Analytical techniques used to study the degradation of proteins and peptides: chemical instability.
Instability of peptides and proteins can be divided into two forms: chemical and physical instability. Chemical instability is due to modification/alteration of amino acid residues. There are several types of degradation reactions responsible for this instability. Most frequently described reactions are oxidation, reduction, deamidation, hydrolysis, arginine conversion, beta-elimination and racemisation. However, any study of the degradation of a chemical substance lacks reliability when the analytical methodology, that is used is not properly validated. Especially in the investigation, where degradation processes lead to their parent compounds, validation of the analysis is pivotal for the correct interpretation of the results. It is therefore appropriate and useful to assemble an overview of degradation processes in relation to the analytical methods to monitor them. An overview like this can help investigators to make the right choices in their analytical approach of stability problems. The degradation reactions involved in peptide/protein degradation as well as the methods to monitor them are summarized and discussed.


Isolation and characterization of culturable endophytic bacterial community of stripe rust-resistant and stripe rust-susceptible Pakistani wheat cultivars.
In this study, endophytic bacteria isolated from root, stem, and leaf tissues of stripe rust-susceptible (Inqilab 91, Galaxy 2013, and 15BT023) and stripe rust-resistant (NARC 2011, Ujala 2015, TW1410) cultivars were identified and characterized. Abundance of endophytes was found in roots as compared with stems and leaves. Resistant and susceptible cultivars significantly differed in abundance of endophytic bacteria. Restriction analysis of 16S rRNA genes amplified from 100 bacterial isolates produced 17 unique patterns. Representatives of each of the 17 unique patterns were sequenced and identified. Among the sequenced bacteria, 8 belonged to Firmicutes, 7 were Proteobacteria, and 2 were Actinobacteria. Most of the isolates have plant growth-promoting properties and a few have the potential of producing hydrolytic enzymes. Two isolates showed significant inhibition of rust spore germination. These endophytic bacteria not only can be helpful in growth-promoting activities but also can assist in biocontrol of stripe rust disease.


Factors affecting nursing students' incivility: As perceived by students and faculty staff.
Students' incivility in institutions of higher education is a serious issue that faces educators in performing their teaching duties. The negative impacts associated with uncivil classroom behaviors have been found to contribute to the disruption of the learning process and the classroom learning environment, and the deterioration of the faculty-student relationship. This study assays the incivility level among nursing students, investigates factors affecting student nurses' incivility, and explores the relationship between students' uncivil behavior and factors affecting its occurrence based on the perceptions of students and faculty staff. A descriptive comparative research design included all nursing students (n=186) and faculty staff (n=66) in the Faculty of Nursing, Port Said University. A structured questionnaire was used to collect the data. The results of the study reflected that less than two thirds of students (60.2%) reported irresponsible behaviors, more than half (55.9%) expressed that they behave inappropriately, and 47.8% of them believed that they behave aggressively. The highest percentage of students (55.4%) recorded a high level of uncivil behavior, while faculty staff recorded a lower level regarding aggressive uncivil student behaviors. Both faculty staff and students agreed that a high level of incivility is affected by the studied factors, including issues related to environmental and study climate, faculty policies, political atmosphere, and faculty staff. Uncivil students' behavior interferes with academic achievement and leads to a declined curve of ethics for nursing students, who are to be considered a symbol of ethics when dealing with their patients. Based on the study results, activated implementation of faculty policies on uncivil behaviors is recommended. Also, there is an obvious need to train faculty staff members to deal with uncivil and bullying students.


Denonvilliers' fascia in men: a sheet plastination and confocal microscopy study of the prerectal space and the presence of an optimal anterior plane when mobilizing the rectum for cancer.
The aim of this study was to investigate the detailed, in situ, morphology of Denonvilliers fascia (DVF) in cadavers using sheet plastination and confocal microscopy and to review and describe the optimal anterior plane for mobilisation of the distal rectum.. Six, male cadavers (age range, 46-87 years) were prepared as six sets of transverse (x2), coronal (x1) and sagittal (x3) plastinated sections which were examined under a confocal laser scanning microscope. In this study a consistent space between the anterior rectal wall and the posterior surface of the prostate and seminal vesicles above the level of the perineal body was termed the prerectal space. Within that prerectal space we identified fibres which take their origin from the external urethral sphincter (EUS), together with others from the longitudinal rectal muscle (LRM) and the connective tissue sheaths of neurovascular bundles. Neither the EUS- nor the LRM-originated fibres were continuous with the endopelvic fascia;they are interposed laterally and cranially by multiple neurovascular bundles. Further, our results suggest that the peritoneum does not descend deep within the prerectal space. This study reveals the undisturbed, in situ, structural detail of membrane-like structures in the prerectal space and confirms that the optimal plane for anterolateral mobilization of the rectum is posterior to the multilayered DVF. This article is protected by copyright. All rights reserved.


A new indole glycoside from Kitasatospora sp. MG372-hF19 carrying a 6-deoxy-α-L-talopyranose moiety.
Small cell lung cancer (SCLC) is a severe malignancy with early and widespread metastasis, and novel therapeutic drugs are needed. To identify cytotoxic natural compounds against SCLC, we screened libraries of microbial fermentation broths using several lung cancer cell lines. We found that the actinomycete strain MG372-hF19 produces a compound that has not been isolated from natural sources but previously chemically synthesized, 6-chloro-1H-indole-3-carboxaldehyde (1), and an entirely new compound, named 6-deoxy-α-L-talopyranose 1-(6-chloro-1H-indole-3-carboxylate) (2), together with leptomycins. The molecular formulas of the compounds were established as C9H6ClNO and C15H16ClNO6, respectively, via high-resolution electrospray ionization mass spectrometry, and their structures were determined using detailed NMR. Absolute configurational analysis of the sugar unit of compound 2 revealed that the compound incorporates the rare deoxyhexose 6-deoxy-α-L-talopyranose. Both compounds exhibited weak growth-inhibiting activities against human lung cancer cell lines.


Tertiary trisomy 14q--, due to paternal balanced translocation 46,XY,t(1;14)(q44;q22).
Retardation of growth and mental development, craniofacial dysmorphy, limb anomalies, cryptorchidism and repeated infections are observed in a child with 47,XY,+der(14),t(1;14)(q44;q22)pat.


Liver cadmium levels in North Carolina residents who died of heart disease.
Using data from an autopsy series, we found a strong positive correlation between liver concentration of cadmium and death from heart disease. In view of recent experiments reported in the literature, the possibility that a low level of cadmium has a toxic effect on the cardiac conduction system is supported.


Similarity of protein conformation at low pH and high temperature observed for B-chains of two plant toxins: ricin and mistletoe lectin 1.
A comparative study of subunits of two plant toxins, ricin (RC) and mistletoe lectin 1 (ML 1), has been undertaken. The study suggests that isolated B-chains of these toxins undergo structural transitions at low pH (from 5 to 4) and high temperature (45 degrees C), and as a result of the guanidine hydrochloride denaturing effect (to 3 M). Our results indicate that the protein conformation observed at low pH and high temperature are similar, though not identical. These conformations differ from the native one (pH 7, 25 degrees C), the protein in these conformations has a low fluorescence tryptophan intensity, and tryptophans are more exposed to aqueous solutions. However, these conformations differ also from the state unfolded by guanidine hydrochloride. An assumption is made that the partially denatured protein structure, exhibited at low pH and high temperature, is a functionally essential intermediate state of the toxin B-chain.


Chronically lowering sympathetic activity protects sympathetic nerves in spleens from aging F344 rats.
In the present study, we investigated how increased sympathetic tone during middle-age affects the splenic sympathetic neurotransmission. Fifteen-month-old (M) F344 rats received rilmenidine (0, 0.5 or 1.5mg/kg/day, i.p. for 90 days) to lower sympathetic tone. Controls for age were untreated 3 or 18M rats. We report that rilmenidine (1) reduced plasma and splenic norepinephrine concentrations and splenic norepinephrine turnover, and partially reversed the sympathetic nerve loss; and (2) increased β-adrenergic receptor (β-AR) density and β-AR-stimulated cAMP production. Collectively, these findings suggest a protective effect of lowering sympathetic tone on sympathetic nerve integrity, and enhanced sympathetic neurotransmission in secondary immune organs.


Sucrose absorption by the rat small intestine in vivo and in vitro.
1. The absorption of glucose and fructose derived from sucrose has been studied using in vitro and in vivo loops of the rat jejunum.2. At low sucrose concentrations (1 and 10 mM) glucose appeared in the serosal compartment of the in vitro preparation at a faster rate than fructose, but at high sucrose concentrations (50 and 100 mM) the rates of serosal transfer of the two sugars were similar. Glucose and fructose appeared in the mucosal compartment, with the rate of fructose appearance exceeding that of glucose, at all the sucrose concentrations studied.3. Phlorizin (5 x 10(-5)M) added to the mucosal medium of the in vitro preparation abolished the serosal transfer of glucose derived from 50 mM sucrose, and reduced that of fructose by 75%.4. In the absence of sodium ions, the in vitro preparation failed to transfer glucose and fructose derived from 50 mM sucrose, into the serosal compartment.5. Glucose was actively accumulated in the whole gut wall of the in vivo preparation to concentrations higher than those in the plasma at 50 and 100 mM, but not at 10 mM sucrose concentrations. Fructose was also actively accumulated to about half the extent of glucose, but reached tissue concentrations greater than those in the plasma, at each sucrose concentration.6. The whole wall concentrations of glucose and fructose derived from sucrose added to the lumen continued to rise when the blood supply to the in vivo preparation was terminated.7. No increase in the in vivo whole wall concentrations of glucose and fructose were detected when sucrose was added to the lumen together with concentrations of glucose sufficient to saturate the monosaccharide transport systems.8. The results favour the view that disaccharide hydrolysis and resulting hexose transfer are sequential, separate events.


Nanochannel-driven rapid capture of sub-nanogram level biomarkers for painless preeclampsia diagnosis.
Preeclampsia (PE) is a pregnancy-specific hypertensive syndrome recognized as the leading cause of maternal and fetal morbidity and mortality worldwide. Painful blood-collection procedures or low accuracy of non-invasive approaches require faster, patient-friendly, and more sensitive diagnostic technologies. Here we report a painless, highly sensitive detection platform using nanoporous microneedles (nMNs) that enables rapid capture of biomarkers present at sub-nanogram levels. The highly porous nanostructures on the nMN surface were prepared by anodization of aluminum MN and then functionalized by immobilization of capture antibodies to detect target biomarkers based on an immunoassay method. The immuno-functionalized nMN array demonstrated rapid capture of an estrogen (E2) biomarker for PE following a 1-min incubation and exhibited a concentration-dependent change in fluorescence intensity over the E2 range of 0.5 ng mL-1 to 1000 ng mL-1 after treatment with fluorescence-detection antibodies. Remarkably, the nMN patch selectively detected sub-nanogram-levels of E2 in subcutaneous interstitial fluid from rats with increased diagnostic accuracy as compared with commercial immunoassay kits. This bio-functionalized nMN platform showed improved biosensing capability for multiple PE-related biomarkers, including hormones and proteins. Furthermore, this painless method demonstrated efficacy as a point-of-need diagnostic platform using portable smartphone-based fluorescence microscope to obtain fluorescence images of biomarker-captured nMN arrays.


