Fifteen-year clinical prognosis and cardiovascular or limb event associated with homocysteine levels in peripheral arterial disease.
There are limited reports on the relationship between plasma homocysteine (Hcy) levels and long-term all-cause death (ACD), cardiovascular events, or limb events in patients with peripheral arterial disease (PAD). We examined the relationship between plasma Hcy levels and 15-year these events in PAD patients. We performed a prospective cohort study in 955 PAD patients. The patients were divided into four groups based on plasma Hcy levels with median (interquartile range). The endpoints were cumulative incidences of ACD, major adverse cardiovascular events (MACE), and MACE plus limb events (MACLE). The incidences of ACD, MACE, and MACLE were correlated with plasma Hcy levels (P < 0.05). In multiple regression analysis, plasma Hcy had positive correlations with C-reactive protein (CRP), men, and critical limb ischemia (CLI) and negative correlations with estimated glomerular filtration rate (eGFR) and high-density lipoprotein cholesterol (p < 0.05). In Cox multivariate analysis, higher Hcy (HR 1.614, 95 % CI 1.229-2.119, p = 0.001), age, CRP, brain natriuretic peptide (BNP), D-dimer, lower body mass index, ankle brachial pressure index (ABI), serum albumin, eGFR, CLI, coronary heart disease (CHD), cerebrovascular disease, and diabetes were related to ACD; higher Hcy (HR 1.242, 95 % CI 1.004-1.535, p = 0.045), age, BNP, lower ABI, serum albumin, diabetes, and CHD were related to MACE; and higher Hcy (HR 1.290, 95 % CI 1.057-1.574, p = 0.012), BNP, lower ABI, serum albumin, CHD, and diabetes were related to MACLE (P < 0.05). Statins improved ACD, MACE, and MACLE (p < 0.01). Plasma Hcy was a risk factor for 15-year ACD, MACE, and MACLE in patients with PAD.


Thermal damage of the specimen during breast biopsy with the use of the Breast Lesion Excision System: does it affect diagnosis?
Stereotactic vacuum assisted breast biopsy (VABB) procedures remain of key importance in the diagnostic evaluation of suspicious non-palpable mammographic lesions. The Breast Lesion Excision System(®) (BLES) is an image-guided percutaneous biopsy method that utilizes radiofrequency (RF) in order to retrieve an intact-tissue biopsy specimen. The purpose of this study is to determine the effects of RF application on the tissue specimen and the possible interference with the histopathologic results. In this study we included 265 patients with suspicious non-palpable mammographic lesions who underwent 273 stereotactic VABB procedures using the BLES. All biopsies were performed by the same surgical-radiology team under local anesthesia. The specimen thermal damage was classified according to the pathology report in 3 categories: Severe (extensive thermal damage or inability to diagnose), medium (ability to diagnose but either circumferential damage >1.5 mm or diffuse areas of thermal damage) and mild (circumferential thermal damage <1.5 mm but >0.5 mm). Radiofrequency-associated thermal damage of the specimen was observed in 14 cases (5.13 %), and was classified as severe in 5, medium in 5, and mild in 4 specimens. Within the group of RF damaged specimens, we found a significant (p < 0.05) positive correlation between fat cell content and classification of thermal damage. Although thermal damage is of concern during BLES breast biopsy, the incidence is low, and the outcome of the histopathologic assessment is not affected even in severely damaged specimens. Increased thermal damage seems to correlate with higher fat cell content of the specimen.


A bulge binding agent with novel wedge-shape topology for stimulation of DNA triplet repeat strand slippage synthesis.
Expansion of DNA repeat sequences is associated with many human genetic diseases. Bulged DNA structures have been implicated as intermediates in DNA slippage within the DNA repeat regions. Herein a bulge binding agent with novel wedge-shape topology of the aromatic moiety was designed and synthesized. The compound-bulge DNA interactions were characterized via UV melting experiments, circular dichroism and were quantitated by surface plasmon resonance with K(d) of 41.5 microM. This compound showed remarkable stimulation for DNA triplet repeat strand slippage synthesis in vitro.


Development of the pre-school child: the validation of a psychomotor screen, and the influence of the home environment on psychomotor development.
The present study was designed to validate and standardize a short psychomotor screening test on Australian four-year-old-preschool children and to assess their home environment using Caldwell's HOME Inventory. The Adelaide Psychomotor Screen (APS) is a short, 10-15 minute screening test which uses 13 separate items to assess "General" development, "Gross Motor" development, "Social/Emotional Behaviour" and "Speech". In the area of "General" development, 12 children screened as "abnormal" and 54 children screened as "normal" were further assessed by a psychologist on the McCarthy Scales of Children's Abilities. There was a high correlation between the APS "General" scores and the McCarthy (General Cognitive Index) scores (r = 0.75, p less than 0.001 for the younger children, and r = 0,90, p less than 0.001 for the older children). Caldwell's HOME Inventory takes an hour to complete, and involves a visit by the assessor to each home. The correlation between the HOME total scores and the McCarthy (General Cognitive Index) scores was r = 0.06, p less than 0.001. It is suggested that the HOME Inventory may be more valuable as a predictor of a child's future development than an index of his present developmental status. It is suggested that nurses and teachers used the APS as a screening test of the individual child, and use the HOME inventory as an assessment of the home environment.


Neuropsychologic assessment of visual disorders.
This article introduces the reader to a sample of visual tests used in the neuropsychologic assessment of patients who present with various visual deficits. As discussed, patients often present with visual abnormalities that cannot be assessed exclusively during the opthtalmologic examination, partly because these problems extend beyond the fundamental aspects of visual processing. Visual problems occur in the context of focal or diffuse brain damage. Neuropsychologic evaluation can provide valuable diagnostic information and information regarding functional strengths and weaknesses. Many visual tests have been developed for clinical use. Some of these tests have been validated with lesion analytic or neuroimaging studies, which highlight the areas of the brain presumed necessary for task performance. Knowledge regarding the neural substrates of test performance allows the clinician to identify the neuropathologic correlates of test failure, which, in turn, is relevant to differential diagnosis. A profile of functional strengths and weaknesses emerges contributing to the treatment of the patient with a visual disorder. In this article, the authors present a subset of visual tests used primarily in the clinical setting. Some of these tests measure lower-level visual deficits (e.g., Judgment of Line Orientation) and others measure higher-level visual/cognitive deficits (e.g., ROCF). Although no firm delineation of test subtypes exists, the authors divide the tests into general categories of visuoperceptual, visuospatial, visuoconstructive, and visual attention/memory. Ultimately, it is incumbent on a trained neuropsychologist to select appropriate visual tests based on the patient's described symptoms and the referral question.


Insights on adsorption of carbamazepine onto iron oxide modified diatomaceous earth: Kinetics, isotherms, thermodynamics, and mechanisms.
To ameliorate adsorbent recovery by an external magnetic field, naturally occurring diatomaceous earth (DE) was modified with iron-oxide, characterized and applied for adsorption of carbamazepine (CBZ) from synthetic wastewater using batch equilibration method. The fabricated adsorbent was characterized using XRF, XRD, SEM-EDX, FT-IR, BET surface area analysis, VSM and pH of point of zero charge (pHpzc) determination. The adsorption rate was described by the pseudo-first-order (PFO) model suggesting a physisorption controlled rate-determining step. Equilibrium adsorption data were fitted to linear and nonlinear isotherm models, viz Langmuir and Freundlich models, and were best described by Freundlich nonlinear equations implying heterogeneous multilayer adsorption. The best-fitting kinetic and isotherm model was determined using four mathematical error functions. The thermodynamic parameters, namely enthalpy (ΔH = -26.4 kJ mol-1), Gibbs free energy (ΔG = -2.22 kJ mol-1 at 298 K), entropy (ΔS = -34.0 kJ mol-1), indicated that the adsorption was a spontaneous, exothermic, and physical process. The adsorption mechanism is postulated to involve cation-π interactions. Modified diatomaceous earth is a potentially excellent, low-cost, and novel sorbent for CBZ adsorption with 88% removal in 180 min and provides a possible alternative adsorbent for wastewater treatment.


COVID-19 surveillance in Democratic Republic of Congo, Nigeria, Senegal and Uganda: strengths, weaknesses and key Lessons.
As part of efforts to rapidly identify and care for individuals with COVID-19, trace and quarantine contacts, and monitor disease trends over time, most African countries implemented interventions to strengthen their existing disease surveillance systems. This research describes the strengths, weaknesses and lessons learnt from the COVID-19 surveillance strategies implemented in four African countries to inform the enhancement of surveillance systems for future epidemics on the continent. The four countries namely the Democratic Republic of Congo (DRC), Nigeria, Senegal, and Uganda, were selected based on their variability in COVID-19 response and representation of Francophone and Anglophone countries. A mixed-methods observational study was conducted including desk review and key informant interviews, to document best practices, gaps, and innovations in surveillance at the national, sub-national, health facilities, and community levels, and these learnings were synthesized across the countries. Surveillance approaches across countries included - case investigation, contact tracing, community-based, laboratory-based sentinel, serological, telephone hotlines, and genomic sequencing surveillance. As the COVID-19 pandemic progressed, the health systems moved from aggressive testing and contact tracing to detect virus and triage individual contacts into quarantine and confirmed cases, isolation and clinical care. Surveillance, including case definitions, changed from contact tracing of all contacts of confirmed cases to only symptomatic contacts and travelers. All countries reported inadequate staffing, staff capacity gaps and lack of full integration of data sources. All four countries under study improved data management and surveillance capacity by training health workers and increasing resources for laboratories, but the disease burden was under-detected. Decentralizing surveillance to enable swifter implementation of targeted public health measures at the subnational level was a challenge. There were also gaps in genomic and postmortem surveillance including community level sero-prevalence studies, as well as digital technologies to provide more timely and accurate surveillance data. All the four countries demonstrated a prompt public health surveillance response and adopted similar approaches to surveillance with some adaptations as the pandemic progresses. There is need for investments to enhance surveillance approaches and systems including decentralizing surveillance to the subnational and community levels, strengthening capabilities for genomic surveillance and use of digital technologies, among others. Investing in health worker capacity, ensuring data quality and availability and improving ability to transmit surveillance data between and across multiple levels of the health care system is also critical. Countries need to take immediate action in strengthening their surveillance systems to better prepare for the next major disease outbreak and pandemic.


