgery; these patients had a mean increase in 24-hour drain output of 3.51 mL (95% CI, 0.52 to 6.51; P = .022) and 1.74 mL (95% CI, -1.41 to 4.89; P = .274), respectively. Type of implant (Gore-Tex vs. Silastic; P = .425) and operative technique (unilateral vs. bilateral; P = .506) were not significantly associated with drain output. History of tobacco use and prior surgery of the neck predict increased drain output following type I thyroplasty surgery. These patients may derive the most benefit from surgical drain placement. More research is needed to confirm these findings and elucidate potential mechanisms. 4 Laryngoscope, 131:1557-1560, 2021.


Paradigm shifts in fungal secondary metabolite research.
The 8th International Mycological Congress (IMC8; Cairns, Australia) hosted several plenary lectures, poster presentations, and even entire symposia dedicated to fungal secondary metabolites (extrolites). These advances, presented in this special issue, together demonstrated how the impact of molecular biology and genomics and the availability of sophisticated methods of analytical chemistry has resulted in paradigm shifts in our understanding of fungal secondary metabolism and its key role in fungal biology. Rather than focus on classical topics such as discovery of novel drug candidates and identification of toxins, here we address two major themes in this special issue: (1) the utility and importance of secondary metabolites and their genes in polyphasic taxonomy, phylogeny, and evolutionary history of kingdom Fungi (syn. Eumycota); and (2) the genetic processes regulating secondary metabolite biosynthesis. The history of fungal chemotaxonomy and some important classes of secondary metabolites are reviewed.


Structure and phylogenetic diversity of post-fire ectomycorrhizal communities of maritime pine.
Environmental disturbances define the diversity and assemblage of species, affecting the functioning of ecosystems. Fire is a major disturbance of Mediterranean pine forests. Pines are highly dependent on the ectomycorrhizal (EM) fungal symbiosis, which is critical for tree recruitment under primary succession. To determine the effects of time since fire on the structure and recovery of EM fungal communities, we surveyed the young Pinus pinaster regenerate in three sites differing in the elapsed time after the last fire event. Pine roots were collected, and EM fungi characterized by sequencing the internal transcribed spacer (ITS) and the large subunit (LSU) regions of the nuclear ribosomal (nr)-DNA. The effects of the elapsed time after fire on the EM community structure (richness, presence/absence of fungi, phylogenetic diversity) and on soil properties were analysed.Fungal richness decreased with the elapsed time since the fire; although, the phylogenetic diversity of the EM community increased. Soil properties were different depending on the elapsed time after fire and particularly, the organic matter, carbon-to-nitrogen (C/N) ratio, nitrogen and iron significantly correlated with the assemblage of fungal species. Ascomycetes, particularly Tuberaceae and Pezizales, were significantly over-represented on saplings in the burned site. On seedlings, a significant over-representation of Rhizopogonaceae and Atheliaceae was observed in the most recently burned site, while other fungi (i.e. Cortinariaceae) were significantly under-represented. Our results are consistent with the hypothesis that fire can act as a selective agent by printing a phylogenetic signal on the EM fungal communities associated with naturally regenerated pines, pointing out to some groups as potential fire-adapted fungi.


Dissemination of Strongyloides stercoralis in a patient with systemic lupus erythematosus after initiation of albendazole: a case report.
Strongyloides stercoralis infection affects hundreds of millions of people worldwide. As immigration rates and international travel increase, so does the number of cases of strongyloidiasis in the United States. Although described both in immigrant and in immunosuppressed populations, hyperinfection and dissemination of S. stercoralis following the initiation of antiparasitic medication is a previously unreported phenomenon. Here we describe the case of a 38-year-old immunocompromised woman with systemic lupus erythematosus, who developed disseminated disease following treatment with albendazole (400 mg every 12 hours). Notably the patient was receiving oral prednisone (10 mg once daily), azathioprine (50 mg twice daily), and hydroxychloroquine (400 mg daily) at the time of hospitalization. The patient was subsequently treated successfully with ivermectin (200 mcg/kg daily). The reader should be aware that dissemination of S. stercoralis can occur even after the initiation of antiparasitic medication.


