Genome-wide DNA methylation profiling identifies epigenetic signatures of gastric cardiac intestinal metaplasia.
Measuring the DNA methylome may offer the opportunity to identify novel disease biomarkers and insights into disease mechanisms. Although aberrant DNA methylation has been investigated in many human cancers and precancerous lesions, the DNA methylation landscape of gastric cardiac intestinal metaplasia (IM) remains unknown. Therefore, we aimed to investigate the genome-wide DNA methylation landscape and to search for potential epigenetic biomarkers of gastric cardiac IM. Histopathologic profiling was performed on a total of 118 gastric cardiac biopsies from cancer-free individuals. Genome-wide DNA methylation analysis was performed on 11 gastric cardiac mucosal biopsies (IM = 7; normal = 4) using Illumina 850K microarrays. Transcriptional relevance of any candidate epigenetic biomarker was validated by qRT-PCR. The detection rate of gastric cardiac IM was 23% (27/118) in cancer-free individuals. Genome-wide DNA methylation profiling showed a global decrease in methylation in IM compared with normal tissues (median methylation = 0.64 and 0.70 for gastric cardiac IM and normal tissues, respectively). Differential methylation analysis between gastric cardiac IM and normal tissues identified 38,237 differentially methylated probes (DMPs) with a majority of sites showing hypermethylation in IM compared with normal tissues (56.3% vs. 43.7%). Subsequent analysis revealed a significant enrichment of hypermethylated DMPs in promoter and CpG islands (p < 0.001 for both, Pearson χ2 test). For DMPs located in promoter CpG islands showing extreme hypermethylation, the candidate gene with the largest number of DMPs (n = 7) was mapped to HOXA5. Accordingly, mRNA expression of HOXA5 was significantly reduced in IM compared to normal tissue. Our results suggest the implication of alterations in DNA methylation in gastric cardiac IM and highlight that HOXA5 hypermethylation may be a promising epigenetic biomarker, emphasizing the role of aberrant HOXA5 expression in the pathogenesis of gastric cardiac IM.


Health-related quality of life in patients with recurrent pericarditis: results from a phase 2 study of rilonacept.
Impact of recurrent pericarditis (RP) on patient health-related quality of life (HRQoL) was evaluated through qualitative patient interviews and as an exploratory endpoint in a Phase 2 trial evaluating the efficacy and safety of rilonacept (IL-1α/IL-1β cytokine trap) to treat RP. Qualitative interviews were conducted with ten adults with RP to understand symptoms and HRQoL impacts, and the 10-item Patient-Reported Outcomes Measurement Information System Global Health (PROMIS GH) v1.2 was evaluated to determine questionnaire coverage of patient experience. The Phase 2 trial enrolled participants with active symptomatic RP (A-RP, n = 16) and corticosteroid-dependent participants with no active recurrence at baseline (CSD-RP, n = 9). All participants received rilonacept weekly during a 6-week base treatment period (TP) plus an optional 18-week extension period (EP). Tapering of concomitant medications, including corticosteroids (CS), was permitted during EP. HRQoL was assessed using the PROMIS GH, and patient-reported pain and blood levels of c-reactive protein (CRP) were collected at Baseline and follow-up periods. A secondary, descriptive analysis of the Phase 2 trial efficacy results was completed using HRQoL measures to characterize both the impact of RP and the treatment effect of rilonacept. Information from qualitative interviews demonstrated that PROMIS GH concepts are relevant to adults with RP. From the Phase 2 trial, both participant groups showed impacted HRQoL at Baseline (mean PROMIS Global Physical Health [GPH] and Global Mental Health [GMH], were lower than population norm average). In A-RP, GPH/MPH improved by end of base TP and were sustained through EP (similar trends were observed for pain and CRP). Similarly, in CSD-RP, GPH/MPH improved by end of TP and further improved during EP, during CS tapering or discontinuation, without disease recurrence (low pain scores and CRP levels continued during the TP and EP). This is the first study demonstrating impaired HRQoL in RP. Rilonacept treatment was associated with HRQoL improvements using PROMIS GH scores. Maintained/improved HRQoL during tapering/withdrawal of CS without recurrence suggests that rilonacept may provide an alternative to CS. ClinicalTrials.Gov; NCT03980522; 5 June 2019, retrospectively registered; https://clinicaltrials.gov/ct2/show/NCT03980522 .