Environmental exposure to benzene, micronucleus formation and polymorphisms in DNA-repair genes: a pilot study.
This report is part of a biomarker study conducted in an Italian population with exposure to environmental benzene ranging from 1.43 to 31.41 μg/m³ (values from personal sampling). DNA damage induced by benzene is the crucial mechanism of its genotoxicity, which leads to chronic benzene poisoning, haematotoxicity and leukaemia. Therefore, genetic variation in DNA-repair genes may modulate susceptibility to benzene-induced DNA damage. In light of this, the effects of polymorphisms in DNA-repair genes (APEX1, hOGG1, NBS1, XPD, XRCC1, and XRCC3) on micronucleus (MN) formation as a biomarker of early biological effects were evaluated. A significantly higher median MN frequency was recorded in traffic wardens than in controls. However, none of the analysed polymorphisms was significantly associated with the median MN frequency. A gene-gender interaction was observed for the APEX1 genotype. The APEX1 variant genotype was associated with significantly lower median MN frequency in men, not in women. Statistical analysis did not reveal any association between the score of the protective alleles - hypothetically pushing the pathway towards optimal DNA-damage repair - and MN. Even though there are some limitations in the study, our results indicate that the general population may be exposed to benzene concentrations higher than the threshold level for air-quality standards in the European Union of 10 μg/m³. Furthermore, urban traffic wardens are exposed to significantly higher levels of benzene than individuals spending most of the time indoors. This higher exposure may contribute to DNA damage, suggesting that benzene might be implicated both as an environmental and occupational risk factor in leukaemia and other haematological diseases. In conclusion, this study suggest the need for (i) regular monitoring of traffic wardens for possible exposure to benzene, as a precautionary step to reduce the associated health risks, and (ii) more comprehensive studies in order to better elucidate the involvement of APEX1 genotypes in benzene genotoxicity.


MINSTED nanoscopy enters the Ångström localization range.
Super-resolution techniques have achieved localization precisions in the nanometer regime. Here we report all-optical, room temperature localization of fluorophores with precision in the Ångström range. We built on the concept of MINSTED nanoscopy where precision is increased by encircling the fluorophore with the low-intensity central region of a stimulated emission depletion (STED) donut beam while constantly increasing the absolute donut power. By blue-shifting the STED beam and separating fluorophores by on/off switching, individual fluorophores bound to a DNA strand are localized with σ = 4.7 Å, corresponding to a fraction of the fluorophore size, with only 2,000 detected photons. MINSTED fluorescence nanoscopy with single-digit nanometer resolution is exemplified by imaging nuclear pore complexes and the distribution of nuclear lamin in mammalian cells labeled by transient DNA hybridization. Because our experiments yield a localization precision σ = 2.3 Å, estimated for 10,000 detected photons, we anticipate that MINSTED will open up new areas of application in the study of macromolecular complexes in cells.


Clinical course of Crohn's disease in Italy.
The clinical course of Crohn's disease in 131 patients was studied for a mean period of 4.2 +/- 3.2 years. The clinical activity of the disease, expressed as percentage of patients per year in an active phase, is high in the first year (70.2 percent) and progressively decreases during subsequent years (25 percent after seven years). The percentage of patients who needed steroid treatment is high during the first year (68 percent) and falls to 19 percent after seven years. An operative risk rate of 54 percent was registered, with a probability of reoperation equal to 34 percent. Clinical relapse after the first surgery occurred in 70 percent of cases. The registered mortality was 6.9 percent, with a ratio of 6 to 1 between observed and expected mortality. In conclusion, the disease, while showing a tendency to reduce its activity over the years, is burdened by a risk of surgery and mortality which progressively increases with time.


Allele-specific non-CG DNA methylation marks domains of active chromatin in female mouse brain.
DNA methylation at gene promoters in a CG context is associated with transcriptional repression, including at genes silenced on the inactive X chromosome in females. Non-CG methylation (mCH) is a distinct feature of the neuronal epigenome that is differentially distributed between males and females on the X chromosome. However, little is known about differences in mCH on the active (Xa) and inactive (Xi) X chromosomes because stochastic X-chromosome inactivation (XCI) confounds allele-specific epigenomic profiling. We used whole-genome bisulfite sequencing in a mouse model with nonrandom XCI to examine allele-specific DNA methylation in frontal cortex. Xi was largely devoid of mCH, whereas Xa contained abundant mCH similar to the male X chromosome and the autosomes. In contrast to the repressive association of DNA methylation at CG dinucleotides (mCG), mCH accumulates on Xi in domains with transcriptional activity, including the bodies of most genes that escape XCI and at the X-inactivation center, validating this epigenetic mark as a signature of transcriptional activity. Escape genes showing CH hypermethylation were the only genes with CG-hypomethylated promoters on Xi, a well-known mark of active transcription. Finally, we found extensive allele-specific mCH and mCG at autosomal imprinted regions, some with a negative correlation between methylation in the two contexts, further supporting their distinct functions. Our findings show that neuronal mCH functions independently of mCG and is a highly dynamic epigenomic correlate of allele-specific gene regulation.


Oligonucleotide typing is a perfect tool to identify antigens stimulatory in the mixed lymphocyte culture.
An important criterion for the selection of donors for bone marrow transplantation is the grade of matching for HLA between donor and recipient. For patients that lack an HLA-identical sibling, an extending pool of unrelated volunteers for bone marrow donation is available. From these donors the best matched candidate can be selected by serological typing, followed by a mixed lymphocyte culture (MLC). Oligonucleotide genotyping for HLA class II antigens is considered to be valuable for the prediction of MLC reactivity. We investigated whether this typing method, in combination with serological typing, would cover the recognition of all MLC stimulatory determinants. One hundred thirty-six combinations of HLA-A, -B, and -DR serologically identical individuals were tested in the MLC. Additional typing for HLA-DRB and HLA-DPB by oligonucleotide genotyping made it possible to evaluate the influence of these genes on MLC reactivity. Combinations that were matched for HLA-DRB gave significantly lower responses than those that were mismatched. Nevertheless, in the matched combinations responses were observed to 94% relative response index. These responses could all be attributed to HLA-DP, since all combinations that were identical by HLA-DPB genotyping were negative in the MLC. In conclusion, with the combined use of serology and oligonucleotide genotyping, responder-stimulator combinations can be selected that are identical for all MLC stimulatory determinants.


New Advances in Supportive Care: Chemoprotective Agents as Novel Opportunities in Geriatric Oncology.
To explore the effectiveness of trilaciclib and ALRN-6924 in the prevention of cancer chemotherapy-induced toxicity in older patients. New chemoprotective agents are necessary because age is the main risk factor for chemotherapy complications that account largely for the poorer outcome of cancer in the elderly. Trilaciclib and ALRN-6924 cause a reversible block of the proliferation of normal cells through cell cycle arrest (CCA). With this mechanism, they may prevent the toxicity of cycle-active cancer treatment including neutropenia, anemia, thrombocytopenia, lymphopenia, mucositis, and alopecia. Myelopoietic growth factors may prevent neutropenia in the aged, but they may cause severe bone pain, may aggravate thrombocytopenia and anemia, and may cause myelodysplasia and acute leukemia as a late complication. The prevention of thrombocytopenia, anemia, mucositis, and alopecia is unsatisfactory at present. These complications may jeopardize the treatment outcome as they require a reduction of treatment dose/intensity and because many patients find the resulting symptoms intolerable. In three studies of patients with extensive disease small cell lung cancer (ES-SCLC), trilaciclib reduced the severity and duration of neutropenia and thrombocytopenia as well as the need for blood transfusions. In addition, it produced a significant expansion of T-cell clones. Trilaciclib received FDA approval for the prevention of chemotherapy-induced myelosuppression in patients with ES-SCLC. ALRN-6924 is currently studied in phase II study of ES-SCLC. In a phase IB of 38 patients, ALRN-6924 prevented myelosuppression to an extent comparable with trilaciclib. Both drugs proved as effective in patients 65 and older as they were in the younger ones. In an "ex vivo" study, ALRN-6924 protected the epithelial stem cells of hair follicles from taxanes and promised to prevent alopecia. The possibility that CCA of tumor cells may reduce the effectiveness of cycle-active chemotherapy is a major concern. For this reason, the use of trilaciclib, an inhibitor of CDK 4/6, should be limited to tumors with inactivated RB1, and the use of ALRN-6924, an inhibitor of P53, should be limited to tumors with inactivated P53. Chemotherapy-related toxicities limit dose intensity and contribute to significant morbidity and mortality in elderly cancer patients. Trilaciclib and ALRN-6924 are of particular interest to geriatric oncologists because of their novel mechanism of action. Ameliorating chemotherapy-induced toxicities holds the promise of transforming the practice of geriatric oncology by enabling chemotherapeutic regimens that are currently not feasible for this patient population. Specifically, these agents may prevent chemotherapy-induced neutropenia and thrombocytopenia, perhaps the most life-threatening complications of cytotoxic chemotherapy, thereby obviating the need for the use of rescue strategies such as hematopoietic growth factors. In addition, these agents offer the potential for broad tissue protection from other chemotherapy-related toxicities, including mucositis, diarrhea, and alopecia, which historically have been poorly managed. Importantly, by preventing a spectrum of chemotherapy-related toxicities, these agents may permit the administration of chemotherapy at full-dose intensity, prevent functional decline, and grant maintenance of resilience to older cancer patients. As a result, the successful prevention of chemotherapy-induced side effects may not only mitigate the costs of care but also improve patient outcomes and quality of life. Finally, chemoprotective strategies offer the opportunity to apply geriatric principles to clinical trials of cancer treatment. In particular, they may allow the testing of prolongation of "active life expectancy" as a major goal of clinical trials in elderly patients. They may also enable novel and more practical forms of clinical trials. By assessing the risk of chemotherapy-related toxicity with the Chemotherapy Risk Assessment Scale for High Age Patients (CRASH) or the Cancer and Aging Research Group (CARG) instruments, these agents may permit researchers to utilize patients as their own controls and endorse the approval of supportive care drugs based upon the risk profile of individual patients.


Current advances in the detection and removal of organic arsenic by metal-organic frameworks.
Arsenic (As) is a highly toxic heavy metal and has been widely concerned for its hazardous environmental impact. Aromatic organic arsenic (AOCs) has been frequently used as an animal supplement to enhance feed utilization and prevent dysentery. The majority of organic arsenic could be discharged from the body and evolve as highly toxic inorganic arsenic that is hazardous to the environment and human health via biological conversion, photodegradation, and photo-oxidation. Current environmental issues necessitate the development and application of multifunctional porous materials in environmental remediation. Compared to the conventional adsorbent, such as activated carbon and zeolite, metal-organic frameworks (MOFs) exhibit a number of advantages, including simple synthesis, wide variety, simple modulation of pore size, large specific surface area, excellent chemical stability, and easy modification. In recent years, numerous scientists have investigated MOFs related materials involved with organic arsenic. These studies can be divided into three categories: detection of organic arsenic by MOFs, adsorption to remove organic arsenic by MOFs, and catalytic removal of organic arsenic by MOFs. Here, we conduct a critical analysis of current research findings and knowledge pertaining to the structural characteristics, application methods, removal properties, interaction mechanisms, and spectral analysis of MOFs. We summarized the application of MOFs in organic arsenic detection, adsorption, and catalytic degradation. Other arsenic removal technologies and conventional substances are also being investigated. This review will provide relevant scientific researchers with references.