Chemical reconstruction of skin scars (CROSS) method for atrophic scars: A comprehensive review.
Chemical reconstruction of skin scars (CROSS) applies a high strength acid focally to treat atrophic scars. Although this method has gained popularity over the past two decades, no standardized treatment guideline exists for CROSS method in the treatment of atrophic scars. The purpose of this comprehensive review was to evaluate the indications, detailed techniques, efficacy, and safety of CROSS method. An extensive literature review was conducted to identify articles relating to CROSS method for atrophic scars from 2002 to 2018. The literature search yielded 19 articles meeting criteria. CROSS method has been used for the treatment of acne scars, varicella scars, enlarged pores, and depressed surgical scars. In studies using the quantile grading scale for acne scars, 60%-100% of patients showed >25% improvement. In two studies for varicella scars, 83%-100% of patients showed >25% improvement. CROSS method seems to be effective specifically for ice-pick scars. It is well tolerated and safe in Fitzpatrick skin phototypes I-V. Most reported complications are temporary and include postinflammatory dyspigmentation, erythema, pain, pruritus, infection, and widening of scars. This literature review suggests that CROSS method is a safe and effective treatment for atrophic scars, especially ice-pick scars, in skin types I-V. However, current published works have several limitations, including small sample sizes, lack of control group, different concentrations of acid, different frequency of treatments, and follow-up periods. Larger, randomized, controlled studies are needed to elucidate the optimal treatment protocol of CROSS method.


Stable isotope fractionation of cadmium in the soil-rice-human continuum.
Consumption of rice (Oryza sativa) grain is a major pathway by which humans are exposed to Cd, especially in non-smoking Asian populations. Although the stable isotope signatures of Cd offer a potential tool for tracing its sources, little is known about the isotopic fractionation of Cd across the entire soil-rice-human continuum. Cadmium isotope ratios were determined in field soils, rice grain, and human urine collected from two Cd-contaminated regions in southern China. Additionally, Cd isotopic fractionation in rice plants was investigated using two transgenic plants differing in Cd uptake and accumulation. Analysis of isotope ratios revealed a preferential enrichment of the heavy Cd isotopes from soil to rice grain (δ114/110Cdgrain-soil = +0.40‰) and from grain to urine (δ114/110Cdurine-grain = +0.40‰) in both regions. The first increase was mainly caused by partitioning between the soil solid phase and the soil solution, with heavier Cd preferentially enriching in the soil solution. Within the rice plant, we identified multiple processes that alter the isotope ratio, but the net effect throughout the plant was comparatively small. Cd fractionation in humans is presumably due to the preferential enrichment of heavier Cd isotopes by metal transporters DMT1 and ZIP8 (responsible for the absorption of Cd into body from the foods). These findings provide important insights into the Cd isotopic fractionation through the soil-rice-human continuum and are helpful for tracing the sources of Cd.


Spatiotemporal changes in summer days (SU25) in China from 1961 to 2017 and associated circulation factors.
Understanding the spatiotemporal variations in climate extremes indices, as well as the influencing factors, is critical to the scientific response to climate change. The temporal and spatial variations of SU25 (annual count of days when daily maximum temperature > 25 °C) were discussed in this study, based on daily maximum temperature data from 2398 meteorological stations in China from 1961 to 2017. The contributions of associated large-scale circulation factors to SU25 were quantitatively assessed by using the geographical detector method (GMD). The overall spatial distribution of SU25 was marked by a considerable increase from north to south. The SU25 increased significantly over time, with the national SU25 increasing at a rate of 2.5 days/decade. The Tibet Plateau (TP) had the slowest growth rate, with an average increase rate of 1.4 days/decade. The Hurst values of SU25 in all the subregions were generally high, indicating that most stations of SU25 would continue to increase in the future. Except for TP, the tipping years of other subregions were concentrated in the 1990s, and SU25 increased after the years. Among the large-scale circulation factors affecting SU25 in each subregion, Atlantic Multidecadal Oscillation (AMO) played a major role in SU25 variability. As a whole, the result of the pairwise interaction of each circulation factor was mainly nonlinear enhancement. The joint contributions of multiple factors to SU25 were larger than the contribution of each individual factor.


Clinical and genetic characterization of ten Egyptian patients with Wolf-Hirschhorn syndrome and review of literature.
Wolf-Hirschhorn syndrome (WHS) (OMIM 194190) is a multiple congenital anomalies/intellectual disability syndrome. It is caused by partial loss of genetic material from the distal portion of the short arm of chromosome. We studied the phenotype-genotype correlation. We present the clinical manifestations and cytogenetic results of 10 unrelated Egyptian patients with 4p deletions. Karyotyping, FISH and MLPA was performed for screening for microdeletion syndromes. Array CGH was done for two patients. All patients exhibited the cardinal clinical manifestation of WHS. FISH proved deletion of the specific WHS locus in all patients. MLPA detected microdeletion of the specific locus in two patients with normal karyotypes, while array CGH, performed for two patients, has delineated the extent of the deleted segments and the involved genes. LETM1, the main candidate gene for the seizure phenotype, was found deleted in the two patients tested by array CGH; nevertheless, one of them did not manifest seizures. The study emphasized the previous. WHS is a contiguous gene syndrome resulting from hemizygosity of the terminal 2 Mb of 4p16.3 region. The Branchial fistula, detected in one of our patients is a new finding that, to our knowledge, was not reported.