Valorization of solid wastes from chestnut industry processing: Extraction and optimization of polyphenols, tannins and ellagitannins and its potential for adhesives, cosmetic and pharmaceutical industry.
The aim of the current study was to evaluate the potential of chestnut peels to produce pomaces enhanced with tannins to be used in the formulations of wood adhesives, leather tanning or as natural antioxidants in food, cosmetic and pharmaceutical industry. An analytical procedure was planned as 2 factorial design to analyze the influence of solvent (water, Na2SO3 and NaOH at different concentrations of 1, 2, 4, and 8% in water) and extraction time (30, 60, and 120, 240, 480 and 960min) on extraction yield, pH, Stiasny index, and tannins. HPLC-diode array detector equipped with an ionization mass spectrophotometer was used to assess the polyphenol composition. Our results showed that both extraction properties and phytochemicals were significantly affected (P<0.001) by all independent factors. The main tannins identified were the hydrolyzable gallic acid, vescalagin castalagin and ellagic acid, and the condensed epigallocatechin, catechin and epicatechin. The solvent 1% Na2SO3 was more effective to extract the condensed tannins whilst hydrolyzable tannins were extracted efficiently by 1% NaOH. The multivariable analysis and the Pearson's correlation coefficients showed a direct association between Stiasny number and the average levels of condensed tannins.


High-resolution sedimentary records of some organochlorine pesticides in Yamzho Yumco Lake of the Tibetan Plateau: Concentration and composition.
Sediment cores from lakes have been used to reconstruct the historic deposition of persistent organic pollutants (POPs), especially in remote alpine areas. To reconstruct the deposition history of organochlorine pesticides (OCPs) in the Tibetan Plateau (TP), two sediment cores from Yamzho Yumco Lake were drilled and dated with high-resolution, from which 23 OCPs were analyzed in greater detail. Regarding several legacy compounds, concentration peaks in the cores were observed in the 1970s, corresponding to the heavy usage of these compounds around the world. In addition, another peak was found at the end of the 1990s, which was explained as the addition of OCPs released from melting glaciers or the cryosphere due to global warming. Furthermore, it was found that the transformation or degradation of OCPs after deposition in the lake was limited by comparing the values of isomeric ratios at different times, such as (DDD+DDE)/∑DDTs, o,p'-DDT/p,p'-DDT and β-HCH/∑HCHs. Such results ensure that the detected OCPs in the lake sediments really reflect their characteristics at the time of deposition. On the other hand, weak environmental self-purification of the OCPs made them last longer in the TP than in other regions. Reasonably, the fragile ecological environment of the TP would be threatened not only by legacy POPs that remain in the sediments and soils but also by POPs released from melting glaciers or the cryosphere in the next few decades under the influence of global warming. Our research provides an insight into the influence of global warming and glacial melting on the environment of the TP, and further work to gain a better understanding of the environmental processes of POPs in the TP is ongoing.


Presbyopic correction on the cornea.
The aim of this systematic review was to synthesize and appraise the evidence of the benefits of presbyopic correction on the cornea for visual function. Comprehensive search was conducted in MEDLINE using keywords like "presbylasik", "presbyopic refractive surgery", "corneal pseudoaccommodation" and "corneal multifocality". We reviewed corrected and uncorrected visual acuities for distance and near (uncorrected distance visual acuity (UDVA), uncorrected near visual acuity (UNVA), corrected distance visual acuity (CDVA), distance corrected near visual acuity (DCNVA), corrected near visual acuity (CNVA)), along with the refractive outcomes in spherical equivalent (SE) and astigmatism comparing the differences observed between preoperative myopic and hyperopic patients, as well as among techniques. Thirty-one studies met the inclusion and quality criteria. Monovision provides excellent distance and near uncorrected acuities, but with a 17% retreatment and a 5% reversal rate. Initial multifocal ablations result in 12% loss of 2 or more lines of CDVA, and a 21% retreatment rate. Laser Blended Vision provides excellent UDVA, but with a 19% retreatment rate. Initial experiences with Supracor show moderate predictability and a 22% retreatment rate. Intracor results in 9% loss of 2 or more lines of CDVA. KAMRA provides excellent UDVA, with only a 1% retreatment rate, but a 6% reversal rate. Initial experiences with PresbyMAX provided excellent UNVA and DCNVA, showing excellent predictability and a 1% reversal rate. The findings have implications for clinicians and policymakers in the health-care industry and emphasize the need for additional trials examining this important and widely performed clinical procedure.


Correction to: Assessing the integration of solar power projects: SWOT-based AHP-F-TOPSIS case study of Turkey.
The correct affiliation of the 1st Author is shown in this paper.


The Association of Pulmonary Flow Characteristics With Cardiac Function in Tricuspid Valve Malformation Fetuses: Study With Two-Dimensional Speckle Tracking Echocardiography.
To explore the relationship between cardiac function and pulmonary flow characteristics in tricuspid valve malformation (TVM) fetuses by conventional and speckle tracking echocardiography. Eighty-eight TVM fetuses were retrospectively included and compared with 80 normal controls. TVM fetuses in each trimester were divided into two subgroups according to pulmonary flow characteristics: those with normal pulmonary flow (TVM-N) and those with reduced or absent pulmonary flow (TVM-R/A). Cardiac measurements, Celermajer index, and Simpson-Andrews-Sharland (SAS) score were obtained. Speckle tracking echocardiography was used to compute ventricular deformation parameters. TVM fetuses demonstrated significantly lower global longitudinal strain (GLS) and strain rate for both ventricles than controls (all P < .05). When compared with TVM-N, left ventricular (LV) ejection fraction, LV fractional area change, right ventricular (RV) fractional area change, LVGLS, and RVGLS were significantly reduced (all P < .05), while the RV diameter Z-score, RV/LV diameter, SAS score, and Celermajer index were obviously increased in TVM-R/A in both trimesters (all P < .05). Both LVGLS and RVGLS correlated with cardiothoracic circumference ratio, RV diameter Z-score, RV/LV diameter, Celermajer index, and SAS score (all P < .01). There was a slow decline for LVGLS and RVGLS in TVM-N fetuses throughout the gestation. Conversely, these two parameters worsened rapidly in TVM-R/A group. TVM fetuses present biventricular dysfunction by deformation analysis in the second and third trimester of pregnancy. TVM fetuses with reduced or absent pulmonary flow exhibit significantly greater impairment and more rapid deterioration of cardiac function, which may contribute to poor outcomes.


Microchip electrophoresis coupled with on-line magnetic separation and chemiluminescence detection for multiplexed immunoassay.
A facile and universal strategy for multiplexed immunoassay is proposed. The strategy is based on microchip electrophoresis (MCE) coupled with on-line magnetic separation and chemiluminescence (CL) detection. The system consisted of a microchip, an electromagnet, and a photomultiplier. The realization of multiplexed immunoassay protocol involves sampling magnetic nanoparticles (MNPs) labeled antibodies, N-(4-aminobutyl)-N-ethyl-isoluminol (ABEI) labeled antigens and free antigens in the precolumn reactor, on-line immunoreaction, capturing the MNPs-immunocomplexes, and the separation of unconjugated ABEI-labeled antigens. After on-line magnetic separation, the free ABEI-labeled antigens were transported into the separation channel, and mixed with hydrogen peroxide (H(2) O(2) ) in the presence of horseradish peroxidase in the postcolumn reactor, and producing CL emission. Using this arrangement, multiple analytes could be measured simultaneously by performing the technical operations for a single assay. As a proof-of-concept, the multiplexed immunoassay was evaluated for the simultaneous determination of five model analytes (i.e. hydrocortisone, corticosterone, digoxin, testosterone, and estriol). The results exhibited excellent precision and sensitivity, the relative standard deviations for nine times detection were lower than 4.7% for all the five components, and the detection limits of five analytes were in the range of 3.6-4.9 nM. The MCE system was validated using two human serum-based control samples containing five analytes.


Fluorescent and cooperative ion pair receptor based on tolan for Na+ (or Li+) and HSO4-: logic AND gate.
A tolan derivative was synthesized as a fluorescent and cooperative ion pair receptor. As both Na+ and HSO4- ions were complexed to the receptor, only substantial fluorescence was quenched. Thus, it also acts as a logic AND gate.


Higher plant origins and the phylogeny of green algae.
5S rRNA sequences from six additional green algae lend strong molecular support for the major outlines of higher plant and green algae phylogeny that have been proposed under varying naming conventions by several authors. In particular, the molecular evidence now available unequivocally supports the existence of at least two well-separated divisions of the Chlorobionta: the Chlorophyta and the Streptophyta (i.e., charophytes) (according to the nomenclature of Bremer). The chlamydomonad 5S rRNAs are, however, sufficiently distinct from both clusters that it may ultimately prove preferable to establish a third taxon for them. In support of these conclusions 5S rRNA sequence data now exist for members of four diverse classes of chlorophytes. These sequences all exhibit considerably more phylogenetic affinity to one another than any of them show toward members of the other cluster, the Streptophyta, or the two Chlamydomonas strains. Among the Charophyceae, new 5S rRNA sequences are provided herein for three genera, Spirogyra, Klebsormidium, and Coleochaete. All of these sequences and the previously published Nitella sequence show greater resemblance among themselves and to the higher plants than they do to any of the other green algae examined to date. These results demonstrate that an appropriately named taxon that includes these green algae and the higher plants is strongly justified. The 5S rRNA data lack the resolution needed, however, to unequivocally determine which of several subdivisions of the charophytes is the sister group of the land plants. The evolutionary diversity of Chlamydomonas relative to the other green algae was recognized in earlier 5S rRNA studies but was unanticipated by ultrastructural work.(ABSTRACT TRUNCATED AT 250 WORDS)


Neonatal clinical examination and selective ultrasound screening are not reliable for the early diagnosis of hip dysplasia: A retrospective cohort study.
To compare the incidence, clinical features and timing of initial treatment of early- and later-presenting developmental dysplasia of the hip (DDH) in infancy in the context of a risk-based selective ultrasound screening programme. We conducted a retrospective observational study of infants born at a tertiary perinatal centre in Melbourne, Australia, between July 2016 and June 2020 and treated for DDH up to 18 months of age at a tertiary paediatric centre. The incidence of DDH was calculated for early- and later-presenting cases. Clinical features and ultrasound indications were analysed using multivariate logistic regression. There were 192 infants with DDH requiring treatment, 100 (52%) of which were not detected by universal neonatal hip examination and selective ultrasound screening based on risk factors (overall incidence 6.3 per 1000 livebirths). The median age at which treatment was commenced was delayed by 8 weeks for the later-presenting group compared to those detected through screening (16.7 vs. 7.9 weeks, P < 0.001). Skin crease asymmetry was associated with later presentation (adjusted odds ratio (aOR) 44.03, 95% confidence interval (CI) 9.99-333), whilst breech presentation (aOR 0.08, 95% CI 0.02-0.26) and Barlow/Ortolani test positivity (aOR 0.06, 95% CI 0.01-0.25) were associated with early presentation. Only 39 (1.45%) infants receiving a screening ultrasound had DDH. Approximately half of all infants with DDH were not detected by combining risk-based selective ultrasound screening with neonatal clinical findings, highlighting the failure of this screening method to reliably detect a significant proportion of dysplastic infant hips.