Reply: "Letter to the editor Re: Diaz M., et al. Nutrients 2018, 10, 1481".
The objective of this letter of reply is to provide answers to the doubts and critical issues that Martín Martinez and López Liñan [...].


The conformation of cross-linked actin.S-1 in the presence and absence of ATP.
Electron microscopy studies have shown that the structure of the complex of myosin subfragment 1 (S-1) cross-linked to actin with 1-ethyl-3-[3-(dimethyl-amino) propyl] carbodiimide is very different in the presence and absence of ATP (Craig, R., Greene, L. E., and Eisenberg, E. (1985) Proc. Natl. Acad. Sci. U. S. A. 82, 3247-3251). More recent studies have found that the structure of the cross-linked complex between S-1 modified extensively with N-ethylmaleimide (NEM.S-1) and actin resembles that of the rigor complex both in the presence and absence of ATP, whereas the structure of the cross-linked complex between S-1 modified with N',N'-p-phenylenedimaleimide (pPDM.S-1) and actin resembles that of the cross-linked actin.S-1 complex in the presence of ATP. In the present study, we have obtained biochemical evidence supporting these results. The conformation of the different cross-linked actin.S-1 complexes was determined by studying their effect on the troponin-tropomyosin-actin complex (regulated actin). The basis of this probe for conformation is that S-1.ATP, which is in the weak-binding conformation, interacts very differently with regulated actin than S-1 or S-1.ADP, which are in the strong-binding conformation. We find that both in the presence and absence of ATP, cross-linked NEM.S-1 appears to be in the strong-binding conformation, whereas cross-linked pPDM.S-1 appears to be shifted toward the weak-binding conformation. In contrast, cross-linked unmodified S-1 appears to be in the strong-binding conformation in the presence of ADP and the weak-binding conformation in the presence of ATP. Therefore, in agreement with electron microscopy studies, the cross-linked actin.S-1 complex appears to be able to alternate between the weak-binding and strong-binding conformation during the cross-bridge cycle.


Product of the cellular oncogene, c-fos, observed in mouse and human tissues using an antibody to a synthetic peptide.
The transforming gene of the osteosarcoma-producing FBJ murine sarcoma virus, v-fos, is homologous to a normal cellular gene, c-fos, in vertebrate species. Transcripts from the c-fos proto-oncogene accumulate to very high levels in late gestational mouse and human extra-embryonic tissues. We now report that these RNA transcripts are translated in these tissues. Rabbits were immunized with a synthetic peptide whose sequence is common to both c-fos and v-fos. After affinity purification on an immunosorbent containing the fos peptide (a nonapeptide), the antibody reacted with a component(s) in nuclei in sections of human and murine tissues and immunoprecipitated the v-fos gene product (p55) and a cellular protein of 39 kd (p39, complexed with fos) from lysates of metabolically-labelled virally transformed cells. Crude extracts of normal tissues contained major anti-fos-reactive proteins in the range of 55-60 kd as shown by protein blot analysis. Indirect immunofluorescence and immunoperoxidase staining showed that in addition to strong immunoreactive component(s) in the nuclei of extra-embryonic tissues of human and mouse, weaker reactions are detectable in all normal fetal and adult tissues tested. This demonstrates that fos-reactive protein is expressed in a wide variety of cells and tissues.


Response scale effects and integration processes in the averaging of motor movements.
In two experiments Ss averaged pairs of fixed, linear motor movements. A production method of motor averaging was used in Exp. 1, and category scaling procedures were used in Exp. 2. Stimulus interaction effects were found in Exp. 1 but not in Exp. 2. Prior exposure to long or to short movements produced slight contrast effects in each experiment, but the nature of these effects differed between experiments. Results were interpreted within the context of averaging models of information integration theory. The implications of these models for theories of motor retention were discussed.


Hemodynamic evaluation by endoscopic ultrasonography of esophageal varices resistant to injection sclerotherapy.
Some esophageal variceal cases are resistant to endoscopic injection sclerotherapy (EIS) using 5% ethanolamine oleate (5% EO). We evaluated the hemodynamics of esophageal varices that were resistant to EIS using 5% EO. Selected for this study were 290 consecutive patients who underwent hemodynamic evaluation using endoscopic color Doppler ultrasonography (ECDUS) and an ultrasonic microprobe (UMP) before EIS. EIS was performed using 5% EO with iopamidol (5% EOI) under fluoroscopy. We retrospectively evaluated the hemodynamic differences between patients resistant to and not resistant to EIS using 5% EOI. Nine patients were resistant to EIS using 5% EOI (group A). Various parameters were compared between the 281 patients who had been given EIS using 5% EOI for esophageal varices (group B) and the 9 patients in group A. The mean number of EIS treatments until shrinkage of esophageal varices was achieved in group A (6.8 ± 3.4) was significantly greater than that in group B (4.4 ± 2.1) (P < 0.01). The mean amount of 5% EOI used in group A (31.1 ± 17.4 ml) was significantly larger than that used in group B (14.9 ± 8.8 ml) (P < 0.001). The mean frequency shift of esophageal varices in group A (452.9 ± 106.6 Hz) was significantly higher than that in group B (313.0 ± 103.2 Hz) (P < 0.001) as determined by ECDUS. The mean diameter of esophageal varices as found by UMP was 8.0 ± 3.5 mm in group A and 4.6 ± 2.4 mm in group B, with the difference being significant (P < 0.01). Perforating veins inflowing from extramural to intramural regions were recognized in 8 (88.9%) of the 9 patients in group A and in 67 (24.1 %) of the 281 patients in group B. The mean diameter of the perforating vein was 3.8 ± 1.9 mm in group A and 2.1 ± 0.5 mm in group B, as shown by UMP, a difference that was statistically significant (P < 0.01). Hemodynamic evaluation revealed that the esophageal varices were of a higher grade in group A than in group B. By using ECDUS and UMP, this study shed light on the hemodynamics of esophageal variceal cases resistant to EIS using 5% EOI.


Differences in cytochalasin D-induced surface alterations between chronic lymphocytic leukaemic and normal lymphocytes.
Cytochalasin D (CD) causes an unusual surface alteration in normal lymphocytes consisting of the formation of focal irregular club-shaped cell processes. Lymphocytes from chronic lymphocytic leukaemic cases did not show this change on exposure to CD. There was either no surface change or, in some cases, clear double-membrane lined vesicles were formed and appeared to be discharged from the cell. This difference in response may be related to the changes in cell membranes known to occur in malignant transformation.


Light- and electron microscopic investigations of pulmonary tissue after high-frequency positive-pressure ventilation (HFPPV).
Narcotisized dogs were artificially ventilated for periods of two to five hours with HFPPV. During this time the blood gases, pH and bases were controlled. In spite of sufficient oxygenation and CO2-elimination, a metabolic acidosis developed which could not be fully compensated by the addition of buffer solutions. In light and electron microscopy these lungs did not differ significantly from control lungs. Haemorrhages or atelectases were never observed. Type I cells as well as Type II cells in the alveoli are unchanged, i.e. the Type II cells were not depleted but contained numerous typical lamellar bodies with a diameter of 0.4-1 mu. The blood gas barrier was not widened and was ca. 3000 A wide. The alveolar surface was coated by an often fragmented electron-dense film (monolayer of the surfactant).


Genomic structure and chromosomal location of the mouse pre-T-cell receptor alpha gene.
The mouse pre-T-cell receptor alpha (pT alpha) chain is a 33,000 M(r) glycoprotein expressed on the surface of immature thymocytes as a disulfide-linked heterodimer with the T-cell receptor beta (TCR beta) chain, and in association with proteins of the CD3 complex. The cDNA for pT alpha, isolated previously, encodes a type I transmembrane protein that is a member of the immunoglobulin (Ig) superfamily. Here we report the complete nucleotide sequence, the exon/intron structure, and the chromosomal location of the pTa gene. The gene spans about 8.4 kilobases (kb) and consists of four exons. Exon 1 encodes the 5' untranslated region, the leader peptide, and the first three amino acids of the mature protein. This exon is followed by a relatively long intron of 4.9 kb that contains many short interspersed repeats (SINEs) of the B1 and B2 family. The second exon encodes the extracellular Ig-like domain and exon 3 with just 45 base pairs the connecting peptide (CP), including the cysteine required for heterodimer formation. A similar exon/intron structure encoding corresponding parts of the mature polypeptide is found both in the Tcra and Tcrd constant region genes. The last exon encodes the transmembrane portion, the cytoplasmic tail, and about 540 nucleotides of 3' untranslated sequence, including a B2 repetitive element. In situ hybridization maps the pTa gene to the D/E1 region of mouse chromosome 17.


Immunological sexing of mammalian semen: current status and future options.
Although numerous schemes have been devised and promoted for the putative separation of X- and Y-chromosome-bearing spermatozoa, none has gained widespread acceptance. We have reexamined the possibility that Y-chromosome-bearing spermatozoa express a male-specific antigen on their plasma membrane, but that the same antigen is absent from X-chromosome-bearing spermatozoa. Immunohistochemical studies with high titer serologically detectable, male-specific antiserum revealed that serologically detectable, male-specific antigen is present on approximately 50% of spermatozoa and is located on both the postacrosomal region of the head and the midpiece of the flagellum. Similar results have also been described by other investigators working with bull spermatozoa. These data renew the question of whether the expression of serologically detectable, male-specific antigen on spermatozoa is a result of Y-chromosome-specific haploid gene expression. If so, then serologically detectable, male-specific antigen could possibly be targeted as a means of immunologically separating X- and Y-chromosome-bearing spermatozoa.