There's plenty of light at the bottom: statistics of photon penetration depth in random media.
We propose a comprehensive statistical approach describing the penetration depth of light in random media. The presented theory exploits the concept of probability density function f(z|ρ, t) for the maximum depth reached by the photons that are eventually re-emitted from the surface of the medium at distance ρ and time t. Analytical formulas for f, for the mean maximum depth 〈zmax〉 and for the mean average depth reached by the detected photons at the surface of a diffusive slab are derived within the framework of the diffusion approximation to the radiative transfer equation, both in the time domain and the continuous wave domain. Validation of the theory by means of comparisons with Monte Carlo simulations is also presented. The results are of interest for many research fields such as biomedical optics, advanced microscopy and disordered photonics.


Factors affecting PBSC mobilization and collection in healthy donors.
Peripheral blood stem cells are widely used as stem cell source for allografting. Progenitor cells can be effectively mobilized into peripheral blood in majority of healthy donors with a brief administration of G-CSF. A mobilization course in 111 donors (median age 40years) was retrospectively studied and the factors influencing the efficacy of mobilization were analyzed. The median number of CD34+ cells per kg recipient weight 5.1x10(6) was obtained after a median of two aphereses. The target cell dose (4.0x10(6)/kg) was reached in 69% of donors. Circulating CD34+ count and CD34+ yield were negatively associated with donor's age. Other independent factors associated with superior yield were precollection platelet and WBC counts. In multivariate analysis only CD34+ precount predicted for CD34+ yield. G-CSF had an acceptable short-term safety profile. Our data confirm that apheresis is a safe procedure in healthy including aged donors and suggest that older donors could be poorer mobilizers than younger.


Arteriovenous fistulas at the cervicomedullary junction presenting with subarachnoid hemorrhage: six case reports with special reference to the angiographic pattern of venous drainage.
Cases with spinal perimedullary arteriovenous fistulas (SPAVFs) or spinal dural arteriovenous fistulas (SDAVFs) at the cervicomedullary junction are rare. We performed a retrospective, angiographic study of 6 such patients to assess whether available angiographic data were predictive of the risk for hemorrhage. We report 6 patients with arteriovenous fistulas at the cervicomedullary junction. All presented with subarachnoid hemorrhage (SAH). Angiography demonstrated that 4 of the 6 fistulas were SDAVFs fed by the meningeal branch of the vertebral artery; the other 2 were SPAVFs fed by the anterior spinal artery. Drainage was via the perimedullary vein of the cervicomedullary junction. An ascending venous route into the intracranial sinus was recognized in all 6 cases; in 3 the draining system contained varices. In 2 cases, the venous route was on the ventral side of the brain stem with drainage into the cavernous sinus. In 4 cases, the venous route was lateral at the brain stem with drainage into the inferior petrosal sinus. SPAVFs and SDAVFs at the cervicomedullary junction that manifest an ascending venous route into the intracranial sinus present an increased risk for SAH.


Association Between Oral Anticoagulants and Stroke Severity at Onset in Elderly Patients with Cardioembolic Stroke Due to Non-Valvular Atrial Fibrillation.
This study aimed to determine whether oral anticoagulant therapy affects the severity of cerebral infarction at onset in elderly patients with non-valvular atrial fibrillation. This retrospective study included 330 elderly patients (aged ≥75 years) who were hospitalized for cardioembolic stroke due to non-valvular atrial fibrillation. Patients' medical history, stroke severity at onset (National Institutes of Health Stroke Scale score), and the prevalence of large vessel occlusion were compared between patients who received oral anticoagulant therapy (n = 109) and those who did not receive oral anticoagulant therapy (n = 221). Stroke severity was significantly lower in patients who received anticoagulants than in those who did not receive anticoagulants (6 versus 12; P = 0.021). Patients who did not receive anticoagulants had a significantly higher prevalence of large vessel occlusion (52% versus 37%; P = 0.010). After resampling based on propensity score matching, both median stroke severity (7 versus 12; P = 0.046) and large vessel occlusion prevalence (36% versus 57%; P = 0.019) were significantly lower in patients who received anticoagulant therapy. The results of this study suggest that elderly patients with non-valvular atrial fibrillation who are administered oral anticoagulant therapy before the onset of cerebral infarction develop less severe stroke than those who are not receiving oral anticoagulant therapy. Thus, oral anticoagulant therapy should be actively considered in patients with non-valvular atrial fibrillation as it does not only prevents cerebral embolism, but also reduces the risk of severe sequelae.