Bioassay guided isolation and identification of anti-Acanthamoeba compounds from Tunisian olive leaf extracts.
Pathogenic Acanthamoeba strains are causative agents of Granulomatous Amoebic Encephalitis (GAE) and Acanthamoeba keratitis (AK) worldwide. The existence of the cyst stage complicates Acanthamoeba therapy as it is highly resistant to antibiotics and physical agents. The aim of this study was to investigate the activity of Limouni olive leaf cultivar against the trophozoite stage of Acanthamoeba. The ethyl acetate and methanol extracts of this variety were tested against Acanthamoeba castellanii Neff. The ethyl acetate extract of olive leaf was the most active showing an IC50 of 5.11±0.71μg/ml of dry extract. Bio-guided fractionation of this extract was conducted and led to the identification of three active compounds namely oleanolic and maslinic acids and oleuropein which could be used for the development of novel therapeutic approaches against Acanthamoeba infections.


Ultrastructural and ultracytochemical differences between megakaryoblastic leukemia in children and adults. Analysis of 49 patients.
Acute megakaryoblastic leukemia (AMKL) has two peaks in distribution of incidence (in adults and children 1 to 2 years of age) and is frequently seen in children with Down syndrome. The current study was undertaken to disclose whether there were any differences between these groups. Electron microscopic and ultrastructural cytochemical features of 49 children and adults with a AMKL or chronic myelogenous leukemia (CML) in megakaryoblastic crisis were compared. Blast cells from children with AMKL, including those with and without Down syndrome, had immature features lacking typical alpha granules and a demarcation membrane system (DMS). However, blast cells from patients with AMKL with Down syndrome had more theta, electron-lucent, and basophil-like granules, suggesting that the blast cells had more potential to differentiate into other cell lines than megakaryocytes. The AMKL blast cells of adult patients showed a higher percentage of platelet peroxidase (PPO) positivity than other subgroups, and they occasionally contained typical alpha granules and DMS. This indicated that the blast cells of adults with AMKL were more mature than those of children and CML in megakaryoblastic crisis. By electron microscopic analysis, leukemic megakaryoblasts differed between children with AMKL with and without Down syndrome, adults with AMKL, and patients with CML in megakaryoblastic crisis.


[The effect of different concentrations of surface-active substances on the size of the circadian rations and on the duration of food passage in Lymnaea stagnalis infected with Echinostoma revolutum parthenitae].
In solutions of the detergent "Kristall" (0.9 and 90 mg.l.-1) there were established statistically reliable differences in the values of day rations and duration of food passing both in infected and free from infection molluscs. In noninfected individuals day rations increase 1.5 to 5 times and the duration of food passing 5 to 7 times. In infected animals the value of day rations decrease and the duration of food passing increases less intensively.


Activation of the alternative complement pathway by human B cell lymphoma lines is associated with Epstein-Barr virus transformation of the cells.
Activation of the alternative complement pathway by human B cell lymphoma lines is correlated with the presence of Epstein Barr virus (EBV) in the cell genome. EBV-negative B cell lymphoma lines produce little activation of the alternative pathway as measured either by C3 deposition on the cell surface or C3 conversion and consumption of alternative pathway activity in the supernatant serum. By contrast, EBV-positive sublines derived by in vitro EBV conversion of EBV-negative parental lines produce considerable activation of the alternative pathway. This membrane-associated complement-activating mechanism reflects an EBV-induced membrane change in these cells and may provide a mechanism whereby EBV-transformed cells are controlled in vivo.


Development of microsatellite markers for Hyacinth macaw (Anodorhynchus hyacinthinus) and their cross-amplification in other parrot species.
Hyacinth macaw Anodorhynchus hyacinthinus is the largest parrot of the world and is considered vulnerable to extinction due to its habitat loss and illegal trade associated to the international pet market demand. Genetic studies on this species are still incipient to generate a consistent characterization of the population dynamics and to develop appropriate conservation strategies. In this sense, microsatellite markers may support the detection of a population genetic structure for this bird species. However, at this time, none Hyacinth macaw species-specific primers for microsatellite loci have been so far established. This study aimed to develop and characterize polymorphic microsatellite markers for A. hyacinthinus and to check for their cross-amplification in other parrot species. Sequences containing repeated dinucleotide motifs were prospected and optimized from a genomic library that was enriched for microsatellites using magnetic beads. The analyses of 43-57 samples from wild individuals of three distinct Brazilian subpopulations led to the characterization of five polymorphic microsatellite loci. Allele richness per locus ranged from two to 12. Three loci exhibited observed heterozygosity values higher than 50%, but the overall average value among all loci was close to 45%. In addition, successful primer cross-amplification was verified in seven other investigated species of Neotropical parrots. The newly developed markers have shown to be potentially useful for in situ and ex situ population studies to support future conservation actions of Hyacinth macaw and other parrots.


Evaluation of an eye drop guide to aid self-administration by patients experienced with topical use of glaucoma medication.
Methods to improve patient compliance with prescribed topical use of glaucoma medication are sorely needed. A guide for the topical administration of ocular medication was suggested as such a tool. We investigated whether eye drop self-administration would be improved with use of the guide. An eye drop guide, a funnel-shaped device designed to fit within the contour of the orbital margins, was offered to 114 patients for use at home with their glaucoma medication. A questionnaire asking about the ease of eye drop self-administration was administered before and after 1 week's use of the device. Of the 111 patients who accepted the guide, 93 returned a usable completed questionnaire. Seventy-four percent said that they found it easier to administer their eye drops without the guide and preferred to do so (chi2 = 21.77, p < 0.01). Patients using more doses per day found it more difficult to administer their drops using the guide (r = -0.233, p < 0.05), and those who had been using drops longer preferred not to continue using the guide (r = -0.222, p < 0.05). Patients who found the written instructions clearer found it easier to administer their drops using the guide (r = 0.329, p < 0.05). Although intended to ease the administration of eye drops, the guide proved to be counterproductive. Further work is necessary to investigate other methods of improving patient compliance with prescribed topical use of ocular medications.


Analytical sensitivity of the Rapid Antigen Test kits for detection of SARS-CoV-2 Omicron variant BA.2 sublineage.
The severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) Omicron was classified as a variant of concern in November 2021. The sublineage BA.2 spreads rapidly worldwide. Currently, there is a lack of data for the parallel comparison of Rapid Antigen Test (RAT) Kits to detect SARS-CoV-2 Omicron BA.2. We evaluated the analytical sensitivity of 12 RAT kits to detect Omicron BA.2 in the present study. Analytical sensitivity was determined by means of the limit of detection (LOD). We prepared a dilution set using a respiratory specimen collected from a COVID-19 patient infected by Omicron BA.2. The reverse transcription-polymerase chain reaction was used as a reference method. The LOD results showed that all 12 RAT kits had comparable analytical sensitivity to detect Omicron BA.2. The RAT kits selected in the current study may be used for the first-line screening of the rapid spreading Omicron BA.2.


Cross-sectional associations of food insecurity with smoking cigarettes and heavy alcohol use in a population-based sample of adults.
Smoking, alcohol abuse and dual substance use represent significant public health problems and are thought to contribute to significant disparities in morbidity and mortality in the United States. Associations of food insecurity with these substances are poorly characterized, particularly concerning heavy alcohol consumption and dual substance use. Additionally, demographic factors may moderate these associations. In a cross-sectional, representative sample of Wisconsin adults (n = 1616), logistic regression was used to examine whether food insecurity was associated with cigarette smoking, heavy alcohol use and dual substance use. Interactions of food insecurity with gender, marital status and age group tested moderation by demographic factors. Food insecurity was associated with cigarette smoking in men and women (odds ratio (OR) = 3.0, 95% confidence interval (CI) = 2.1-4.4); however, only among men was food insecurity associated with heavy alcohol use (OR = 1.5; 95% CI = 1.0-2.2) and dual substance use (OR = 5.2; 95% CI = 1.5-18.6). Marital status and age group did not moderate associations of food insecurity with substance use. Food insecurity and substance use are highly correlated. Heavy alcohol consumption and dual use were positively correlated with food insecurity among men, but not among women. Longitudinal analyses are needed to disentangle temporality and underlying pathways of observed associations. Additionally, future work should determine whether interventions and policies targeting food insecurity can simultaneously reduce cigarette and alcohol use.


Ferroptosis as a mechanism of oligodendrocyte loss and demyelination in experimental autoimmune encephalomyelitis.
Ferroptosis, distinct from necrosis, autophagy and apoptosis, is a unique form of regulated cell death,and is a potential pathogenic mechanism of neuronal loss and defunction in many neurodegenerative disorders. Recent studies have shown a presence of iron deposition in the central nervous system (CNS) of patients with multiple sclerosis (MS). However, whether ferroptosis is involved in the pathogenesis of MS remains unclear. In the present study, we tested certain classical biomarkers of ferroptosis in the spinal cord of mice with experimental autoimmune encephalomyelitis (EAE), an animal model of MS, to substantiate the relationship between ferroptosis and oligodendrocyte (OL) loss and demyelination. Our results revealed decreased levels of key molecules in glutathione antioxidant mechanisms, including system xC (xCT) and glutathione peroxidase 4 (GPX4) in spinal cord of EAE mice, with evident lipid peroxidation in OLs. Moreover, transferrin receptor and ferritinophagy further catalyzed the generation of lipid reactive oxygen species through the fenton reaction, which induced OL death and demyelination at disease peak of EAE. This phenomenon was largely reversed by administering Fer-1, an inhibitor of ferritin phagocytosis, further validating the key role of ferritin phagocytosis in EAE. Taken together, these findings demonstrate that OL loss and demyelination may be induced in EAE through, at least in part, a mechanism of ferroptosis.