Assessment of mucosal underdosing in larynx irradiation.
Mucosal underdosing as a result of electron disequilibrium at the air cavity may affect local recurrence rates for T1 and T2 larynx cancers. Secondary build-up properties of high-energy beams have been demonstrated in a slab phantom. It was the aim of this investigation to determine whether significant surface underdosing exists for the mucosa under clinical conditions. Measurements were made using a thermoluminescent dosimetry (TLD) extrapolation technique in an anatomic larynx phantom. The larynx phantom was constructed using tissue and cartilage equivalent material, based on patient cross-sectional anatomy. Three different thicknesses of LiF ribbons, 0.14, 0.39, and 0.89 mm, were placed reproducibly at 12 different positions at the anterior, posterior, and lateral walls on the endolarynx surface. Measured doses were plotted and an extrapolation was made back to the mucosal depth to obtain the dose received at each of the positions. Results were obtained for two different field configurations, opposed laterals and oblique fields, for 6-MV X rays and opposed lateral fields from a telecesium unit. In addition, the larynx surface doses of field sizes from 4 x 6 cm2 to 7 x 6 cm2 were investigated. Surface underdosing was observed owing to the secondary build-up and build-down effect of the air cavity, and the dose measured for the three extrapolation TLDs at any position varied by up to 18%. An average variation of 6% was observed. The surface underdosing was most apparent for the 6-MV opposed lateral beam technique, where mucosa doses down to 76% of the prescribed dose were observed. Mucosal underdosing at the measurement positions was less marked with oblique techniques, telecesium treatment, and increasing field size. Because of underdosing, some surface positions receive < 80% of the prescribed dose. This may contribute to the potential for higher recurrence rates observed with high-energy photons.


Endothelial dysfunction markers and immune response indices in cosmonauts' blood after long-duration space flights.
Space flight factors are known to cause a malfunction in the human immune system and lead to damage to blood vessels. The hemostatic function of endothelium during space missions and its interaction with human immunity has not been determined so far. In this work, we investigated the markers of endothelial activation and damage (plasma concentrations of soluble thrombomodulin fraction (sTM), von Willebrand factor (vWF), highly sensitive C-reactive protein (hs-CRP)), as well as the level of D-dimer and compared them to the immunological parameters characterizing the state of human humoral and cellular immunity. The immune status of long-duration ISS crewmembers was assessed by whole-blood testing, and comprehensive postflight immune assessment included the analysis of leukocyte distribution. Flow cytometry was applied to determine the absolute counts and the percentage of lymphocyte subsets: B cells (CD19+), T cells (CD3+, CD3+CD4+, CD3+CD8+), NK cells (CD3-CD16+CD56+, CD11b+CD56+), and activated subsets (CD3+CD25+ and CD3+HLA-DR+). The in vitro basal cytokine production was investigated in whole blood cell culture. The cytokines IFN-gamma, IL-1-beta, IL-4, IL-6, IL-10, IL-18, and TNF-alpha were measured in plasma and the 24-h supernatants by a sensitive enzyme-linked immunosorbent assay. A significant increase in the plasma levels of vWF and hs-CRP and a decrease in the concentration of sTM after spaceflights were detected. Divergent changes in the parameters characterizing the state of the immune system were observed. We propose that the changes revealed may lead to an increase in the procoagulant activity of blood plasma, suppression of protein C activation and thrombin inhibition, as well as to an increase in the adhesive-aggregate potential of platelets, especially in case of changes in the rheological characteristics of blood flow during re-adaptation to ground conditions. We also speculate that the immune system might play an important role in vessel damage during long-duration missions.


Craniofacial resections for tumors involving the base of the skull.
Over a 10 year period, 42 patients with tumors involving the base of the skull underwent operation at our institution. Twenty-six patients had tumors involving the anterior fossa and cribriform plate, 3 patients had tumors involving the anterior fossa and orbit, 3 patients had invasion of the middle fossa, 5 patients had invasion of the temporal bone, and 5 patients had invasion of the clivus. A detailed analysis of the 26 patients who underwent craniofacial resection for tumors invading the anterior fossa cribriform plate region has been presented. Histologic studies revealed epithelial tumors in 18 patients, sarcoma in 6 patients, melanoma in one patient, and ossifying fibroma in one patient. The median survival in this group of patients was 60 months. Survival was influenced by histologic diagnosis. Malignant tumors involving the base of the skull can be successfully resected using a craniofacial approach with minimum morbidity and acceptable operative mortality as demonstrated in this experience. Unfavorable prognostic factors included massive intracranial extension, high grade tumor, and previous treatment failure.


Evaluation of the effect of outer primer structure, and inner primer linker sequences, in the performance of Loop-mediated isothermal amplification.
Loop-mediated isothermal amplification, or LAMP, is nowadays the most popular isothermal nucleic acid amplification technique. This technique implements a minimum of four primers, named outer (F3/B3) and inner primers (FIP/BIP). The inner primers hybridize in two distinct regions, and some studies have reported that the usage of a linker, typically composed of four thymines, in the middle of these primers can improve assay performance. In addition to this, dual-priming oligonucleotides, DPO, have been reported to provide highly specific reducing non-specific amplifications. Considering the large number of primers implemented in LAMP assays, in the current study the suitability of DPO primers replacing regular outer primers; and their combination with different linker sequences in the inner primers were explored. The results demonstrated that replacing standard F3/B3 by DPO primers does not significantly affect that overall performance of the assay, and provides additional stability to temperature changes. This observations were consistent regardless the type of linker implemented in the inner primers, out of which in the current study a linker composed of thymines significantly outperformed the other options tested, most likely due to a combination of sequence and physical structure.


Trends in cataract surgery training curricula.
To evaluate trends in cataract surgery training curricula and factors affecting timing of resident participation as a primary surgeon. Pennsylvania State College of Medicine, Hershey, Pennsylvania, USA. Cross-sectional study of anonymous survey results. A description of the study and link to an online survey was e-mailed to program directors of each ophthalmology residency training program accredited by the Accreditation Council for Graduate Medical Education (ACGME). Fifty-one (44%) of the 116 program directors completed the survey. First-year, second-year, and third-year residents performed a mean of 2, 25, and 155 phacoemulsification surgeries, respectively, as a primary surgeon. Only 1 program (2%) required residents to perform extracapsular cataract extraction (ECCE) before performing phacoemulsification. Clear corneal phacoemulsification was the first technique taught to trainees at 91% of programs. More than two thirds (71%) of program directors indicated that their program had a cataract surgery training curriculum designed to transition residents gradually to the operating room. These curricula included structured wet laboratory (92%) and lecture (89%) components. Inadequate resident knowledge and surgical skill base (57%) and anticipation of increased surgical complication risk (37%) were the most commonly reported factors impeding earlier exposure to phacoemulsification in residency. Results show that residents today begin surgical training with phacoemulsification rather than ECCE, perform a higher number of phacoemulsification surgeries than is required by the ACGME, and begin performing phacoemulsification as early as their first or second year of residency. Despite these evolutions, 29% of respondent ACGME-accredited ophthalmology residency programs reported not having a formal cataract surgery training curriculum.


A paternally imprinted QTL for mature body mass on mouse chromosome 8.
Body mass (BM) is a classic polygenic trait that has been extensively investigated to determine the underlying genetic architecture. Many previous studies looking at the genetic basis of variation in BM in murine animal models by quantitative trait loci (QTL) mapping have used crosses between two inbred lines. As a consequence it has not been possible to explore imprinting effects which have been shown to play an important role in the genetic basis of early growth with persistent effects throughout the growth curve. Here we use partially inbred mouse lines to identify QTL for mature BM by applying both Mendelian and Imprinting models. The analysis of an F2 population (n approximately 500) identified a number of QTL at 14, 16, and 18 weeks explaining in total 31.5%, 34.4%, and 30.5% of total phenotypic variation, respectively. On Chromosome 8 a QTL of large effect (14% of the total phenotypic variance at 14 weeks) was found to be explained by paternal imprinting. Although Chromosome 8 has not been previously associated with imprinting effects, features of candidate genes within the QTL confidence interval (CpG islands and direct clustered repeats) support the hypothesis that Insulin receptor substrate 2 may be associated with imprinting, but as yet is unidentified as being so.


CLN2/TPP1 deficiency: the novel mutation IVS7-10A>G causes intron retention and is associated with a mild disease phenotype.
The classical form of late infantile neuronal ceroid lipofuscinosis (LINCL) is a childhood hereditary neurodegenerative disease usually fatal in the first decade of life. The underlying gene, CLN2, encodes the lysosomal soluble enzyme tripeptidyl-peptidase 1 (TPP1). In a Portuguese patient with juvenile form of the disease, the histochemical study revealed the presence of curvilinear inclusions typical of LINCL. In vitro TPP1 activity was deficient in patient's cells. CLN2 gene analysis revealed the transition IVS7-10A>G (g.4196A>G) in both alleles. In silico analysis suggested that A-to-G change in the A-rich region of intron 7 could cause aberrant splicing of exon 8 by creating a novel acceptor splice site. However, because the wild-type acceptor of intron 7 is weak and it was not apparently affected, the severity of this mutation could not be established through sequencing data of gDNA. Normal level of spliced CLN2/mRNA was observed in patient's fibroblasts. In the cDNA, the 9-nt retention of intronic sequence (c.886_887ins9) was observed. The mutation is predicted to result in a protein with three extra amino acids between proline 295 and glycine 296. In patient's fibroblasts the level of mutant CLN2p was reduced to about 60% but the migration pattern was similar to the wild-type protein, suggesting that it was correctly targeted to the lysosomes. Taken together, these findings suggest that the first "ag" is selected for splicing and the mutant protein must retain some residual catalytic activity, thus explaining the late onset and the delayed progression of the disease.


LhGST is an anthocyanin-related glutathione S-transferase gene in Asiatic hybrid lilies (Lilium spp.).
LhGST, an anthocyanin-related GST gene, was identified from Asiatic hybrid lilies. Expression and functional analyses demonstrated that LhGST might be involved in anthocyanin sequestration in lily tepals. Anthocyanins are responsible for the pink, red and purple pigmentation of flowers in Asiatic hybrid lilies, synthesized at the cytoplasmic surface of the endoplasmic reticulum (ER) and then transported to the vacuole. To date, the mechanism involved in the intracellular transport of anthocyanins in lilies has not been well elucidated. Here, full-length glutathione S-transferase gene (LhGST) was identified from lilies. Expression analysis revealed that LhGST was positively correlated with anthocyanin accumulation. Phylogenetic tree analysis showed that LhGST clustered with other anthocyanin-related GSTs in the same phi clade. Moreover, functional complementation of an Arabidopsis tt19 mutant demonstrated that LhGST might be involved in anthocyanin accumulation in lily tepals. Additionally, according to phenotype analysis, LhGST was found to be correlated with the transport of anthocyanin in lilies by virus-induced gene silencing (VIGS) assay. In addition, cis-element analysis of the LhGST promoter showed the presence of ABA-, auxin-, MeJA-, gibberellin-, light-, and stress-responsive elements and an MYB recognition site (MRS, CCGTTG). Yeast one-hybrid and dual-luciferase report assays revealed that the promoter of LhGST was activated by LhMYB12-lat, which is a key R2R3-MYB transcription factor that regulates anthocyanin biosynthesis in lilies. In conclusion, our results revealed that LhGST plays a key role in anthocyanin transport and accumulation in the tepals of lilies.