TIGIT Blockade and IL15 in Tumor Immunotherapy: Together is Better.
IL15 and TIGIT blockade enhances the natural killer (NK) cell-mediated activity against MHC-I-deficient melanoma both in vitro and in preclinical models. IL15-induced harnessing of NK cells, associated with their unleashing by TIGIT blockade, may represent a therapeutic approach for tumors, which lack HLA-I molecules thus escaping the CD8+ T cell-mediated control.See related article by Chauvin et al., p. 5520.


Expression of a Fas-like proapoptotic molecule on the lymphocytes of Xenopus laevis.
Ligation of the externally expressed Fas (APO1/CD95) molecule will initiate programmed cell death (apoptosis), in many mammalian developing and adult cells. Fas-induced apoptosis has not been demonstrated with the cells of any non-mammalian vertebrate. We immunostained suspensions of splenocytes from adult Xenopus laevis, the South African clawed toad, with a polyclonal rabbit anti-human Fas antibody raised against the amino acid residues 321-335 of human Fas. The binding was specific, as it was dramatically reduced by preincubation of the antibody with the Fas peptide used to make it, but not with a Fas-ligand (FasL) peptide. The binding was enhanced after in vitro exposure of the splenocytes to phytahemagglutinin (PHA), a T cell mitogen and apoptogen in this species. Sections of developing Xenopus larval tissue were also immunostained with the polyclonal rabbit anti-human Fas antibody. Consistent binding of thymocytes and splenocytes was not observed until early metamorphosis in these immunological sites. A monoclonal mouse anti-human Fas antibody, previously used to stimulate apoptosis in mammalian cells, induced significant levels of apoptosis in adult Xenopus splenocytes and additionally, bound specifically to a splenocyte extract, as assayed by ELISA. Thus, a molecule on Xenopus splenocytes shares both structural and functional homologies with human Fas, indicating the evolutionary conservation within vertebrates of this means of initiating apoptosis.


The number of involved extracranial organs: a new predictor of survival in breast cancer patients with brain metastasis.
This study was performed to investigate the potential impact of the number of involved extracranial organs on survival in patients with brain metastasis from breast cancer. The data of 196 patients treated with whole-brain radiotherapy (WBRT) alone for brain metastases from breast cancer were retrospectively analyzed. Six potential prognostic factors were evaluated for associations with survival. These factors included WBRT regimen, age, Karnofsky performance score (KPS), number of brain metastases, interval from breast cancer diagnosis to WBRT, and the number of involved extracranial organs. The 6-month survival rates of patients with involvement of 0, 1, 2, 3 and ≥4 extracranial organs were 59%, 49%, 26%, 26% and 13%, respectively, and the 12-month survival rates were 45%, 36%, 17%, 17% and 13%, respectively (p<0.001). On multivariate analysis, the number of involved extracranial organs (risk ratio 1.17; 95%-confidence interval 1.02-1.35; p=0.028) maintained significance, as did KPS (p<0.001), but not age (p=0.27). The number of involved extracranial organs is an independent prognostic factor of survival in patients with brain metastasis from breast cancer.


Cataract prevalence following a nationwide policy to shorten wait time for cataract surgery.
Cataract is an age-related eye disease. Visual impairment from cataract can be restored by cataract surgery. In 2004 the Canadian federal government invested in a multibillion dollar wait time strategy to shorten the wait time for cataract surgery, a government-insured health service in all Canadian jurisdictions. We assessed if this nationwide policy reduced the number of Canadians waiting for cataract surgery as more individuals with cataract were free of cataract following the rapidly conducted surgery. In this cross-sectional study we analyzed data from randomly selected individuals aged ≥ 45 years responding to the Canadian Community Health Survey (CCHS) in 2000/2001, 2003, 2005, and the CCHS Healthy Aging in 2008/2009. Information on cataract was obtained from self-reported questionnaire. The age- and sex-standardized prevalence of cataract was calculated for comparisons. Cataract was reported by 0.93 million Canadians in 2000/2001, 0.99 million in 2003, 1.10 million in 2005, and 1.34 million in 2008/2009. This corresponds to an age- and sex-standardized prevalence of 8.9% in 2000/2001, 9.0% in 2003, 9.5% in 2005, and 10.2% (P <0.05) in 2008/2009. The increase in age- and sex-standardized prevalence was greater in individuals without secondary school graduation than those with secondary school graduation or higher (4.3% versus 1.3%, P < 0.05) and was seen in all Canadian provinces. The largest increase was documented in a province (Saskatchewan, from 9.8% in 2000/2001 to 12.6% in 2008/2009, P < 0.05) with the longest median wait times for cataract surgery (118 days in 2008) and the lowest number of ophthalmologists per 100 000 population (1.96 versus 3.35 national average). The age- and sex-standardized prevalence of cataract increased 4‒5 years after the multibil- lion-dollar wait time strategy was launched in 2004. A lower threshold to diagnose cataract may be one potential reason for this finding. Further research is needed to understand the true reasons for the increase.