Human cytomegalovirus infection in a retinoblastoma cell line in vitro.
The focus of these studies was to determine whether the Y79 human retinoblastoma cell line could function as a good in vitro model system for studying human cytomegalovirus (HCMV) infection. Y79 cells were exposed to an HCMV mutant carrying a LacZ gene, and the resulting beta-galactosidase expression in infected cells was assessed by flow cytometry. The extent to which the three classes of viral gene products immediate early, early, and late proteins - were expressed was analyzed by immunohistochemical staining and Western blotting. Infected Y79 cells were also co-cultivated on human foreskin fibroblast (SF cell) cultures to recover virus. Infection of Y79 cells with the virus resulted in beta-galactosidase expression as detected by flow-cytometric analysis. Immunohistochemical staining revealed that a portion of Y79 cells expressed antigens reactive to monoclonal antibodies against immediate early, early, and late HCMV proteins. The 43-kDa early gene product was also detected by Western blotting. Infected Y79 cells co-cultivated on SF cell cultures yielded infectious foci, which turned blue following X-gal staining, demonstrating productive HCMV infection in the Y79 cells. These results demonstrate that while HCMV can productively infect Y79 cultures, it does so in a highly inefficient manner, leading these authors to conclude that this cell line does not provide a particularly good model system to study HCMV infection.


Ibrutinib Is Associated With Increased Cardiovascular Events and Major Bleeding in Older CLL Patients.
Early ibrutinib trials showed an association between ibrutinib use and risk of bleeding and atrial fibrillation (AF) in younger chronic lymphocytic leukemia (CLL) patients. Little is known about these adverse events in older CLL patients and whether increased AF rates are associated with increased stroke risk. To compare the incidence of stroke, AF, myocardial infarction, and bleeding in CLL patients treated with ibrutinib with those who were treated without ibrutinib in a linked SEER-Medicare database. The incidence rate of each adverse event for treated and untreated patients was calculated. Among those treated, inverse probability weighted Cox proportional hazards regression models were used to calculate HRs and 95% CIs for the association between ibrutinib treatment and each adverse event. Among 4,958 CLL patients, 50% were treated without ibrutinib and 6% received ibrutinib. The median age at first treatment was 77 (IQR: 73-83) years. Compared with those treated without ibrutinib, those treated with ibrutinib had a 1.91-fold increased risk of stroke (95% CI: 1.06-3.45), 3.65-fold increased risk of AF (95% CI: 2.42-5.49), a 4.92-fold increased risk of bleeding (95% CI: 3.46-7.01) and a 7.49-fold increased risk of major bleeding (95% CI: 4.32-12.99). In patients a decade older than those in the initial clinical trials, treatment with ibrutinib was associated with an increased risk of stroke, AF, and bleeding. The risk of major bleeding is higher than previously reported and underscores the importance of surveillance registries to identify new safety signals.


Calibrating Population Stratification in Association Analysis.
In genetic association studies, it is necessary to correct for population structure to avoid inference bias. During the past decade, prevailing corrections often only involved adjustments of global ancestry differences between sampled individuals. Nevertheless, population structure may vary across local genomic regions due to the variability of local ancestries associated with natural selection, migration, or random genetic drift. Adjusting for global ancestry alone may be inadequate when local population structure is an important confounding factor. In contrast, adjusting for local ancestry can more effectively prevent false positives due to local population structure. To more accurately locate disease genes, we recommend adjusting for local ancestries by interrogating local structure. In practice, locus-specific ancestries are usually unknown and must be inferred. For recently admixed populations with known reference ancestral populations, locus-specific ancestries can be inferred accurately using some hidden Markov model-based methods. However, SNP-wise ancestries cannot be accurately inferred when ancestral population information is not available. For such scenarios, we propose employing local principal components (PCs) to present local ancestries and adjusting for local PCs when testing for gene-phenotype association.


Doxycycline improves cage activity, but not exercised, supraspinatus tendon and muscle in a rat model.
The objective of this study was to investigate the effects of doxycycline, a broad-spectrum MMP inhibitor, on cage activity and exercised supraspinatus tendon and muscle using a Sprague-Dawley rat model of non-injurious exercise. Because exercise may alter muscle and tendon MMP activity and matrix turnover, we hypothesized that doxycycline would abolish the beneficial adaptations found with exercise but have no effect on cage activity muscle and tendon properties. Rats were divided into acute or chronic exercise (EX) or cage activity (CA) groups, and half of the rats received doxycycline orally. Animals in acute EX groups were euthanized 24 h after a single bout of exercise (10 m/min, 1 h) on a flat treadmill. Animals in chronic EX groups walked on a flat treadmill and were euthanized at 2 or 8 week time points. Assays included supraspinatus tendon mechanics and histology and muscle fiber morphologic and type analysis. Doxycycline improved tendon mechanical properties and collagen organization in chronic cage activity groups, which was not consistently evident in exercised groups. Combined with exercise, doxycycline decreased average muscle fiber cross-sectional area. Results of this study suggest that administration of doxycycline at pharmaceutical doses induces beneficial supraspinatus tendon adaptations without negatively affecting the muscle in cage activity animals, supporting the use of doxycycline to combat degenerative processes associated with underuse; however, when combined with exercise, doxycycline does not consistently produce the same beneficial adaptations in rat supraspinatus tendons and reduces muscle fiber cross-sectional area, suggesting that doxycycline is not advantageous when combined with activity.


Patient perceptions of empathy in primary care telephone consultations: A mixed methods study.
Clinical empathy can enhance patient outcomes. This study examined patients' perceptions of empathy in primary care consultations delivered by telephone. A mixed methods study was nested in a larger feasibility study conducted May-October 2020. Adults reporting a UK primary care consultation in the previous 2 weeks completed an online survey. A sample of survey respondents participated in a semi-structured qualitative interview. Interviews were analysed thematically. Survey respondents (n = 359) rated practitioners as between 'good' and 'very good' at established patient-reported indicators of clinical empathy. Telephone consultations were rated slightly lower than face-to-face or other consultations. 30 survey respondents were interviewed. Three qualitative themes identified how telephone consultations can shape clinical empathy: setting for an empathic encounter; feeling connected; being acknowledged. Primary care patients typically perceive good levels of clinical empathy in telephone consultations; specific features of telephone consultations may facilitate and/or hinder clinical empathy. To ensure patients feel listened to, acknowledged and understood, practitioners may need to increase their empathic verbalisations in telephone consultations. By using verbal responses to demonstrate active listening and by clearly describing and/or implementing next steps in management, practitioners may be able to enhance clinical empathy in telephone consultations.


Evaluation of subacute toxicity of methanolic/aqueous preparation of aerial parts of O. sanctum in Wistar rats: Clinical, haematological, biochemical and histopathological studies.
Ocimum sanctum, commonly known as Holy Basil or Tulsi has been used in Ayurveda as a demulcent, stimulant, expectorant; in the treatment of bronchitis, skin infections, malaria, diarrhoea, dysentery, arthritis, gastric and inflammatory disorders. We have previously shown that methanolic/aqueous extract of O. sanctum did not induce genotoxicity and other toxic effects in acute oral toxicity study. In the present report, we have performed sub-acute toxicity of methanolic/aqueous preparation of O. sanctum in Wistar rats to evaluate whether it induced any chronic toxic effects. In subacute toxicity study, animals received O. sanctum extract (OSE) by oral gavage at the doses of 250, 500 and 1000 mg/kg/day (n=5/group/sex) for 28 days. At the end of the study, the animals were sacrificed and evaluated for the effect of OSE on clinical, haematological, biochemical and histopathological parameters. The rats treated with OSE did not show any change in body weight, food and water consumption, motor activity, sensory reactivity and foot splay measurements. There were no significant changes in haematological, pathological and biochemical parameters; and histopathology of tissues (liver, kidney, spleen, heart, and testis/ovary) among rats of either sex. OSE at a dose of 1000 mg/kg showed significant increase of Mean corpuscular hemoglobin (MCH) (19.8 ± 0.8; 18.7 ± 0.5) and mean corpuscular hemoglobin concentration (MCHC) (41.8 ± 1.1; 39.3 ± 0.7) in male and female rats in comparison to their respective controls (MCH: 17.7 ± 0.3; 17.4 ± 0.3; MCHC: 37.8 ± 0.5; 36.1 ± 0.2). Urine parameters (appearance, blood, nitrate, leucocyte, glucose, ketone, pH, protein and specific gravity) in both the male and female rats were comparable to their respective controls. In addition, no changes were observed in the vital organs of rats at macroscopic and microscopic levels. Our results showed that oral administration of OSE was not toxic to male and female Wistar rats upto the highest dose tested, thereby suggesting its clinical usefulness.


Percutaneous intervention for unprotected left main disease prior to explantation of a left ventricular assist device.
Percutaneous coronary intervention of unprotected left main coronary arterial disease is an alternative to surgical revascularization in selected patients. In this report, a patient with an implanted left ventricular assist device (LVAD) underwent successful coronary intervention prior to its planned removal. The implanted LVAD clearly assisted the technical performance of the intervention.


Development of a prediction protocol for the screening of metabolic associated fatty liver disease in children with overweight or obesity.
The early detection and management of children with metabolic associated fatty liver disease (MAFLD) is challenging. To develop a non-invasive and accurate prediction protocol for the identification of MAFLD among children with overweight/obesity candidates to confirmatory diagnosis. A total of 115 children aged 8-12 years with overweight/obesity, recruited at a primary care, were enrolled in this cross-sectional study. The external validation was performed using a cohort of children with overweight/obesity (N = 46) aged 8.5-14.0 years. MAFLD (≥5.5% hepatic fat) was diagnosed by magnetic resonance imaging (MRI). Fasting blood biochemical parameters were measured, and 25 candidates' single nucleotide polymorphisms (SNPs) were determined. Variables potentially associated with the presence of MAFLD were included in a multivariate logistic regression. Children with MAFLD (36%) showed higher plasma triglycerides (TG), insulin, homeostasis model assessment of insulin resistance (HOMA-IR), alanine aminotransferase (ALT), aspartate transaminase (AST), glutamyl-transferase (GGT) and ferritin (p < 0.05). The distribution of the risk-alleles of PPARGrs13081389, PPARGrs1801282, HFErs1800562 and PNLPLA3rs4823173 was significantly different between children with and without MAFLD (p < 0.05). Three biochemical- and/or SNPs-based predictive models were developed, showing strong discriminatory capacity (AUC-ROC: 0.708-0.888) but limited diagnostic performance (sensitivity 67%-82% and specificity 63%-69%). A prediction protocol with elevated sensitivity (72%) and specificity (84%) based on two consecutive steps was developed. The external validation showed similar results: sensitivity of 70% and specificity of 85%. The HEPAKID prediction protocol is an accurate, easy to implant, minimally invasive and low economic cost tool useful for the early identification and management of paediatric MAFLD in primary care.