Conscious experiences and high-density EEG patterns predicting subjective sleep depth.
What accounts for feeling deeply asleep? Standard sleep recordings only incompletely reflect subjective aspects of sleep and some individuals with so-called sleep misperception frequently feel awake although sleep recordings indicate clear-cut sleep. To identify the determinants of sleep perception, we performed 787 awakenings in 20 good sleepers and 10 individuals with sleep misperception and interviewed them about their subjective sleep depth while they underwent high-density EEG sleep recordings. Surprisingly, in good sleepers, sleep was subjectively lightest in the first 2 h of non-rapid eye movement (NREM) sleep, generally considered the deepest sleep, and deepest in rapid eye movement (REM) sleep. Compared to good sleepers, sleep misperceptors felt more frequently awake during sleep and reported lighter REM sleep. At the EEG level, spatially widespread high-frequency power was inversely related to subjective sleep depth in NREM sleep in both groups and in REM sleep in misperceptors. Subjective sleep depth positively correlated with dream-like qualities of reports of mental activity. These findings challenge the widely held notion that slow wave sleep best accounts for feeling deeply asleep. Instead, they indicate that subjective sleep depth is inversely related to a neurophysiological process that predominates in early NREM sleep, becomes quiescent in REM sleep, and is reflected in high-frequency EEG activity. In sleep misperceptors, this process is more frequently active, more spatially widespread, and abnormally persists into REM sleep. These findings help identify the neuromodulatory systems involved in subjective sleep depth and are relevant for studies aiming to improve subjective sleep quality.


Factors influencing the assay of Inoue-Melnick virus.
We describe factors influencing the assay of Inoue-Melnick virus (IMV) and practical comments for its assay. MRC-5 cells at a population doubling level (PDL) of more than 32 were more sensitive than younger cells for demonstrating cytopathic effect (CPE) of the virus. The PDL of MRC-5 cells also influenced the virus yield, particularly when the amount of viral inoculum was small. Cells at younger PDL produced less virus. The appearance of CPE and the virus yield were inhibited by increasing the concentration of NaHCO3 to 0.14% in the maintenance medium. The release of the virus into the medium was also inhibited as NaHCO3 was increased.


CDKN2A deletions in acute lymphoblastic leukemia of adolescents and young adults: an array CGH study.
Deletion in chromosome 9p involving the CDKN2A locus (9p21.3) is known in many malignancies. To detect this deletion in adolescent ALL patients we used oligo array CGH and studied 54 patients aged 10-25 years. Deletion rate was 25/54 (46%), of these 19/25 (76%) were homozygous. Small deletions (<200 kb) were found in 8/25 (32%) and the smallest deletion was <30 kb. The only gene affected in all deletions was CDKN2A. We were unable to demonstrate prognostic value of the deletion, however patients with deletion belonged more often (P=0.06) to unfavorable biological category. Our results indicate that CDKN2A deletions <200 kb may not be detected by conventional methods.


Donor Heart Treatment With COMP-Ang1 Limits Ischemia-Reperfusion Injury and Rejection of Cardiac Allografts.
The major cause of death during the first year after heart transplantation is primary graft dysfunction due to preservation and ischemia-reperfusion injury (IRI). Angiopoietin-1 is a Tie2 receptor-binding paracrine growth factor with anti-inflammatory properties and indispensable roles in vascular development and stability. We used a stable variant of angiopoietin-1 (COMP-Ang1) to test whether ex vivo intracoronary treatment with a single dose of COMP-Ang1 in donor Dark Agouti rat heart subjected to 4-h cold ischemia would prevent microvascular dysfunction and inflammatory responses in the fully allogeneic recipient Wistar Furth rat. COMP-Ang1 reduced endothelial cell-cell junction disruption of the donor heart in transmission electron microscopy during 4-h cold ischemia, improved myocardial reflow, and reduced microvascular leakage and cardiomyocyte injury of transplanted allografts during IRI. Concurrently, the treatment reduced expression of danger signals, dendritic cell maturation markers, endothelial cell adhesion molecule VCAM-1 and RhoA/Rho-associated protein kinase activation and the influx of macrophages and neutrophils. Furthermore, COMP-Ang1 treatment provided sustained anti-inflammatory effects during acute rejection and prevented the development of cardiac fibrosis and allograft vasculopathy. These results suggest donor heart treatment with COMP-Ang1 having important clinical implications in the prevention of primary and subsequent long-term injury and dysfunction in cardiac allografts.


Human VDAC pseudogenes: an emerging role for VDAC1P8 pseudogene in acute myeloid leukemia.
Voltage-dependent anion selective channels (VDACs) are the most abundant mitochondrial outer membrane proteins, encoded in mammals by three genes, VDAC1, 2 and 3, mostly ubiquitously expressed. As 'mitochondrial gatekeepers', VDACs control organelle and cell metabolism and are involved in many diseases. Despite the presence of numerous VDAC pseudogenes in the human genome, their significance and possible role in VDAC protein expression has not yet been considered. We investigated the relevance of processed pseudogenes of human VDAC genes, both in physiological and in pathological contexts. Using high-throughput tools and querying many genomic and transcriptomic databases, we show that some VDAC pseudogenes are transcribed in specific tissues and pathological contexts. The obtained experimental data confirm an association of the VDAC1P8 pseudogene with acute myeloid leukemia (AML). Our in-silico comparative analysis between the VDAC1 gene and its VDAC1P8 pseudogene, together with experimental data produced in AML cellular models, indicate a specific over-expression of the VDAC1P8 pseudogene in AML, correlated with a downregulation of the parental VDAC1 gene.


Circulating S-Glutathionylated cMyBP-C as a Biomarker for Cardiac Diastolic Dysfunction.
Background cMyBP-C (Cardiac myosin binding protein-C) regulates cardiac contraction and relaxation. Previously, we demonstrated that elevated myocardial S-glutathionylation of cMyBP-C correlates with diastolic dysfunction (DD) in animal models. In this study, we tested whether circulating S-glutathionylated cMyBP-C would be a biomarker for DD. Methods and Results Humans, African Green monkeys, and mice had DD determined by echocardiography. Blood samples were acquired and analyzed for S-glutathionylated cMyBP-C by immunoprecipitation. Circulating S-glutathionylated cMyBP-C in human participants with DD (n=24) was elevated (1.46±0.13-fold, P=0.014) when compared with the non-DD controls (n=13). Similarly, circulating S-glutathionylated cMyBP-C was upregulated by 2.13±0.47-fold (P=0.047) in DD monkeys (n=6), and by 1.49 (1.22-2.06)-fold (P=0.031) in DD mice (n=5) compared with the respective non-DD controls. Circulating S-glutathionylated cMyBP-C was positively correlated with DD in humans. Conclusions Circulating S-glutathionylated cMyBP-C was elevated in humans, monkeys, and mice with DD. S-glutathionylated cMyBP-C may represent a novel biomarker for the presence of DD.


Aging effects on visual evoked potentials (VEPs) for motion direction discrimination.
Age-related declines in motion perception have been well documented. We investigated the impact of age on electrophysiological correlates of motion perception, namely the P1 and N2 components of motion onset visual evoked potentials (MO-VEPs). Additionally, we used a model of response times based on the diffusion model to pinpoint the cognitive processes affected by aging. Twelve healthy adults (age <55 years) and 19 elderly (age >55 years) performed a motion direction discrimination task during EEG recording. Behaviorally, younger and older participants had similar, high accuracy rates--98% correct, but older adults exhibited 85 ms longer response times. Fitting behavioral results with a diffusion model revealed differences between young adults and elderly in non-decision time, which we argue reflects an early perceptual stage. Electrophysiologically, aging effects were present at MO-VEPs P1 and N2 components at the posterior sites. For the P1 component, older as compared to younger adults showed greater topographical voltage distribution. For the N2 component of elderly as compared to young adults we found delayed onsets and diminished amplitudes. We did not find any significant correlations between behavioral and MO-VEP measures. However, regression analysis showed that N2 amplitude and latency were significant age predictors. Overall, our results indicate that in motion perception, age-related changes occur in early stages of visual processing, most likely in striate and extrastriate visual cortices.


Back muscle activity and sagittal spinal alignment during quadruped upper and lower extremity lift in young men with low back pain history.
Quadruped upper and lower extremity lift (QULEL) is performed for selective training of the lumbar multifidus muscle in patients with low back pain (LBP) or individuals with LBP history (LBPH). However, the activities of the back muscles and sagittal spinal alignment during QULEL are not clarified in individuals with LBPH. This study aimed to analyze the activities of the back muscles and sagittal spinal alignment during QULEL in young male with LBPH. The study comprised 9 asymptomatic young men and 8 young men with LBPH. The activities of the lumbar multifidus, latissimus dorsi and thoracic erector spinae, and lumbar erector spinae muscles were measured using surface electromyography. The flexion angles of the upper and lower thoracic spine, and extension angle of the lumbar spine were measured using a 6-DF electromagnetic motion tracking system. The association with LBPH was investigated using multiple logistic regression analysis with a forward selection method, with the activities of the back muscles, sagittal spinal alignment, age, body height, and body weight as independent variables. Multiple logistic regression analysis (p = 0.0002) showed that the activity of the latissimus dorsi and thoracic erector spinae muscles in the side on which the lower extremity was lifted and body height were significant and independent determinants of LBPH, but other factors were not. The results of this study suggest that the activity of the latissimus dorsi and thoracic erector spinae muscles increases while there are no decrease in activity of the lumbar multifidus muscle and excessive extension of the lumbar spine during QULEL in young men with LBPH.


The prevalence and incidence of coronary heart disease is significantly increased in periodontitis: a meta-analysis.
Previous studies have shown conflicting results as to whether periodontitis (PD) is associated with increased risk of coronary heart disease (CHD). The aim of the current study was to evaluate whether such an association exists. A systematic review of the literature revealed 5 prospective cohort studies (follow-up >6 years), 5 case-control studies, and 5 cross-sectional studies that were eligible for meta-analysis. Individual studies were adjusted for confounding factors such as age, sex, diabetes mellitus, and smoking. The 3 study categories were analyzed separately. Heterogeneity of the studies was assessed by Cochran Q test. The studies were homogeneous; therefore, the Mantel-Haenszel fixed-effect model was used to compute common relative risk and odds ratio (OR). Meta-analysis of the 5 prospective cohort studies (86092 patients) indicated that individuals with PD had a 1.14 times higher risk of developing CHD than the controls (relative risk 1.14, 95% CI 1.074-1.213, P < .001). The case-control studies (1423 patients) showed an even greater risk of developing CHD (OR 2.22, 95% CI 1.59-3.117, P < .001). The prevalence of CHD in the cross-sectional studies (17724 patients) was significantly greater among individuals with PD than in those without PD (OR 1.59, 95% CI 1.329-1.907, P < .001). When the relationship between number of teeth and incidence of CHD was analyzed, cohort studies showed 1.24 times increased risk (95% CI 1.14-1.36, P < .0001) of development of CHD in patients with <10 teeth. This meta-analysis indicates that both the prevalence and incidence of CHD are significantly increased in PD. Therefore, PD may be a risk factor for CHD. Prospective studies are required to prove this assumption and evaluate risk reduction with the treatment of PD.