Effects of hyperthermia on the peripheral metabolism of ammonia and glutamine.
Although there are a number of physiological states associated with an increase in body temperature, eg, nonseptic and septic fever or exercise, the effects of hyperthermia on intermediary metabolism in vivo have received little attention. The current study was undertaken to examine the peripheral metabolism of ammonia and ammonia-related amino acids using a model of exogenously induced mild hyperthermia in nonfebrile subjects. Arteriovenous levels of metabolites were measured in subjects with known cerebrovascular insufficiency. Peripheral exchange of these metabolites was monitored before, during, and after hyperthermia to an average temperature of 38.5 degrees C. All subjects showed an increase in plasma ammonia and glutamate. There was also an associated peripheral production of ammonia and an uptake of glutamine and alanine by the skeletal muscle tissues. The exchange of glutamate was not affected. These findings indicate that hyperthermia is associated with a nitrogen-sparing effect, the mechanism of which is yet unknown.


Unusual cause of recurrent arterial embolism: floating thrombus in the aortic arch surgically removed under hypothermic cardiocirculatory arrest.
A 46-year-old fully active, asymptomatic man suffered two episodes of major peripheral arterial embolism within 2 months. Heart disease was ruled out by appropriate investigations. Further diagnostic evaluation (angiography, CAT scan) revealed the extremely rare finding of a "floating mass" in the transverse aortic arch suspected to be the source of embolization. This mass was successfully removed using the technique of hypothermic cardiocirculatory arrest. The histological diagnosis was an aged intraluminal thrombus and moderate atherosclerosis of the thoracic aorta. For prevention of recurrent arterial embolism in cases without an initially apparent cause and site of origin, a thorough diagnostic, and in a given patient, an aggressive surgical approach for the elimination of the embolic source are advocated.


"Descemet Membrane Detachment": A Novel Concept in Diagnosis and Classification.
To examine the optical coherence tomography (OCT) and histologic features of Descemet membrane detachment (DMD) to ascertain the involvement of the pre-Descemet layer (PDL). Retrospective, observational case series. Clinical, histopathologic, and OCT features of a cohort of 41 cases with diagnosis of DMD from 4 centers were studied. OCT images were evaluated independently by 3 observers for number of detached layers (1 or 2), reflectivity, configuration (straight line or wavy), distance from posterior stroma, and presence or absence of a tear with any scrolling of the torn edges. Five had a histology specimen. The main outcome measure was the involvement of the PDL in DMD and its confirmation by histology. Three types of DMD were identified: type 1, where the PDL and DM were detached together; type 2, where only the DM was detached; and mixed, where the PDL and DM were detached but also separated from each other. These were further found to be rhegmatogenous or nonrhegmatogenous depending on the presence of absence of a tear in DM or both layers. Histology confirmed involvement of PDL in all 5 cases and showed it to be infiltrated by cells in 3 of 5 cases. The PDL is involved in DMD. This fact significantly changes our understanding of DMD and could have implications for management. The detached PDL can be infiltrated with cells. A prospective study in relation to etiology and types of DMD is needed.


I-PV: a CIRCOS module for interactive protein sequence visualization.
Today's genome browsers and protein databanks supply vast amounts of information about proteins. The challenge is to concisely bring together this information in an interactive and easy to generate format. We have developed an interactive CIRCOS module called i-PV to visualize user supplied protein sequence, conservation and SNV data in a live presentable format. I-PV can be downloaded from http://www.i-pv.org. firstname.lastname@example.com, firstname.lastname@example.com or firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