Effect of different doses of BCG on an allogeneic cell-mediated antitumor response in mice.
The effect of BCG on the development of cell-mediated cytotoxicity (CMC) in the spleens of allogeneic tumor-bearing (C57BL/6 X A)F1 mice was studied. BCG-treated tumor-bearing animals elicited a stronger CMC response than did the untreated tumor-bearing animals. Furthermore, the degree of activation was dependent on different doses of BCG, so that if the BCG was administered at doses above or below the optimum dose, a less marked stimulation of the CMC resulted. The combination of BCG given before the tumor implant plus an intratumor inoculation resulted in a significantly higher CMC than did either treatment alone.


The Role of Crystalline Intermediates in Mechanochemical Cyclorhodation Reactions Elucidated by in-Situ X-ray Powder Diffraction and Computation.
The occurrence of crystalline intermediates in mechanochemical reactions might be more widespread than previously assumed. For example, a recent study involving the acetate-assisted C-H activation of N-Heterocycles with [Cp*RhCl2 ]2 by ball milling revealed the formation of transient cocrystals between the reagents prior to the C-H activation step. However, such crystalline intermediates were only observed through stepwise intervallic ex-situ analysis, and their exact role in the C-H activation process remained unclear. In this study, we monitored the formation of discrete, stoichiometric cocrystals between benzo[h]quinoline and [Cp*RhCl2 ]2 by ball milling using in-situ synchrotron X-ray powder diffraction. This continuous analysis revealed an initial cocrystal that transformed into a second crystalline form. Computational studies showed that differences in noncovalent interactions made the [Cp*RhCl2 ]2 unit in the later-appearing cocrystal more reactive towards NaOAc. This demonstrated the advantage of cocrystal formation before the acetate-assisted metalation-deprotonation step, and how the net cooperative action of weak interactions between the reagents in mechanochemical experiments can lead to stable supramolecular assemblies, which can enhance substrate activation under ball-milling conditions. This could explain the superiority of some mechanochemical reactions, such as acetate-assisted C-H activation, compared to their solution-based counterparts.


Quality consistency evaluation on three species of North Patrininae herba by high-performance liquid chromatography coupled with electrospray ion trap time-of-flight mass spectrometry and network pharmacology approaches.
North Patrininae herba, a perennial herbaceous plant, has long been used in traditional Chinese medicine to treat appendicitis, enteritis, and dysentery. Sonchus arvensis L., Sonchus oleraceus L., and Ixeris chinensis (Thunb.) Nakai are used as substitutes for North Patrininae in different regions, but the consistency of chemical composition and efficacy of these three species is still unknown. In this study, a detailed chemical analysis was carried out of the extract obtained from Sonchus arvensis L., Ixeris chinensis (Thunb.) Nakai and Sonchus oleraceus L. and a chemical component not previously reported in Ixeris chinensis (Thunb.) Nakai was found-Luteolin-7-O-(6''-malonylglucoside). The mechanism of action of the extract against inflammation and type II diabetes was investigated using network pharmacology and analysis of blood-absorbed components following oral dosing of rats. Finally, a highly accurate and reliable method was established for quality control purposes. The results showed that Sonchus arvensis L. and Sonchus oleraceus L. may be considered potential resources of a medicinal compound, whereas Ixeris chinensis (Thunb.) Nakai requires further evaluation.


Lipid composition of cultured B16 melanoma cell variants with different lung-colonizing potential.
Lipid components influence several cell surface properties that are critical in different stages of the metastatic process. In this study, we examined whether the different lung-colonizing potential of B16-F1 and B16-F10 melanoma cells could be related to a characteristic lipid profile. The lipid analyses, carried out on the same cell cultures used for the assay of lung-colonizing potential, revealed characteristics in the lipid composition of both B16-F1 and B16-F10 melanoma cells that are common to other systems of malignant cells: a high level of 18:1 associated with low proportions of polyunsaturated fatty acids in phospholipids, accumulation of ether-linked lipids and absence of complex gangliosides. The two B16 melanoma variants differed significantly only with respect to ether-linked lipids, due to a higher level of alkyl-PC in B16-F10 than in B16-F1.


Differential cytokine expression and regulation of human anti-pig xenogeneic responses by modified porcine dendritic cells.
Porcine dendritic cells (DC) are likely to be pivotal cells in the initiation of stimulatory and potential tolerogenic responses to xenoantigens, however, there are limited studies characterizing these antigen presenting cells. Porcine PBMC (CD172a(+)) were cultured with GM-CSF and IL-4 and phenotype and functional capabilities assessed. Lipopolysaccharide (LPS), IL-10, and IL-3 were added to the GM-CSF/IL-4 DC cultures to determine phenotypic and functional changes. Quantitative real-time polymerase chain reaction (PCR) for key cytokines was performed and the modified porcine DC were further assessed by primary mixed lymphocyte reaction to determine the effect of LPS, IL-10, and IL-3 on stimulatory capability. Porcine PBMC (CD172(+)) cultured with GM-CSF and IL-4 produced cells with DC morphology, which were major histocompatability complex (MHC) class II(+), CD14(-/lo), and CD1a(lo). Addition of IL-10 or IL-3 to GM-CSF/IL-4 DC cultures produced cells with lower levels of MHC class II and higher levels of antigen uptake consistent with less mature DC. Quantitative real-time PCR of DC showed the addition of IL-10 induced an increase in IL-10 mRNA, no detectable IL-12, and reduced IL-6 mRNA. The addition of IL-3 to DC cultures decreased IL-12, IL-6 and tumor necrosis factor (TNF), with no change in IL-10 mRNA. GM-CSF/IL-4 DC induced strong human lymphocyte proliferation, compared with significantly reduced stimulatory capacity induced by IL-10 and IL-3 treated DC cultures. The profound effect on differential DC cytokine profile and reduced human anti-pig responses has important therapeutic implications in xenotransplantation. The mechanism of altered regulation warrants further investigation.


Collecting high-quality pancreatic tissue for experimental study from organ donors with signs of β-cell autoimmunity.
The aim of this study was to create a new research strategy to obtain high-quality pancreatic tissues from subjects with preclinical or clinical type 1 diabetes, which would open up new avenues for studying the mechanisms of the β-cell damaging process in humans. A nationwide collaboration network (the PanFin network) was established in Finland to start an on-call screening of diabetes-associated autoantibodies from deceased organ donors and subsequent processing of pancreases from autoantibody-positive donors. This protocol was integrated into the national organ transplantation procedure. Only a few modifications were needed to the normal transplantation practices. One additional blood sample was obtained from donors for autoantibody analyses, the transplantation team was informed about the autoantibody result and the pancreas of autoantibody-positive donors was transported to the core laboratory. Altogether, 307 donors were screened and 22 (7.2%) were positive for at least one autoantibody and 3 tested positive for two or more autoantibodies out of the five tested (islet cell antibodies, insulin autoantibodies and autoantibodies to glutamic acid decarboxylase, islet antigen 2 and zinc transporter 8). The quality of collected pancreatic tissue was superior to that from autopsies and allowed the detection of both RNA and proteins. The study protocol was proven feasible to be carried out on a nationwide scale. It did not interfere with the normal transplantation activities and provided valuable tissue material for research.


Chromosomal localization and genomic organization of the human retinal rod Na-Ca+K exchanger.
The retinal rod Na-Ca+K exchanger is a unique calcium extrusion protein found only in the outer segments of retinal rod photoreceptors. Rod Na-Ca+K exchanger cDNA (NCKX1) has been cloned from bovine and human retinas. Here, we have used fluorescent in situ hybridization and radiation hybrid mapping to localize the human NCKX1 gene to chromosome 15q22. We have determined the genomic organization of human rod NCKX1 and found one intron in the 5' untranslated region and eight introns within the coding region.


Synaptic mechanisms underlying onset and progression of memory deficits caused by hippocampal and midbrain synucleinopathy.
Cognitive deficits, including working memory, and visuospatial deficits are common and debilitating in Parkinson's disease. α-synucleinopathy in the hippocampus and cortex is considered as the major risk factor. However, little is known about the progression and specific synaptic mechanisms underlying the memory deficits induced by α-synucleinopathy. Here, we tested the hypothesis that pathologic α-Synuclein (α-Syn), initiated in different brain regions, leads to distinct onset and progression of the pathology. We report that overexpression of human α-Syn in the murine mesencephalon leads to late onset memory impairment and sensorimotor deficits accompanied by reduced dopamine D1 expression in the hippocampus. In contrast, human α-Syn overexpression in the hippocampus leads to early memory impairment, altered synaptic transmission and plasticity, and decreased expression of GluA1 AMPA-type glutamate receptors. These findings identify the synaptic mechanisms leading to memory impairment induced by hippocampal α-synucleinopathy and provide functional evidence of the major neuronal networks involved in disease progression.


PrP amyloid plaques in Creutzfeldt-Jakob disease of short duration: immunohistochemical studies of 5 cases from Poland.
We report here PrP-immunohistochemistry performed on brains from CJD cases from Poland. Only one of five definitive CJD cases exhibited typical PrP-immunoreactive kuru-like plaques and this was case of a short duration. We thus confirm the low frequency of PrP plaques in CJD of Eastern and Central European origin.