Conization, frozen section examination, and planned hysterectomy in the treatment of high-grade cervical intraepithelial neoplasia.
We tested the role of frozen section examination of the cone specimen in the evaluation of the resection margin status and to rule out invasion in patients with high-grade cervical intraepithelial neoplasia. Twenty-five patients with cervical intraepithelial neoplasia underwent conization followed by frozen section examination and planned hysterectomy. The results of the definitive paraffin exam were compared with frozen section examination. In the evaluation of the margins by frozen section examination, 16 patients (64%) had positive cone margins and 9 (36%) had negative margins. The definitive paraffin examination of margin status was concordant in all the cases. Intraoperative diagnosis of invasion was made in 5 cases, and 1 of these was microinvasive. Among the remaining 20 cases, we detected 2 additional microinvasive carcinomas after paraffin study, so the diagnosis of the frozen section examination was concordant with the paraffin sections in 23/25 cases (92%). Two cases of microinvasive carcinoma were diagnosed as cervical intraepithelial neoplasia by frozen section examination and had less than 2 mm stromal invasion. In high-grade cervical intraepithelial neoplasia, frozen section examination can provide immediate and precise evaluation of the cone margin status in high-grade cervical intraepithelial neoplasia. It can identify frank invasion and permit adequate treatment in a one-stage procedure. In early microinvasive disease, frozen section examination fails to detect the area of invasion but reliably detects clear resection margins.


The differences between fungal α-glucan synthase determining pullulan synthesis and that controlling cell wall α-1,3 glucan synthesis.
The fungal α-glucan synthases (Agss) are multi-domain proteins catalyzing biosynthesis of cell wall α-1,3-glucan which determines cell wall integrity or fungal pathogenicity and pullulan which is a maltotriosyl polymer made of α-1,4 and α-1,6 bound glucose units. The Agss family can be divided into 11 groups, some of which lost the original functions due to accumulation of harmful mutations or gene loss. Schizosaccharomyces pombe kept five kinds of Agss in the genome while Aspergillus spp. and Penicillium spp. lost one or two or three kinds of Agss. All the human, animal and plant pathogens kept only one single kind of Ags or only one active Ags for synthesis of cell wall α-1,3-glucan, a virulence factor. While the genus Aureobasidium spp. contained three kinds of Agss, of which only some of the Ags2 was involved in pullulan biosynthesis. Although many Agss contained Big_5 domain, only the Big_5 domain with conserved amino acids LQS from some strains of A. melanogenum could catalyze pullulan biosynthesis. This whole amino acid sequence and phylogenetic differences may cause non-α-1,3-glucan synthesizing activity of some fungal Agss.


Hydroxyfasudil ameliorates bladder dysfunction in male spontaneously hypertensive rats.
To investigate the effect of hydroxyfasudil, a nonselective Rho-kinase inhibitor, on hypertension-related bladder dysfunction in spontaneously hypertensive rats (SHRs), as there is an increasing evidence that the Rho-associated protein kinase (ROCK) system plays an important role in bladder contraction. Twelve-week-old male SHRs were treated with hydroxyfasudil (1 mg/kg i.p.) once a day for 6 weeks. Wistar rats and SHRs without treatment with hydroxyfasudil were used as controls. Six weeks after the hydroxyfasudil treatment, voiding functions were evaluated by metabolic cages and cystometric studies under urethane anesthesia (1.0 g/kg i.p.). Bladder blood flow (BBF) was estimated by the hydrogen clearance method. The bladder tissue levels of nerve growth factor (NGF) and ROCK activity were evaluated by the ELISA method. The SHR showed significant increases in micturition frequency and decreases in single-voided volume in metabolic cages, and significantly increases in micturition frequency and intercontractile interval in cystometric studies. Furthermore, the SHR showed significant increases in the BBF and bladder NGF concentration compared with the Wistar rats. These alterations in the SHR were significantly ameliorated after treatment with hydroxyfasudil, with small changes in blood pressure. However, the maximum detrusor pressures during voiding and the ROCK activities in the experimental bladders were similar in all rats examined. Our data indicate that this dose of hydroxyfasudil was effective on the BBF, whereas it had no significant effect on micturition pressure. These findings suggest that hydroxyfasudil could ameliorate hypertension-related bladder dysfunction in the SHR via improvement of the BBF (248 words).


Characterizing T2 iso- and hypo-intense renal masses on MRI: Can templated algorithms improve accuracy?
To assess if a templated algorithm can improve the diagnostic performance of MRI for characterization of T2 isointense and hypointense renal masses. In this retrospective study, 60 renal masses with histopathologic diagnoses that were also confirmed as T2 iso- or hypointense on MRI were identified (mean ± standard deviation, range: 3.9 ± 2.5, 1.0-13.7 cm). Two semi-quantitative diagnostic algorithms were created based on MRI features of renal masses reported in the literature. Three body-MRI trained radiologists provided clinical diagnoses based on their experience and separately provided semiquantitative data for each components of the two algorithms. The algorithms were applied separately by a radiology trainee without additional interpretive input. Logistic regression was used to compare the accuracy of the three methods in distinguishing malignant versus benign lesions and in diagnosing the exact histopathology. Inter-reader agreement for each method was calculated using Fleiss' kappa statistics. The accuracy of the two algorithms and clinical experience were similar (70%, 69%, and 64%, respectively, p = 0.22-0.32), with fair to moderate inter-reader agreement (Fleiss's kappa: r = 0.375, r = 0.308, r = 0.375, respectively, all p < 0.0001). The accuracy of the two algorithms and clinical experience in diagnosing specific histopathology were also no different from each other (34%, 29%, and 32%, respectively, p = 0.49-0.74), with fair to moderate inter-reader agreement (Fleiss's kappa: r = 0.20, r = 0.28, r = 0.375, respectively, all p < 0.0001). Semi-quantitative templated algorithms based on MRI features of renal masses did not improve the ability to diagnose T2 iso- and hypointense renal masses when compared to unassisted interpretation by body MR trained subspecialists.


De novo assembly and characterization of the leaf, bud, and fruit transcriptome from the vulnerable tree Juglans mandshurica for the development of 20 new microsatellite markers using Illumina sequencing.
Manchurian walnut (Juglans mandshurica Maxim.) is a vulnerable, temperate deciduous tree valued for its wood and nut, but transcriptomic and genomic data for the species are very limited. Next generation sequencing (NGS) has made it possible to develop molecular markers for this species rapidly and efficiently. Our goal is to use transcriptome information from RNA-Seq to understand development in J. mandshurica and develop polymorphic simple sequence repeats (SSRs, microsatellites) to understand the species' population genetics. In this study, more than 47.7 million clean reads were generated using Illumina sequencing technology. De novo assembly yielded 99,869 unigenes with an average length of 747 bp. Based on sequence similarity search with known proteins, a total of 39,708 (42.32 %) genes were identified. Searching against the Kyoto Encyclopedia of Genes and Genomes Pathway database (KEGG) identified 15,903 (16.9 %) unigenes. Further, we identified and characterized 63 new transcriptome-derived microsatellite markers. By testing the markers on 4 to 14 individuals from four populations, we found that 20 were polymorphic and easily amplified. The number of alleles per locus ranged from 2 to 8. The observed and expected heterozygosity per locus ranged from 0.209 to 0.813 and 0.335 to 0.842, respectively. These twenty microsatellite markers will be useful for studies of population genetics, diversity, and genetic structure, and they will undoubtedly benefit future breeding studies of this walnut species. Moreover, the information uncovered in this research will also serve as a useful genetic resource for understanding the transcriptome and development of J. mandshurica and other Juglans species.


Adaptive biomimicry: design of neural interfaces with enhanced biointegration.
Neural interfaces (NIs) have traditionally used inorganic device constructs paired with electrical stimulation to bypass injured or diseased electroactive tissues. These bioinert devices have significant impact on the neural tissue, being synthetic and causing large volumetric changes to the biological environment. The concept of biomimicry has become popular for tissue engineering technologies, reflecting biological properties as a component of material design. Tissue engineering strategies can be harnessed in bioelectronic device design to improve biological tolerance, but the need for improved integration with the native tissue remains an unmet need. Adaptive biomimetic designs that respond to the changing neural tissue environment associated with wound healing can actively address the immune response to improve biointegration. These adaptive approaches include responsive materials paired with stem cells and bioactive molecules as integrated components of NIs. Combining adaptive biomimetics with NIs provides a new, more natural approach for communicating with the nervous system.


A scanning electron microscopic study of arterial endothelial cells using vascular casts.
Vascular casts were made of rabbit aortas by infusing Batson's No. 17 anatomical corrosion compound into the artery at physiological pressure. The arterial tissue was then digested with sodium hydroxide and the cast viewed by scanning electron microscopy (SEM). Outlines of the endothelial cells and their silver stained boundaries were clearly visible. Cell nuclei and fine surface detail were also discernible. In EDTA damaged arteries, injured endothelial cells and platelets could also be observed in the vascular casts.


Purification and properties of thiosulfate reductase from Desulfovibrio gigas.
Thiosulfate reductase of the dissimilatory sulfate-reducing bacterium Desulfovibrio gigas has been purified 415-fold and its properties investigated. The enzyme was unstable during the different steps of purification as well as during storage at - 15 degrees C. The molecular weight of thiosulfate reductase estimated from the chromatographic behaviour of the enzyme on Sephadex G-200 was close to 220000. The absorption spectrum of the purified enzyme exhibited a protein peak at 278 nm without characteristic features in the visible region. Thiosulfate reductase catalyzed the stoichiometric production of hydrogen sulfide and sulfite from thiosulfate, and exhibited tetrathionate reductase activity. It did not show sulfite reductase activity. The optimum pH of thiosulfate reduction occurred between pH 7.4 and 8.0 and its Km value for thiosulfate was calculated to be 5 - 10(-4)M. The sensitivity of thiosulfate reductase to sulfhydryl reagent and the reversal of the inhibition by cysteine indicated that one or more sulfhydryl groups were involved in the catalytic activity. The study of electron transport between hydrogenase and thiosulfate reductase showed that the most efficient coupling was obtained with a system containing cytochromes c3 (Mr = 13000) and c3 (Mr = 26000).