The impact of the COVID-19 pandemic on ICU clinical trials: a description of one research team's experience.
Disruptions to clinical trials conducted in the intensive care unit (ICU) due to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV2; coronavirus disease 2019 [COVID-19]) pandemic included fewer new trials activated and more trials stopped. While a number of ongoing, non-COVID-19 clinical trials remained open to enrollment, the direct impact of the pandemic on ICUs instilled chaos in this already challenging environment. The numerous challenges need to be reported so investigators can proactively plan and manage these myriad challenges. Thus, the purpose of this study was to describe the impact of the COVID-19 pandemic on screening and accrual for a non-COVID-19 parent clinical trial enrolling critically ill ICU patients receiving mechanical ventilatory support. A descriptive, retrospective design using quantitative data from detailed screening logs and qualitative observations with field notes from a parent clinical trial were used to address the objectives. The primary aims of the two-site parent clinical trial (n = 190) are to test the efficacy of self-administration of sedative therapy by mechanically ventilated ICU patients on anxiety and delirium occurrence. ICUs from two academic medical centers [names removed for blinding] plus a community hospital in Minnesota were screened daily for alert patients (Richmond Agitation Sedation Scale [RASS] - 2 to + 1), following commands, hemodynamically stable with sufficient hand grip strength to depress a push-button device. Screening data were summarized based on the primary reason patients were not enrolled (screen failures, declinations of consent). Descriptive statistics (frequencies, percentages), chi-square, and Fisher's Exact test were used to describe the data and to determine any differences among distributions of screening failures and recruitment declinations during the defined pre-pandemic (August 27, 2018-March 15, 2020, 2976 screened patients) and pandemic timeframes (March 16, 2020-February 28, 2022, 3912 screened patients). Qualitative data from varied sources such as screening logs, institutional email communications, staff field notes, and research team meeting minutes were summarized into themes. Despite significantly fewer screen failures due to hypotension, cognitive impairment/dementia, coma, or chemical paralysis with 938 additional patients on the screening log, more were accrued pre-pandemic (n = 55) than during the pandemic period (n = 45); declination reasons were non-significant. Pandemic burdens experienced by study personnel, ICU care providers, and patients/families were revealed that attributed to decreased accrual. While the parent clinical trial remained opened, cumulative factors adversely impacted the trial during the pandemic period with fewer patients accrued. The human toll of the pandemic on research staff, clinicians, and patients/family members demands that investigators be proactive in managing these challenges to conduct ICU clinical trials successfully, including careful oversight of human and financial resources. ClinicalTrials.gov NCT#02,819,141 Registered 29 June 2016.


Air Medical Curricula in Emergency Medicine Residencies: A National Survey.
The aim of this study was to determine if a common air medical curriculum exists among Accreditation Council for Graduate Medical Education (ACGME) emergency medicine (EM) residencies. Institutional review board exemption was obtained. A cross-sectional survey study design was used. All ACGME-approved EM residencies as of February 2013 were identified, and the program directors were e-mailed a 3-question survey. Descriptive statistics were calculated for each response, and a response bias analysis was completed. All 160 ACGME EM residencies were contacted by e-mail. One hundred six (66%) programs responded. Sixty-nine (65%) of the respondents offered an air medical experience. Only 25 of the 69 (36%) stated that they had a formal air medical curriculum, and 15 programs provided a copy of their curriculum. Protocol education was present in 80% of programs. Safety training was present in 60% of the programs. Financial education was present in 7% of programs. There was no statistically significant difference between responders and nonresponders except for residency class size. After 30 years of published articles and position statements calling for standardized air medical resident crew member training, there is still no standardized training program among emergency medicine residencies.


"Hands off hand hygiene training": Implementation of a COVID safe auditor training program.
Optimal hand hygiene practices reduce the risk of healthcare-associated infections, especially in high-risk settings of immunocompromised patients. In 2020, face-to-face learning was disallowed in the environment of coronavirus disease 2019 transmission. We developed a revised learning program for hand hygiene auditors for our cancer care facility. The learning package resulted in a 2-fold increase in the number of participants, with effective promotion by managers, due in part to reduced time and resources for training, and flexibility for staff.


A mass merchandiser's role in enhancing pharmacy students' business plan development skills for medication therapy management services.
To develop a relationship between a pharmacy management course and a mass merchandiser and to determine whether involving pharmacy managers from the mass merchandiser in the course would enhance student skills in developing a business plan for medication therapy management services. The pharmacy managers from the mass merchandiser participated in lectures, provided panel discussions, and conducted a business plan competition. Learning was assessed by means of 4 examinations and 1 project (ie, the business plan). At the conclusion of the semester, surveys were administered to solicit student input and gain insight from pharmacy managers on the perceived value of this portion of the course. Students' average grade on the business plan assignment, which included the oral presentation, the peer assessment, and the written proposal, was 92.2%. Approximately 60% (n=53) of surveyed students agreed or strongly agreed that their management skills had improved because of the participation of pharmacy managers from the mass merchandiser. All of the managers enjoyed participating in the experience. The involvement of pharmacy managers from a mass merchandiser enhanced student learning in the classroom, and managers felt that their participation was an important contribution to the development of future pharmacists.