The transcribed genome and the heritable basis of essential hypertension.
Gene expression can be now assessed quantitatively and comprehensively. In addition to reflecting the specialized differentiation of the cell or tissue type from which gene expression is sampled, it also manifests patterns determined by inheritance. Thus gene expression is a phenotypic trait, at least when assessed comprehensively. This trait shows familial aggregation and segregation patterns indicative of an inherited contribution. The molecular evolution of genes includes mutations affecting regulatory sequences in the genome that influence gene expression in cis and in trans. Such mutations may increase in frequency in a population either by genetic drift or by selection. Traits of gene expression, acting alone or in concert with other gene expression traits, may generate phenotypes that extend beyond transcript abundance. Indeed, the divergence of species and the traits that distinguish related species appear to rely importantly on inherited divergence in the control of gene expression. Variation in gene expression may contribute to the pathogenesis of a prevalent human disease trait that shows heritability--essential hypertension. Along with other common heritable diseases, hypertension susceptibility arises from the actions of multiple genome sequence variations. The identity of such variation has proven elusive when sought by methods that have been successfully applied to Mendelian diseases. This review explores the potential to uncover hypertension genes by exploiting quantitative variation in the heritable control of gene expression.


Simultaneous resection of colorectal primary tumour and synchronous liver metastases.
The surgical strategy for treatment of synchronous colorectal liver metastases remains controversial. The outcome and overall survival of patients presenting with such metastases, treated either by simultaneous resection or by delayed resection, were evaluated. From 1987 to 2000, 97 patients presented with synchronous colorectal liver metastases, of whom 35 (36 per cent) underwent a simultaneous resection and 62 patients (64 per cent) a delayed resection. Simultaneous resection was considered prospectively for patients with fewer than four unilobar metastases. Age, blood transfusion requirements, operating time, duration of inflow occlusion, hospital stay and mortality rate were similar in the two groups. The morbidity rate did not differ significantly (23 per cent after simultaneous resection and 32 per cent after delayed resection). The location of the primary tumour and extent of liver resection did not influence the morbidity rate significantly in the simultaneous resection group. The overall survival rate was 94, 45 and 21 per cent at 1, 3 and 5 years respectively after simultaneous resection, and 92, 45 and 22 per cent after delayed resection. In selected patients, simultaneous resection of the colorectal primary tumour and liver metastases does not increase mortality or morbidity rates compared with delayed resection, even if a left colectomy and/or a major hepatectomy are required.


Staged approach to combined repair of pectus excavatum and lesions of the heart.
The combined repair of severe pectus excavatum and cardiac lesions remains technically difficult. We report our experience with a patient undergoing such a combined repair of pectus and implantation of an aortic valved conduit in the setting of the Marfan syndrome. The procedure was staged such that the dissection and bony mobilization was done at the first procedure before heparin administration in combination with a sternal split and subsequent aortic repair. The second stage of sternal repositioning and cartilage resection was completed several days later with excellent cardiopulmonary and cosmetic results. The technique and a review of the literature are included.


Design and Synthesis of Biotinylated Bivalent Carboline Derivatives as Potent Antitumor Agents.
Compound 6, a novel β-carboline comprising two 1-methyl-9H-β-carboline-3-carboxylic acids and a biotin moiety conjugated together using tris(2-aminoethyl)amine, was synthesized and tested for its cytotoxicity toward MCF-7 and HepG2 cell lines and antitumor potency in an S180 tumor-bearing mouse model. Compound 6 was delivered via biotin receptor-mediated endocytosis and exerted its therapeutic effects by intercalation binding with DNA. In vivo antitumor evaluations of 6 revealed that it is efficacious and exhibits low systemic toxicity.


Modulation of extracellular glutamate concentration in rat brain slices by cystine-glutamate exchange.
1. The role of cystine-glutamate exchange in controlling the extracellular glutamate concentration in the central nervous system was examined by whole-cell clamping neurons in rat brain slices, and using their glutamate receptors as sensors of extracellular glutamate concentration. 2. Applying cystine to cerebellar slices generated a membrane current in Purkinje cells which was abolished by glutamate receptor blockers. Similar cystine-evoked currents were seen in pyramidal cells of frontal cortex slices. 3. Control experiments on non-N-methyl-D-aspartate (non-NMDA) receptors in enzymatically isolated Purkinje cells showed that cystine did not produce a current in slice Purkinje cells by directly activating glutamate receptors, nor by potentiating the action of background levels of glutamate on receptors. Experiments on isolated salamander Muller cells showed that cystine did not block Na+-dependent GLAST glutamate transporters (homologous to the transporters in the Bergmann glia ensheathing the Purkinje cells), nor did it block the current produced by EAAT4 and EAAC1 glutamate transporters in Purkinje cells. Thus the cystine-evoked current in Purkinje cells is not due to a rise in extracellular glutamate concentration caused by block of Na+-dependent uptake. 4. The dependence of cystine-evoked current on cystine concentration in slice Purkinje cells could be fitted by a Michaelis-Menten relation with a Km of 250 microM. The Km predicted from this for cystine activating glutamate efflux is less than 140 microM, because of the non-linear dependence on glutamate concentration of the Purkinje cell current. The current evoked by 1 mM cystine was little affected by removal of extracellular chloride or addition of 1 mM furosemide (frusemide), but was potentiated by 1 mM 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid (DIDS). 5. These data suggest that external cystine generates a current in slice Purkinje cells by activating cystine-glutamate exchange in cells of the slice, releasing glutamate which activates non-NMDA receptors in the Purkinje cell membrane.


The effectiveness of inpatient rehabilitation in the acute postoperative phase of care after transtibial or transfemoral amputation: study of an integrated health care delivery system.
To compare outcomes between lower-extremity amputees who receive and do not receive acute postoperative inpatient rehabilitation within a large integrated health care delivery system. An observational study using multivariable propensity score risk adjustment to reduce treatment selection bias. Data compiled from 9 administrative databases from Veterans Affairs Medical Centers. A national cohort of veterans (N=2673) who underwent transtibial or transfemoral amputation between October 1, 2002, and September 30, 2004. Not applicable. One-year cumulative survival, home discharge from the hospital, and prosthetic limb procurement within the first postoperative year. After reducing selection bias, patients who received acute postoperative inpatient rehabilitation compared to those with no evidence of inpatient rehabilitation had an increased likelihood of 1-year survival (odds ratio [OR]=1.51; 95% confidence interval [CI], 1.26-1.80) and home discharge (OR=2.58; 95% CI, 2.17-3.06). Prosthetic limb procurement did not differ significantly between groups. The receipt of rehabilitation in the acute postoperative inpatient period was associated with a greater likelihood of 1-year survival and home discharge from the hospital. Results support early postoperative inpatient rehabilitation following amputation.


Usefulness of the ice-cream cone pattern in computed tomography for prediction of angiomyolipoma in patients with a small renal mass.
A morphologic contour method for assessing an exophytic renal mass as benign versus malignant on the basis of the shape of the interface with the renal parenchyma was recently developed. We investigated the usefulness of this morphologic contour method for predicting angiomyolipoma (AML) in patients who underwent partial nephrectomy for small renal masses (SRMs). From January 2004 to March 2013, among 197 patients who underwent partial nephrectomy for suspicious renal cell carcinoma (RCC), the medical records of 153 patients with tumors (AML or RCC) ≤3 cm in diameter were retrospectively reviewed. Patient characteristics including age, gender, type of surgery, size and location of tumor, pathologic results, and specific findings of the imaging study ("ice-cream cone" shape) were compared between the AML and RCC groups. AML was diagnosed in 18 patients and RCC was diagnosed in 135 patients. Gender (p=0.001), tumor size (p=0.032), and presence of the ice-cream cone shape (p=0.001) showed statistically significant differences between the AML group and the RCC group. In the multivariate logistic regression analysis, female gender (odds ratio [OR], 5.20; 95% confidence interval [CI], 1.45 to 18.57; p=0.011), tumor size (OR, 0.34; 95% CI, 0.12 to 0.92; p=0.034), and presence of the ice-cream cone shape (OR, 18.12; 95% CI, 4.97 to 66.06; p=0.001) were predictors of AML. This study confirmed a high incidence of AML in females. Also, the ice-cream cone shape and small tumor size were significant predictors of AML in SRMs. These finding could be beneficial for counseling patients with SRMs.


Excessive gestational weight gain and the risk of gestational diabetes: Comparison of Intergrowth-21st standards, IOM recommendations and a local reference.
To compare the abilities of Intergrowth-21st standards, Institute of Medicine (IOM) recommendations and a Chinese reference on gestational weight gain (GWG) to identify women at risk of gestational diabetes (GDM) and GDM-related adverse outcomes. A retrospective cohort study was conducted on 13,366 women delivering live singleton infants between 2013 and 2017 in Tongzhou district of Beijing, China. Poisson regression with robust error estimates was used to estimate risk ratios (RRs) of GDM in different GWG groups according to three standards. There were 39.97%, 46.31% and 30.03% of women gaining weight above Intergrowth-21st standards, IOM recommendations and the Chinese reference respectively. Women with GWG above Intergrowth-21st standards and the Chinese reference had 27% (aRR, 1.27 95% CI, 1.18-1.37) and 30% (aRR, 1.30; 95% CI, 1.21-1.40) increased risks of GDM respectively, as compared to 22% (aRR, 1.22; 95% CI, 1.13-1.32) for IOM recommendations. GWG above either of these three standards was associated with macrosomia and cesarean delivery (P < 0.05). Compared with IOM recommendations, GWG above Intergrowth-21st standards or the Chinese reference was associated with higher risks of GDM and GDM-related adverse outcomes. Furthermore, these two prospective standards could additionally assess the severity of abnormal GWG and are feasible for dynamic monitoring.


Age dependent trans-cellular propagation of human tau aggregates in Drosophila disease models.
Tauopathies is a class of neurodegenerative disorders which involves the transformation of physiological tau into pathogenic tau. One of the prime causes reported to drive this conversion is tau hyperphosphorylation and the subsequent propagation of pathogenic protein aggregates across the nervous system. Although past attempts have been made to deduce the details of tau propagation, yet not much is known about its mechanism. A better understanding of this aspect of disease pathology can prove to be beneficial for the development of diagnostic and therapeutic approaches. For the first time, we demonstrate that the human tau possesses an intrinsic property to spread trans-cellularly in the fly nervous system irrespective of the tau allele or the neuronal tissue type. Aggregate migration restricted by targeted down-regulation of a specific kinase, elucidates the role of hyper-phosphorylation in its movement. On the contrary to the previous models, our study delivers an easy and rapid in-vivo model for comprehensive examination of tau migration pathology. Henceforth, the developed model would not only be immensely helpful in uncovering the mechanistic in-depths of tau propagation pathology but also aid in modifier and/or drug screening for amelioration of tauopathies.


Holt-Oram syndrome associated with the hypoplastic left heart syndrome.
We present the first case of Holt-Oram syndrome associated with the lethal congenital heart defect of hypoplastic left heart syndrome. The possible pathophysiological link is explored and the need for careful genetic and cardiologic evaluation in these patients is reiterated.