Effectiveness of sorting calves with high risk of developing bovine respiratory disease on the basis of serum haptoglobin concentration at the time of arrival at a feedlot.
To evaluate serum haptoglobin concentration at feedlot arrival and subsequent performance and morbidity and mortality rates of calves that developed bovine respiratory disease. 360 heifer calves and 416 steer and bull calves. Serum samples were obtained from cattle at the time of arrival to a feedlot (day -1) and analyzed for haptoglobin concentration. In experiment 1, calves were classified into groups with a low (< 1.0 μg/mL), medium (1.0 to 3.0 μg/mL), or high (> 3.0 μg/mL) serum haptoglobin concentration and allotted into pens on the basis of group. In experiment 2, calves were classified as having or not having detectable serum haptoglobin concentrations. In experiment 1, average daily gain from days 1 to 7 decreased as haptoglobin concentration increased. Dry-matter intake (DMI) from days 1 to 21 decreased with increasing haptoglobin concentration, and DMI typically decreased from days 1 to 63. Total bovine respiratory disease morbidity rate typically increased with increasing haptoglobin concentration. At harvest, no differences in carcass characteristics were observed on the basis of haptoglobin concentration. In experiment 2, cattle with measureable serum haptoglobin concentrations at arrival weighed less throughout the experiment, gained less from days 1 to 7, and had lower DMI from days 1 to 42. Overall morbidity rate was not different between groups, but cattle with detectable serum haptoglobin concentrations had higher odds of being treated 3 times. Serum haptoglobin concentration in cattle at the time of feedlot arrival was not associated with overall performance but may have limited merit for making decisions regarding targeted prophylactic treatment.


Nerve root injuries in patients with chronic low back pain.
In conclusion, the nerve roots and the DRG play an important role in the pain mechanisms of patients suffering from chronic low back pain. Signs of demyelination and increased sensitization for stimuli occurs after a direct nerve root trauma, and the plasticity for the DRG also may change the response to a given peripheral stimuli when repeated frequently over a long period of time. The regeneration mechanisms of spinal nerve roots and DRG regarding function are slow, and the final grade of recurrence depends on the degree of injury. The limited regeneration mechanisms for nerve injury and the fact that "established chronic pain centers" are hard to influence after a long pain history favor an aggressive strategy for pain management. Today, a number of treatment strategies exist for chronic low back pain patients (with or without a diagnosed nerve root injury). These strategies include physiotherapy, nonsteroid anti-inflammatory drugs (NSAIDs), steroids, analgesics of different types and administration routes, surgery, and other sorts of invasive treatments. Further knowledge about the nerve root, DRG, and the rest of the nervous system in these patients is necessary; for understanding how and when to treat patients with chronic low back pain, we need to understand more about what we are trying to treat.


Fipronil in eggs. Is LC-MS/MS the only option? A comparison study of LC-MS/MS and GC-ECD for the analysis of fipronil.
Fipronil is a broad-spectrum insecticide belonging to the phenylpyrazole chemical family, classified by the U.S. EPA as a group C (possible human) carcinogen. It is highly toxic to crustaceans, insects and zooplankton as well as to termites, rabbits and certain groups of gallinaceous birds. In European Union and Greece, fipronil is authorized only for the control of termites and cockroaches and not as a plant protection product or as a veterinary drug. The definition of residue of fipronil comprises of the sum of fipronil and its sulfone metabolite expressed as fipronil and is set at 5 μg kg-1. In this study, a sensitive and reliable modified QuEChERS method, proposed by the European Reference Laboratory for animal origin products, has been validated and applied to the residue analysis of fipronil and its metabolites (sulfone and desulfinyl) in chicken eggs and other egg products by LC-MS/MS and GC-ECD analysis. The two analytical systems performed almost equally concerning validation process and with regard to the analysis of real samples, results from both systems were in agreement: out of 11 samples analysed, 4 samples showed measurable residues of fipronil sulfone with 3 samples exceeding the MRL limit.


From microbial community structure to metabolic inference using paprica.
Microbial taxonomic marker gene studies using 16S rRNA gene amplicon sequencing provide an understanding of microbial community structure and diversity; however, it can be difficult to infer the functionality of microbes in the ecosystem from these data. Here, we show how to predict metabolism from phylogeny using the paprica pipeline. This approach allows resolution at the strain and species level for select regions on the prokaryotic phylogenetic tree and provides an estimate of gene and metabolic pathway abundance. For complete details on the use and execution of this protocol, please refer to Erazo and Bowman (2021).


Stability studies on chemoembolization mixtures. Dialysis studies of doxorubicin and lipiodol with Avitene, Gelfoam, and Angiostat.
Chemoembolization, using a combination of embolic and chemotherapeutic agents, appears to be an effective treatment for hepatocellular carcinoma. Although the postulated mechanism of effectiveness hinges on a prolonged drug delivery, increasing evidence suggests that embolization mixtures are not stable. The objective of this study was to investigate examples of these mixtures. Dialysis techniques have been used to examine the pharmacokinetic properties of chemoembolization mixtures that contain doxorubicin, Lipiodol (Guerbet Products, Montreal, Quebec), and the embolizing agents Avitene (Alcon Laboratories Inc., Fort Worth, Texas), Gelfoam (Upjohn, Kalamazoo, MI), and Angiostat (Regional Therapeutic Inc., Pacific Palisades, CA). Lipiodol, Gelfoam, and Avitene, when combined with doxorubicin, had only a small effect on the diffusion of the drug when compared with the diffusion curve of doxorubicin alone. Gelfoam or Avitene produced a thrombus-like consistency when added to a doxorubicin/Lipiodol combination, and an additional decrease in the doxorubicin appearance rate was observed. However, after 6 hours, doxorubicin levels for these mixtures reached control values observed in 3 hours. Angiostat without Lipiodol produced a profound concentration-dependent decrease in the diffusion of doxorubicin. After 9 hours, only 23% of the doxorubicin had been released. The strong complexing ability of the embolic agent Angiostat may enable it to be a carrier for doxorubicin and surpass other mixtures currently employed for transcatheter chemoembolization.


Sensorineural hearing loss in patients with inflammatory bowel disease: a subclinical extraintestinal manifestation.
Isolated case reports in which symptomatic hearing loss develops suddenly during the course of inflammatory bowel disease (IBD) have been reported, but the presence of subclinical sensorineural hearing loss (SNHL) associated with IBD has been investigated in only two preliminary studies. In order to research this further, we aimed to investigate the presence of subclinical SNHL in IBD by comparison with a control group and to examine possible relations between the bowel disease parameters and hearing loss.Otoscopy, tympanometry, and pure tone audiometry were carried out in 39 patients with IBD (21 Crohn's disease [CD], 18 ulcerative colitis [UC]) and 25 healthy age- and sex-matched controls. All patients and control subjects had normal otoscopy findings and tympanometry was unremarkable, excluding middle ear disease and conductive hearing loss. Analysis of each frequency examined showed that the average hearing thresholds were increased significantly in the study group compared to those of the control group at higher frequencies (2, 4, and 8 kHz). When these parameters were compared with the control group according to subgroups of IBD, a significant difference was determined for the UC group at frequencies of 2, 4, and 8 kHz and for the CD group only at the frequency of 4 kHz. Although there was a trend of increment in SNHL as the age of the patient and duration and extent of UC increased, no significant correlation was observed between SNHL and these parameters or sex, activity, involvement site, medication history of IBD, and coexistence of other extraintestinal manifestations. In conclusion, it was demonstrated that a subclinical SNHL may be associated with UC and somewhat with CD, affecting mainly the high frequencies. In light of this finding, it may be advisable to investigate labyrinth functions as well as other extraintestinal manifestations in patients with IBD.


Life expectancy and cardiovascular mortality in persons with schizophrenia.
To assess the impact of cardiovascular disease on the excess mortality and shortened life expectancy in schizophrenic patients. Patients with schizophrenia have two-fold to three-fold higher mortality rates compared with the general population, corresponding to a 10-25-year reduction in life expectancy. Although the mortality rate from suicide is high, natural causes of death account for a greater part of the reduction in life expectancy. The reviewed studies suggest four main reasons for the excess mortality and reduced life expectancy. First, persons with schizophrenia tend to have suboptimal lifestyles including unhealthy diets, excessive smoking and alcohol use, and lack of exercise. Second, antipsychotic drugs may have adverse effects. Third, physical illnesses in persons with schizophrenia are common, but diagnosed late and treated insufficiently. Lastly, the risk of suicide and accidents among schizophrenic patients is high. Schizophrenia is associated with a substantially higher mortality and curtailed life expectancy partly caused by modifiable risk factors.


Refsum disease. Clinical and morphological report on a case.
An atypical case of Refsum disease is reported together with the peripheral nerve morphological data. The body fluids must be assayed for phytanic acid whenever an atypical chronic peripheral neuropathy is observed.


CTLA4 protects against maladaptive cytotoxicity during the differentiation of effector and follicular CD4+ T cells.
As chronic antigenic stimulation from infection and autoimmunity is a feature of primary antibody deficiency (PAD), analysis of affected patients could yield insights into T-cell differentiation and explain how environmental exposures modify clinical phenotypes conferred by single-gene defects. CD57 marks dysfunctional T cells that have differentiated after antigenic stimulation. Indeed, while circulating CD57+ CD4+ T cells are normally rare, we found that they are increased in patients with PAD and markedly increased with CTLA4 haploinsufficiency or blockade. We performed single-cell RNA-seq analysis of matched CD57+ CD4+ T cells from blood and tonsil samples. Circulating CD57+ CD4+ T cells (CD4cyt) exhibited a cytotoxic transcriptome similar to that of CD8+ effector cells, could kill B cells, and inhibited B-cell responses. CTLA4 restrained the formation of CD4cyt. While CD57 also marked an abundant subset of follicular helper T cells, which is consistent with their antigen-driven differentiation, this subset had a pre-exhaustion transcriptomic signature marked by TCF7, TOX, and ID3 expression and constitutive expression of CTLA4 and did not become cytotoxic even after CTLA4 inhibition. Thus, CD57+ CD4+ T-cell cytotoxicity and exhaustion phenotypes are compartmentalised between blood and germinal centers. CTLA4 is a key modifier of CD4+ T-cell cytotoxicity, and the pathological CD4cyt phenotype is accentuated by infection.