miR-425 inhibits melanoma metastasis through repression of PI3K-Akt pathway by targeting IGF-1.
miR-425 is a potential tumor suppressor in cancer, but its role in melanoma is still unknown. We aim to investigate miR-425 expression in melanoma tissues and cell lines. Next, cell proliferation, cell cycle, apoptosis and metastasis will be studied using lentivirus-mediated gain-of-function studies. The predicted results are stable miR-425 inhibits cell proliferation and metastasis and induced cell apoptosis. It is predicted that IGF-1 is a potential target gene of miR-495 by bioinformatics analysis. Then luciferase assay analysis identifies IGF-1 as a new direct target gene of miR-425 and miR-425 inhibits melanoma cancer progression via IGF-1. Collectively, our findings suggested that miR-425 may function as a tumor suppressor in melanoma by targeting IGF-1.


The Assessment of Depression Inventory (ADI): an appraisal of validity in an inpatient sample.
This study examined the validity of the Assessment of Depression Inventory (ADI), a 39-item measure of depression with three embedded validity scales, using inpatient participants. The ADI Depression (Dep) and Feigning (Fg) scales were compared to the corresponding scales of a widely used multiscale instrument, the Personality Assessment Inventory (PAI). The correlations between this test and the ADI appear to indicate that the ADI is effective in measuring the constructs of both depression and purposeful distortion. The efficacy of various ADI Fg scale cut scores was addressed using the PAI validity scales as a criterion. A receiver operator curve based on this data produced an area under the curve of 85.3% for 14, the most optimal cut score.


Epileptiform activity induced by low Mg2+ in cultured rat hippocampal slices.
Organotypic cultured slices of the rat hippocampus undergo synaptic reorganization. Besides the establishment of reciprocal connections between area CA1 and the dentate gyrus (DG), collateral excitatory connections between granule cells are formed which are similar to those appearing in several epilepsy models and in the DG from patients with temporal lobe epilepsy. We studied the characteristics of epileptiform activity induced by low Mg2+ perfusion in cultured hippocampal slices using extra- and intracellular recordings. With low Mg2+ perfusion synchronous seizure like events (SLEs) were readily observed in the DG and areas CA3 and CA1. Also, the isolated DG was able to display seizure like activity. Intracellular recordings revealed long lasting depolarization shifts in granule cells of the DG and pyramidal cells of areas CA3 and CA1. The SLEs, lasting 2-3 s, could be recorded for at least 3 h in areas CA1 and CA3. However, approximately an hour after perfusion with low Mg2+, the epileptiform activity disappeared in the DG and responses to single pulse hilar stimulation progressively deteriorated. These responses returned to control values 1 week after reincubating the cultures. Interestingly, no deterioration of stimulus induced responses was observed in the isolated DG after exposure to low Mg2+.


Breast cancer with chest wall progression: treatment with photodynamic therapy.
Chest wall progression of breast carcinoma affects up to 5% of breast cancer patients and is a major source of their pain. Treatment options are limited or may not be offered to these patients. Low-dose Photofrin-induced photodynamic therapy (PDT) offers an excellent clinical response with minimal morbidity. We report our continued experience with PDT in this setting. Fourteen patients with more than 500 truncal metastases were treated with PDT. All received off-label Photofrin (.8 mg/kg) i.v. and light treatment at 630 nm from a diode laser with a microlens at a fluence of 1800 mW and a total light dose of 150 to 200 J/cm2 at 48 hours. One patient required re-treatment because of extensive disease. Follow-up was at least 6 months, and several extended to >24 months. All patients demonstrated tumor necrosis, with 9 of 14 complete responses, including with lesions >2 cm in thickness. Disease progression occurred outside of the treatment field. Several patients had initial regression of untreated lesions. Wound care, especially with disease in the deep tissues, was an issue. Low-dose Photofrin-induced PDT offers patients with chest wall progression a treatment option with an excellent clinical response. To date, the response is prolonged and offers good local control. Surgical oncologists have an active role in this treatment option.


Functional and molecular genetic analysis of a group of rat class I major histocompatibility genes which might be homologous to H-2Qa.
Studies on the major histocompatibility complex of the rat (RT1) will be summarized, which demonstrate the existence of a cluster of class I-like genes differing in expression, function and complexity of their organization from the "classical" class I genes, the gene products of which restrict antigen recognition by T lymphocytes. The characterization of the respective gene region (RT1.C) and its products is based on data obtained with classical genetic, molecular genetic, biochemical and immunological methods. It turns out that the RT1.C region and RT1.C antigens are in many aspects homologous to the H-2Qa region and H-2Qa antigens, respectively, of the mouse.