Lack of a requirement for a maternal humoral immune response to establish or maintain successful allogeneic pregnancy.
In general, breeding pairs are not major histocompatibility complex (MHC)-compatible, and therefore the fetoplacental unit can be considered as a natural allograft. In many mammals pregnancy leads to the production of nonlytic antibodies of antipaternal MHC specificity. It has been suggested that these protect the semiallogeneic fetus from rejection by acting as blocking or enhancing factors--or, alternatively, that they are part of a humoral response involved in the establishment of normal pregnancy. These hypotheses were tested in allomated mice made B cell deficient by continuous treatment with alpha IgM mu antiserum. The status of the maternal immune system was assessed by in vivo antibody production, in vitro mitogen responses, and allograft rejection. By these criteria B cell function could not be demonstrated in alpha IgM mu treated female mice, but T cell responses were unaffected. Allogeneic pregnancy, however, was not compromised by this humoral immune system dysfunction--litter size and neonatal survival being the same in the alpha IgM mu and control serum-treated groups. These results indicate that a maternal humoral immune response is not essential for the establishment of pregnancy or the survival of the semiallogeneic fetus.


Henri Marie Raymond de Toulouse-Lautrec-Montfa (1864-1901): artistic genius and medical curiosity.
Toulouse-Lautrec was born in Albi in South West France. His parents were first cousins; this consanguineous marriage was responsible for his ugly deformities. Lautrec's life in Montmartre became the setting for much of his art and for his deliberate slow self-destruction. In addition to his artistic ability he was courageous, a natural leader, an accomplished chef, a connoisseur of fine wine, intelligent, humorous and very generous.


The "Athlete's Heart": relation to gender and race.
Long-term athletic training is associated with changes in cardiac morphology, commonly described as "athlete's heart." Although numerous studies have investigated the effects of training on cardiac dimensions, most are limited to male Caucasian athletes, and few data are available regarding the effect of long-term exercise training on the woman's heart. This article reviews the athlete's heart in relation to gender and race.


Epidemic cycles and environmental pressure in colonial Quebec.
Research on historical populations in Europe finds that infectious disease epidemics appear to induce predictable cycles in age-specific mortality. We know little, however, about whether such cycles also occurred in less dense founder populations of North America. We used high-quality data on the Quebecois population from 1680 to 1798 to examine the extent to which age-specific mortality showed predictable epidemic cycles. We further examined whether environmental pressures-temperature, lack of precipitation, or crop failure-may have set the stage for the emergence of epidemics. We applied autoregressive, integrated, moving average time series methods to annual counts of period mortality for the following age groups: < 1 year, 1 to < 5 years, 5 to < 15 years, 15 to < 50 years, and 50 years and above. These methods controlled for other patterns (e.g., trend) before empirically identifying cycles. Results indicate a strong seven-year cycle in mortality among infants and children under seven years of age. Warm temperatures (across Quebec overall) and relatively dry years (in Eastern Quebec) also predicted an increased risk of mortality in infancy and childhood, although these environmental variables appear to act independently of the epidemic cycle pattern. Findings indicate a strong seven-year epidemic cycle in historical Quebec which afflicted naïve birth cohorts not previously exposed to the prior epidemic. We contend that smallpox epidemics likely contributed to this cycle. The seven-year cycle occurred only in the latter half of the test period (post 1740) with increasing size of the colony and population concentration in urban areas.


Lymphoid tissue reactions in rheumatoid arthritis.
Frequently, the immune cell infiltrate of chronically inflamed tissues develops functional germinal centres and acquires structural features of secondary lymphoid organs. Ectopic lymphoid structures occur in peripheral tissues not only during autoimmune diseases but also in tumors (reactive infiltrate), chronic infections and graft rejection, indicating a strong link between lymphoid neogenesis and persistent antigen driven immune/inflammatory responses. There has been a renewed interest in ectopic lymphoid neogenesis, as better understanding of the mechanisms underpinning this process could contribute to elucidate the bio-pathological mechanisms involved in transition from acute-self resolving to chronic immunological aggression as well as identify novel therapeutic targets. Here we critically review recent clinical and biological studies addressing the role of ectopic lymphoid neogenesis specifically in rheumatoid arthritis.


Radiation-induced degradation of methyl orange in aqueous solutions.
Degradation of methyl orange under gamma-irradiation was investigated. The reactions followed pseudo first-order kinetics. Apparent degradation rate constant, estimated through linear regression analysis, increased with an increase of dose rate and a decrease of initial methyl orange concentration. Degradation of methyl orange was significantly accelerated under oxidative condition, but was slightly enhanced under reductive condition. However, the result of decoloration was better under reductive condition than oxidative one. An analysis on the intermediates using Fourier transform infrared and gas chromatography/mass spectrometry demonstrates that the radiolytic degradation of methyl orange was processed with different C-N cleavages under oxidative and reductive conditions.


Identification and characterisation of a small marker chromosome using non-isotopic in situ hybridisation with X and Y specific probes.
A 13 year old male with mild mental retardation, obesity, and poor secondary sexual differentiation was found to have a 46,X,+,mar karyotype. In situ hybridisation with X and Y specific probes proved the marker to be composed of Y centromeric and short arm material.


Vascular endothelial growth factor gene and the response to anti-vascular endothelial growth factor treatment for choroidal neovascularization in high myopia.
To investigate the association between the vascular endothelial growth factor (VEGF) gene polymorphism and the response to anti-VEGF treatment for choroidal neovascularization (CNV) in highly myopic eyes. Retrospective cohort study. A total of 357 unrelated highly myopic Japanese patients with axial lengths ≥26.0 mm in both eyes were eligible, and 83 patients who received anti-VEGF therapy for CNV and could be followed for more than 1 year were included. We genotyped a functional single nucleotide polymorphism in the VEGF gene, rs2010963. The associations between the distribution of the rs2010963 genotype and the number of eyes with maintained or improved visual acuity (VA) were analyzed. Furthermore, multivariable logistic regression analysis was performed to adjust for 7 possible prognostic factors, including age, sex, CNV size, CNV location, administration of loading dose, pretreatment VA, and number of additional treatments. The primary end point was maintenance of VA, and secondary end points were progression of chorioretinal atrophy (CRA) and recurrence of CNV. Mean age and mean axial length were not significantly different among 3 genotypes of rs2010963. The percentage of eyes with maintained or improved VA was significantly higher with the G allele of rs2010963 (P =0.016), and stepwise analysis revealed that both rs2010963 and CNV size were associated with VA maintenance (P =0.040 and 0.033, respectively). The secondary analysis revealed that administration of a loading dose was significantly associated with both CRA progression (P =0.031) and recurrence of CNV (P =0.020), whereas rs2010963 was not. These results suggest that the VEGF polymorphism influences the VA prognosis in highly myopic eyes with CNV within 1 year after anti-VEGF treatment. This association was still observed after removing its confounding effect through CNV size. The rs2010963 polymorphism was not associated with CNV recurrence or CRA progression, which indicates that these changes are not tied to intrinsic factors and may be controllable by improving treatment methods.


Molecular identification of Haemosporidia in avian endemics of Gorgona Island within a context for the eastern tropical Pacific region.
Island bird populations and their obligate blood parasites are of interest for understanding the accumulation of biodiversity and the evolutionary relationship with their mainland congeners. We examined avian Haemosporidia cytochrome b gene among terrestrial birds on Gorgona Island National Park, Colombia. Three Haemoproteus haplotype groups found on Gorgona Island have a higher genetic similarity to Haemoproteus found in the eastern tropical Pacific than those documented in Africa, Asia, Europe and Oceania. Two of the haplotype groups on the island are generalists in terms of infecting multiple hosts and their wide geographical distribution within the eastern tropical Pacific region, a third Haemoproteus haplogroup appears endemic to Gorgona Island. The overall prevalence of haemosporidian parasites is 57,9% for birds on Gorgona island, which is higher than local reports of prevalence documented in other archipelagos or the mainland. The island population of Cyanerpes cyaneus gigas seems to be the most susceptible to Haemoproteus infection when compared to Thamnophilus atrinucha gorgonae and Coereba flaveola gorgonae. Our findings support an ubiquitous pattern of endemism among organisms including avian haemosporidian parasites on Gorgona Island and also highlight the potential exposure of island bird populations to avian malaria.


Capsular polysaccharide from a fish-gut bacterium induces/promotes apoptosis of colon cancer cells in vitro through Caspases' pathway activation.
Among the widespread malignancies colorectal cancer is the most lethal. Treatments of this malignant tumor include surgery for lesions and metastases, radiotherapy, immunotherapy, and chemotherapy. Nevertheless, novel therapies to reduce morbidity and mortality are demanding. Natural products, such as polysaccharides, can be a valuable alternative to sometimes very toxic chemotherapeutical agents, also because they are biocompatible and biodegradable biomaterials. Microbial polysaccharides have been demonstrated to fulfill this requirement. In this paper, the results about the structure and the activity of a capsular polysaccharide isolated from the psychrotroph Pseudoalteromonas nigrifaciens Sq02-Rifr, newly isolated from the fish intestine, have been described. The characterization has been obtained by spectroscopic and chemical methods, and it is supported by the bioinformatic analysis. The polymer activates Caspases 3 and 9 on colon cancer cells CaCo-2 and HCT-116, indicating a promising antitumor effect, and suggesting a potential capacity of CPS to induce apoptosis.


[Medical treatment of pulmonary hypertension: what's new?].
Pulmonary hypertension (PH) is a chronic progressive disease of the pulmonary circulation of multifactorial causes. The current diagnostic classification of PH distinguishes five main groups, which have as a common feature an increased pulmonary arterial pressure and pulmonary resistance. The classification differentiates pulmonary arterial hypertension (PAH), PH due to left heart disease, PH in lung diseases and/or hypoxia, chronic thromboembolic pulmonary hypertension (CTEPH), and PH with unclear/multifactorial mechanisms. Recent advances in basic research with the approval of new drugs and the establishment of therapeutic strategies, mainly in PAH and CTEPH, require a differentiated view of the disease, a careful diagnosis and initiation of therapy, and regular follow-ups. In this article, we provide an overview of the complex drug therapy currently available for PAH patients.