Microwave-assisted continuous flow phytosynthesis of silver nanoparticle/reduced graphene oxide composites and related visible light catalytic performance.
The creation of an environmentally friendly synthesis method for silver nanomaterials (AgNPs) is an urgent concern for sustainable nanotechnology development. In the present study, a novel straightforward and green method for the preparation of silver nanoparticle/reduced graphene oxide (AgNP/rGO) composites was successfully developed through the combination of phytosynthesis, continuous flow synthesis and microwave-assistance. Oriental persimmon (Diospyros kaki Thunb.) extracts were used as both plant reducing and capping agents for fast online synthesis of AgNP/rGO composites. The experimental parameters were optimized and the morphologies of the prepared materials were investigated. The characterization results reveal that spherical AgNPs were quickly synthesized and uniformly dispersed on rGO sheets using the proposed online system. Fourier transform infrared spectroscopy analysis confirmed that phenols, flavonoids, and other substances in the plant extracts played a decisive role in the synthesis of AgNP/rGO composites. Using sodium borohydride (NaBH4) degradation of p-nitrophenol (4-NP) as a model, the catalytic activity of the prepared AgNP/rGO materials was evaluated. The complete degradation of 4-NP was achieved within 12 min through the use of AgNP/rGO materials, and the composite had a much better catalytic activity than the bare AgNPs and rGO had. Compared with the conventional chemical method, our online method is facile, fast, cost-efficient, and environmentally friendly.


SOAT1 methylation is associated with coronary heart disease.
This study was designed to investigate whether differential DNA methylationin of cholesterol absorption candidate genes can function as a biomarker for patients with coronary heart disease (CHD). DNA methylation levels of the candidate genes FLOT1, FLOT2 and SOAT1 were measured in peripheral blood leukocytes (PBLs) from 99 patients diagnosed with CHD and 89 control subjects without CHD. A total of 110 CPG sites around promoter regions of them were examined. Compared with groups without CHD, patients with CHD had lower methylation levels of SOAT1 (P<0.001). When each candidate genes were divided into different target segments, patients with CHD also had lower methylation levels of SOAT1 than patients without (P = 0.005). After adjustment of other confounders, methylation levels of SOAT1 were still associated with CHD (P = 0.001, OR = 0.290, 95% CI: 0.150-0.561). SOAT1 methylation may be associated with development of CHD. Patients with lower methylation levels in SOAT1 may have increased risks for CHD. Further studies on the specific mechanisms of this relationship are necessary.


Bilateral retinal artery and choriocapillaris occlusion following the injection of long-acting corticosteroid suspensions in combination with other drugs: I. Clinical studies.
Two well-documented cases of bilateral retinal artery and choriocapillaris occlusions with blindness following head and neck soft-tissue injection with methylprednisolone acetate in combination with lidocaine, epinephrine, or penicillin are reported. One case had only a unilateral injection. The acute observations included hazy sensorium, superior gaze palsy, pupillary abnormalities, and conjunctival hemorrhages with edema. Follow-up changes showed marked visual loss, constricted visual fields, optic nerve pallor, vascular attenuation, and chorioretinal atrophy. The literature is reviewed, and possible causes are discussed.


Intracellular lectin-binding sites in symbiont-bearing Crithidia species.
Crithidia oncopelti, C. deanei, and C. desouzai are flagellates of the Trypanosomatidae family that present bacterium-like endosymbionts in their cytoplasm. Direct and indirect lectin-gold labeling techniques were used at the electron microscopic level in Lowicryl K4M-embedded cells to demonstrate the presence of intracellular lectin-binding sites. We used the lectins Ulex europaeus I, Griffonia simplicifolia II, Ricinus communis I, Arachis hypogaea, G. simplicifolia I, Wistaria floribunda, Limulus polyphemus, and Canavalia ensiformis, which recognize alpha-L-fucose, alpha- and beta-N-acetylglucosamine, beta-galactose and beta-N-acetylgalactosamine, beta-galactose, alpha-galactose, beta-N-acetylgalactosamine, sialic acid and alpha-D-mannose, and alpha-D-glucose residues, respectively. The nucleus was the cellular structure most frequently labeled by the lectins. The Golgi complex was seldom labeled, whereas the endoplasmic reticulum and the flagellar pocket presented a large number of binding sites. Symbionts had their two unit membranes weakly labeled by the different lectins but displayed no labeling of the space between the membranes.


Comparative Efficacy of Different Drugs for the Treatment of Dilated Cardiomyopathy: A Systematic Review and Network Meta-analysis.
At present, the therapies of dilated cardiomyopathy concentrated on the symptoms of heart failure and related complications. The study is to evaluate the clinical efficacy of a combination of various conventional and adjuvant drugs in treating dilated cardiomyopathy via network meta-analysis. The study was reported according to the PRISMA 2020 statement. From inception through 27 June 2022, the PubMed, Embase, Cochrane library, and Web of Science databases were searched for randomized controlled trials on medicines for treating dilated cardiomyopathy. The quality of the included studies was evaluated according to the Cochrane risk of bias assessment. R4.1.3 and Revman5.3 software were used for analysis. There were 52 randomized controlled trials in this study, with a total of 25 medications and a sample size of 3048 cases. The network meta-analysis found that carvedilol, verapamil, and trimetazidine were the top three medicines for improving left ventricular ejection fraction (LVEF). Ivabradine, bucindolol, and verapamil were the top 3 drugs for improving left ventricular end-diastolic dimension (LVEDD). Ivabradine, L-thyroxine, and atorvastatin were the top 3 drugs for improving left ventricular end-systolic dimension (LVESD). Trimetazidine, pentoxifylline, and bucindolol were the top 3 drugs for improving the New York Heart Association classification (NYHA) cardiac function score. Ivabradine, carvedilol, and bucindolol were the top 3 drugs for reducing heart rate (HR). A combination of different medications and conventional therapy may increase the clinical effectiveness of treating dilated cardiomyopathy. Beta-blockers, especially carvedilol, can improve ventricular remodeling, cardiac function, and clinical efficacy in patients with dilated cardiomyopathy (DCM). Hence, they can be used if patients tolerate them. If LVEF and HR do not meet the standard, ivabradine can also be used in combination with other treatments. However, since the quality and number of studies in our research were limited, large sample size, multi-center, and high-quality randomized controlled trials are required to corroborate our findings.


Electrophysiologic actions and antifibrillatory efficacy of subacute left stellectomy in a conscious, post-infarction canine model of ischemic ventricular fibrillation.
The autonomic nervous system appears to modulate ventricular arrhythmias associated with acute myocardial ischemia. This study investigated the electrophysiologic effects and antifibrillatory actions of subacute left stellectomy in a conscious, post-infarction canine model of sudden cardiac death. Twenty-two dogs with a previous anterior wall myocardial infarction and inducible ventricular arrhythmias were randomized to undergo either left stellectomy (n = 12) or remain as sham-denervated controls (n = 10). Five to 7 days post left stellectomy, there were no significant changes in heart rate, electrocardiographic intervals or ventricular refractoriness compared to sham-denervated controls. Acute posterolateral ischemia was produced in left stellectomy and sham-denervated dogs by anodal current-induced thrombosis via a previously positioned electrode in the left circumflex coronary artery. Ventricular fibrillation developed within 1 hour of the onset of ischemia (early ventricular fibrillation) in 3/12 (25%) left stellectomy dogs versus 8/10 (80%) sham-denervated controls (P less than 0.05). However, 24-hour mortality rate was 5/12 (42%) after left stellectomy versus 8/10 (80%) after sham denervation (P = 0.072). Small differences in regional myocardial norepinephrine content, which is a marker for neuronal integrity, occurred in the mid-posterolateral and mid-anteroseptal regions of the left ventricle after left stellectomy. Overall norepinephrine concentration after left stellectomy was 409.70 +/- 9.90 ng/g vs 428.07 +/- 10.84 ng/g in sham controls (P = NS). In summary, subacute left stellectomy significantly reduces the incidence of ventricular fibrillation occurring within 1 hour of the onset of acute posterolateral ischemia at a distance to a previous myocardial infarction in conscious dogs, and tends to reduce the ischemic post-infarction mortality at 24 hours after the onset of ischemia. This protective effect of left stellectomy is not due to any alteration in cardiac electrophysiologic parameters measured prior to the development of acute posterolateral ischemia, nor is it related to regional denervation as determined by myocardial tissue concentration of residual norepinephrine.


Fine-tuned expression of programmed death 1 ligands in mature dendritic cells stimulated by CD40 ligand is critical for the induction of an efficient tumor specific immune response.
During maturation, murine myeloid dendritic cells (DCs) upregulated the expressions of CD11c, CD25, CD40, CD80, CD86, MHC II and programmed death 1 ligands 1 and 2 (PD-L1 and PD-L2). Differential expression patterns of PD-L1 and PD-L2 were found when DCs were triggered by CD40 ligand and TNF-alpha. PD-L1 expression was repressed and PD-L2 expression remained unchanged in mature CD40-ligated DCs, whereas TNF-alpha stimulated DCs kept high expression of PD-L1 and significantly enhanced PD-L2 expression on DCs. Proliferations of T lymphocytes stimulated by immature DCs were enhanced by blockade of the PD-1 and PD-1 ligand interaction. But inhibitive effects were found in T lymphocytes stimulated by CD40-ligated DCs. With the fine-tuned expressions of PD-L1 and PD-L2, CD40-ligated DCs could sustain a longer activation period and elicit a more efficient T lymphocyte activation.


The ability of phase angle and body composition to predict risk of death in maintenance hemodialysis patients.
The objective of this study was to investigate the ability of phase angle and body composition to identify risk factors for mortality among patients receiving maintenance hemodialysis (MHD) treatment. In this retrospective study, we examined the causes of death in 43 MHD patients who were treated at our hemodialysis center between January 2016 and December 2021 and compared the patients to 71 patients who survived during the same period. Body composition was measured using direct segmental multi-frequency bioelectrical impedance to obtain phase angle, fat-free mass (FFM), extracellular water/total body water (ECW/TBW), and waist circumference (WC). Laboratory data were also collected. Phase angle cut-off value-associated variables were identified using ROC analysis. The ability of body composition variables to identify risk factors for death in MHD patients was evaluated. We found that cardiovascular disease was the most common cause of death among MHD patients. ROC curve analysis revealed that the optimal cut-off value for phase angle as a predictor of death risk in MHD patients was 4.50°. Additionally, lower phase angle, increased age, longer dialysis vintage, lower KT/V, and hypoproteinemia were identified as significant risk factors for death in MHD patients. In conclusion, our findings suggest that cardiovascular disease is the leading cause of death among MHD patients and that lower phase angle, increased age, longer dialysis duration, and hypoproteinemia can be used to predict the risk of mortality in this patient population. The underlying mechanism by which lower phase angle can be used to predict the prognosis of MHD patients warrants further investigation.