Protective role of cell division cycle 48 (CDC48) protein against neurodegeneration via ubiquitin-proteasome system dysfunction during zebrafish development.
Cell division cycle 48 (CDC48), a ubiquitin-dependent molecular chaperone, is thought to mediate a variety of degradative and regulatory processes and maintain cellular homoeostasis. To investigate the protective function of CDC48 against accumulated ubiquitinated proteins during neurodevelopment, we developed an in vivo bioassay technique that detects expression and accumulation of fluorescent proteins with a polyubiquitination signal at the N terminus. When we introduced CDC48 antisense morpholino oligonucleotides into zebrafish embryos, the morphant embryos were lethal and showed defects in neuronal outgrowth and neurodegeneration, and polyubiquitinated fluorescent proteins accumulated in the inner plexiform and ganglion cell layers, as well as the diencephalon and mesencephalon, indicating that the degradation of polyubiquitinated proteins by the ubiquitin-proteasome system was blocked. These abnormal phenotypes in the morphant were rescued by CDC48 or human valosin-containing protein overexpression. Therefore, the protective function of CDC48 is essential for neurodevelopment.


Determination of the four major surfactant classes in cleaning products by reversed-phase liquid chromatography using serially connected UV and evaporative light-scattering detection.
A method for the simultaneous determination of the most frequently used surfactant families -linear alkyl benzenesulphonates (LAS), alkyl ether sulphates (AES), fatty alcohol ethoxylates (FAE) and oleins (soaps, fatty acid salts) - in cleaning products, has been developed. The common reversed phase octyl (C8), pentafluorophenyl and biphenyl columns were not capable of separating the anionic LAS and AES classes; however, since only LAS absorbs in the UV, these two classes were independently quantified using a C8 column and serially connected UV and ELSD detection. The best compromise to resolve the four surfactant classes and the oligomers within the classes was achieved with a C8 column and an ACN/water gradient. To enhance retention of the anionic surfactants, ammonium acetate, as an ion-pairing agent compatible with ELSD detection, was used. Also, to shift the olein peaks with respect to that of the FAE oligomers, acetic acid was used. In the optimized method, modulation of the mobile phase, using ammonium acetate during elution of LAS and AES, and acetic acid after elution of LAS and AES, was provided. Quantitation of the overlapped LAS and AES classes was achieved by using the UV detector to quantitate LAS and the ELSD to determine AES by difference. Accuracy in the determination of AES was achieved by using a quadratic model, and by correcting the predicted AES concentration according to the LAS concentration previously established using the UV chromatogram. Another approach also leading to accurate predictions of the AES concentration was to increase the AES concentrations in the samples by adding a standard solution. In the samples reinforced with AES, correction of the predicted AES concentration was not required. FAE and olein were quantified using also quadratic calibration.


NF-κB as a regulator of cancer metastasis and therapy response: A focus on epithelial-mesenchymal transition.
Metastasis of tumor cells is a complex challenge and significantly diminishes the overall survival and prognosis of cancer patients. The epithelial-to-mesenchymal transition (EMT) is a well-known mechanism responsible for the invasiveness of tumor cells. A number of molecular pathways can regulate the EMT mechanism in cancer cells and nuclear factor-kappaB (NF-κB) is one of them. The nuclear translocation of NF-κB p65 can induce the transcription of several genes involved in EMT induction. The present review describes NF-κB and EMT interaction in cancer cells and their association in cancer progression. Due to the oncogenic role NF-κB signaling, its activation enhances metastasis of tumor cells via EMT induction. This has been confirmed in various cancers including brain, breast, lung and gastric cancers, among others. The ZEB1/2, transforming growth factor-β, and Slug as inducers of EMT undergo upregulation by NF-κB to promote metastasis of tumor cells. After EMT induction driven by NF-κB, a significant decrease occurs in E-cadherin levels, while N-cadherin and vimentin levels undergo an increase. The noncoding RNAs can potentially also function as upstream mediators and modulate NF-κB/EMT axis in cancers. Moreover, NF-κB/EMT axis is involved in mediating drug resistance in tumor cells. Thus, suppressing NF-κB/EMT axis can also promote the sensitivity of cancer cells to chemotherapeutic agents.


Identifying a sample of elderly people by a postal screen.
This paper describes how a sample of elderly people were identified from the electoral register using an initial postal screen, and reports a technique that was used to increase the response to the screen.