The ratio of serum testosterone-to-prostate specific antigen predicts prostate cancer in hypogonadal men.
We determined whether the ratio of serum testosterone to prostate specific antigen might provide diagnostic value regarding the risk of prostate cancer in a population of hypogonadal men undergoing prostate biopsy. The study population consisted of 184 consecutive men with symptomatic hypogonadism and prostate specific antigen 4.0 ng/ml or less who underwent prostate biopsy before the anticipated initiation of testosterone therapy. All men had testosterone 300 ng/dl or less. Testosterone concentrations were converted to ng/ml, eg 270 ng/dl equals 2.7 ng/ml, to calculate the testosterone-to-prostate specific antigen ratio. Mean patient age was 58.5 years. There were 154 men with benign biopsies and 30 with cancer. Testosterone concentrations were similar in the prostate cancer and noncancer groups, although mean prostate specific antigen was higher in the prostate cancer group. The testosterone-to-prostate specific antigen ratio was inversely related to prostate cancer risk (OR 0.49, 95% CI 0.33-0.74). On multivariate analysis performed by logistic regression neither age nor prostate specific antigen was be predictive of prostate cancer. However, the testosterone-to-prostate specific antigen ratio remained strongly associated with prostate cancer risk. An ROC for the testosterone-to-prostate specific antigen ratio suggested that a ratio of below 1.8 was diagnostic for prostate cancer, while values below this threshold were associated with an OR of 3.17 (95% CI 1.17-8.59) for prostate cancer. A low ratio of testosterone to prostate specific antigen is an independent predictor of prostate cancer in hypogonadal men with prostate specific antigen 4.0 ng/ml or less. Ratios less than 1.8 were associated with a greater than 3-fold increase in prostate cancer risk.


Association between arterial stiffness and the presence of cerebral small vessel disease markers.
We investigated the effect of arterial stiffness on the severity of enlarged perivascular spaces (EPVSs) and cerebral microbleeds (CMBs) at different brain locations. A total of 854 stroke patients underwent both brachial-ankle pulse wave velocity (baPWV) measurement and brain MRI. The extent of EPVS was separately rated at the levels of the basal ganglia (BG) and centrum semiovale (CS). The CMBs were categorized as strictly lobar CMB and deep CMB. The patients were categorized according to baPWV quartiles, and multivariable logistic regressions were performed to evaluate whether the baPWV increment was independently associated with each cerebral SVD marker at different locations. The odds ratio (OR) with 95% confidence interval (CI) was derived on the reference of the first quartile. Severe EPVSs at BG and CS were detected in 243 (28.5%) and 353 patients (41.3%), respectively. The increment of baPWV quartiles was associated with both severe BG EPVS burden (Q4: OR = 2.58, CI = 1.45-4.60) and severe CS EPVS burden (Q4: OR = 2.06, CI = 1.24-3.42). Deep CMBs were found in 259 patients (30.3%), and strictly lobar CMBs were found in 170 patients (19.9%). Multivariable logistic regression model revealed deep CMB was independently associated with the baPWV increment (Q4: OR = 2.52, CI = 1.62-3.94). However, strictly lobar CMB had a neutral relationship with baPWV. Increased arterial stiffness is consistently associated with the presence of deep CMB and severe EPVS burden at the BG and CS, suggesting a common pathophysiologic mechanism.


Black-white differences in maternal age, maternal birth cohort, and period effects on infant mortality in the US (1983-2002).
We investigate three interrelated sources of change in infant mortality rates over a 20year period using the National Center for Health Statistics (NCHS) linked birth and infant death cohort files. The effects of maternal age, maternal birth cohort, and time period of childbirth on infant mortality are estimated using a modified age/period/cohort (APC) model that identifies age, period, cohort effects. We document black-white differences in the patterning of these effects and find that maternal age effects follow the predictable U-shaped pattern, net of period and cohort, but with a less steep gradient in the black population. The largest relative maternal age-specific disparity in IMR occurs among older African American mothers. Cohort effects, while considerably smaller than age and period effects, present an interesting pattern of a modest decline in IMR among later cohorts of African American mothers coupled with an increasing IMR among the same cohorts of non-Hispanic whites. However, period effects dominate the time trends, implying that period-related technologies overwhelmingly shape US infant survival in today's population. These general findings are mirrored in APC analyses carried out for several leading underlying causes of infant mortality.


Medium optimization for enhanced production of recombinant lignin peroxidase in Pichia pastoris.
Different cultivation conditions and parameters were evaluated to improve the production and secretion of a recombinant Phanerochaete chrysosporium lipH8 gene in Komagataella phaffii (Pichia pastoris). The recombinant lipH8 gene with its native secretion signal was successfully cloned and expressed in Komagataella phaffii (Pichia pastoris) under the control of the alcohol oxidase 1 promoter (PAOX1). The results revealed that co-feeding with sorbitol and methanol increased rLiP secretion by 5.9-fold compared to the control conditions. The addition of 1 mM FeSO4 increased LiP activity a further 6.0-fold during the induction phase. Moreover, the combination of several optimal conditions and parameters yielded an extracellular rLiP activity of 20.05 U l-1, which is more than ten-fold higher relative to standard growth conditions (BMM10 medium, pH 6 and 30 °C). Extracellular activity of a recombinant LiP expressed in P. pastoris increased more than ten-fold when co-feeding sorbitol and methanol as carbon sources, together with urea as nitrogen source, FeSO4 supplementation, lower pH and lower cultivation temperature.


Red cell esterase-d-polymorphism in the Veneto population.
630 individuals and 102 mother/child pairs from the Veneto provinces were examined for their red cell EsD by Cellogel electrophoresis. Compared with the horizontal starch-gel electrophoresis no differences in the zymogram patterns have been found. The gene frequencies observed (EsD1 = 0.8556; EsD2 = 0.1444) differ only slightly from those reorted for various European populations and are nearly identical with data for other Italian populations. Family studies confirmed the Mendelian co-dominant inheritance.


Integrated imaging of non-small cell lung cancer recurrence: CT and PET-CT findings, possible pitfalls and risk of recurrence criteria.
To review the role of imaging in the diagnosis of recurrent disease in previously treated non-small cell lung cancer (NSCLC) and discuss the imaging pitfalls. A comprehensive review of published literature on CT and PET imaging of NSCLC recurrence was performed. Diagnostic and prognostic values are discussed. Representative imaging examples are illustrated. Up to 30% of NSCLC recurrences present as loco-regional, involving treated hemithorax and ipsilateral lymph nodes, while 70% present as metachronous distant metastases. CT and PET-CT play an important role in the early detection of recurrence; indications for imaging vary depending on pathological features. Imaging plays a central role in the identification of recurrence and may predict prognosis. Lung cancer recurs after surgery in 30% to 75% of patients. CT and PET-CT are crucial in identification of loco-regional recurrence. Knowledge of potential pitfalls is essential, especially for parenchymal or nodal recurrence. CT can diagnose metastases but further examinations (PET-CT, MRI) are often needed. Morphological and functional imaging criteria may help in predicting recurrence.


Effect of an organic acid blend in Nile tilapia growth performance, immunity, gut microbiota, and resistance to challenge against francisellosis.
Organic acids (OAs) are a class of feed additives that have prophylactic and inhibitory properties against pathogenic bacteria. In this study, we investigated growth performance, innate immune response, gut microbiota, and disease resistance against Francisella orientalis F1 in Nile tilapia (Oreochromis niloticus) fed different doses of Bacti-nil®Aqua, a blend of short- and medium-chain OAs. For 21 days, 680 juvenile tilapias were fed a control diet or diets supplemented with a 0.3% (D3) or 0.5% (D5) OA blend. The feed conversion rate of fish fed the 0.5% enriched diet was considerably lower (p < 0.05) than that of the fish fed the basal diet. Lysozyme and serum bactericidal activities were significantly elevated following OA administration. After infection, no differences in the diversity and composition of gut microbiota were observed between the groups. After the bacterial challenge, the mortality was significantly lower in group D5 (p < 0.01). The diet supplemented with Bacti-nil®Aqua (Adisseo) improved the immune response and resistance of tilapia juveniles against F. orientalis infection. Thus, this OA blend could serve as a feed additive with good activity against F. orientalis.


Chinese herbal medicine for irinotecan-induced diarrhea: A systematic review and meta-analysis.
Irinotecan is a novel first-line therapy for colorectal cancer, but the toxicity and side effects include diarrhea without satisfactory treatments. Chinese herbal decoction (CHD) is an effective complementary and alternative prevention and therapy for irinotecan induced diarrhea (IID). This systematic review and meta-analysis of randomized controlled trials (RCTs) aims to assess the preventive effect of CHD in the treatment of IID. Seven databases (PubMed, COCHRANE, EMBASE, CNKI, VIP, Wanfang, and CBM) were screened for random controlled trials on the prevention and treatment of IID by CHD from January 1980 to May 2022. The Cochrane Collaboration Risk of Bias (ROB 2.0) was applied for bias risk assessment, and the Grading Recommendation Assessment, Development, and Evaluation (GRADE) for quality of evidence. Meta-analysis was conducted with RevMan 5.3 software. In addition, a subgroup analysis was conducted on different grades of diarrhea, the incidence and duration of diarrhea, the selection of specific Chinese herbal medicine decoction, and the incidence of adverse reactions. Risk ratios (RR) and 95% confidence intervals (CIs) were calculated for all data by combining the meta-analysis with fixed or random-effects models based on outcome heterogeneity. Fourteen RCTs involving 1056 participants were included. The study results displayed that the incidence of IID was lower with the use of CHD than the no-treatment group (RR = 0.55, 95% CI = 0.40-0.75, P = 0.0002). CHD in combination with western medicine (WM) was more effective than WM alone for IID (RR = 0.44, 95% CI 0.23-0.84, P = 0.01). This protective effect was more pronounced for severe grade III-V diarrhea (RR = 0.41, 95% CI = 0.26-0.64, P < 0.0001). In the specific Chinese herbal medicine decoction, the Banxia Xie Xin decoction presented better effectiveness (RR = 0.18, 95% CI: 0.05-0.63, P = 0.007) than WM alone. The Huangqin decoction was the most widely studied interventional scheme (n = 5). The relative risk (RR) of the Huangqin decoction was 0.56. No obvious adverse reactions were observed. This study demonstrated that CHD has a preventive effect on IID and could be used as a complementary therapy with few side effects. However, additional large-sample, high-quality, randomized, double-blind trials are needed to guide the clinical practice scientifically. www.crd.york.ac.uk/prospero (NO: CRD42020189506).


Paul Klee's gastrointestinal maladies: A series of historical and clinical vignettes, part IV.
Paul Klee (1879-1940), the 20th century Swiss-German artist, suffered and died from complications of systemic sclerosis (SSc, scleroderma). In a series of clinical and historical vignettes the various symptoms and complications of Klee's scleroderma are described. Here, I present evidence of Klee's multiple gastrointestinal (GI) symptoms and the significant impact they had on the quality of his life.