Informed consent in the intensive care unit: the experiences and expectations of patients and their families.
To describe the awareness of intensive care unit patients and their next of kin (NoK) about invasive procedures and their expectations of informed consent for procedures in the ICU. A written survey of patients and their NoK in a tertiary, university-affiliated ICU, using multiplechoice questions, Likert scales and comments to generate semiquantitative and qualitative data. Fifty-one ICU patients and 69 NoK completed the survey. Inclusion criteria were unplanned ICU admission, ICU length-of-stay > 24 hours, English speaking and competent to consent to participate. Proportion of procedures respondents were aware had occurred during ICU admission; satisfaction with information received; preferred method of receiving information and giving consent; and expectations of when procedural consent is required. Patients and NoK were unaware of many procedures performed during their admission. Respondents correctly identified 49% (95% CI, 45%-53%) of procedures performed during the patient's ICU admission. Despite this, most patients (80%; 95% CI, 69%-91%) and NoK (94%; 95% CI, 89%-100%) were satisfied with information provided about procedures. Over half of respondents (55%; 95% CI, 46%-64%) only expected consent for procedures that were "risky or not routine". About one-quarter (27%; 95% CI, 23%-31%) expected to give consent before every procedure and 15% (95% CI, 11%-18%) expected no procedural consent process. Patients and NoK strongly preferred a verbal rather than written consent process. Our results suggest there is a limited degree of support for routine procedural consent from ICU patients and their NoK.


Kinetic studies on muscle glycogen synthase.
Using the I form of rabbit muscle glycogen synthase essentially free of glycogen, the kinetics and mechanism of action was investigated. No evidence for an exchange between [14C]UDP and UDP-glucose was found. The bisubstrate kinetics of the enzyme for UDP-glucose and glycogen, as well as for UDP-glucose and maltose, was determined. An intersecting pattern in the double reciprocal plot (velocity versus substrate concentration) suggestive of a sequential mechanism (ordered or random) was found in all cases. The K-m for UDP-glucose (45 to 48 mM) was the same with either maltose or glycogen as acceptor. The K-m for maltose (230 mM) and for glycogen (1.5 mug/ml) differed.


Animal model of depression: pharmacological sensitivity of a hedonic deficit.
A reduction in sucrose and saccharine consumption following chronic stress is reported for the rat. This deficit may be related to consummatory deficits seen in endogenous depression. To further examine this state pharmacologically, stressed rats were treated with the antidepressant imipramine. Despite a general absence of appetitive effects (or in some cases mild anorexia) imipramine significantly restored saccharine consumption in a variety of tests. The pharmacological similarity of the deficit to the changes accompanying affective disorders further supports the potential applicability of the chronic stress model.


Amino-acid sequences of the active-site sulfhydryl peptide and other thiol peptides from the cysteine proteinase alpha-clostripain.
One free -SH group in the heavy chain of alpha-clostripain reacts rapidly with N-tosyllysine chloromethyl ketone which inactivates the enzyme. Iodoacetic acid also reacts with the thiol group required for enzyme activity but more slowly. A tryptic peptide containing the reactive sulfhydryl group labelled with iodo[1-14C]acetic acid was isolated and determined to be Gln-Ser-Val-Asp-Leu-Leu-Ala-Phe-Asp-Ala-Cys-Met. All other cysteine peptides were isolated from the trypsin hydrolysate of the [14C]carboxymethylated enzyme. Moreover N-terminal and C-terminal sequences of both chains of alpha-clostripain were determined. The sequences representing 20% of the primary structure of alpha-clostripain are not homologous with either other cysteine proteinases or with any other protein structure known to date.


Evaluation of changes in endolymphatic volume after middle ear pressure therapy for intractable meniere's disease using 3D magnetic resonance imaging.
Nowadays, the endolymphatic space size can be evaluated by 3D-analysis of 3 T-MRI after intravenous injection of gadolinium enhancement. In the present study, to elucidate the relationships between vertigo and endolymphatic hydrops (EH) volume after middle ear pressure therapy (MEPT), we investigated changes in EH volume after MEPT for intractable Meniere's disease (MD) by means of the inner ear MRI (ieMRI) in relation to clinical results. We enrolled 202 successive definite MD cases with intractable vertiginous symptoms from 2015 to 2020, assigning Group-I of MEPT, and Group-II of endolymphatic sac drainage (ELSD). Ninety patients completed the planned 2-year-follow-up, which included assessment of vertigo frequency and changes in EH volume using ieMRI (Group-I/MEPT: n = 40; Group-II/ELSD: n = 50). Two years after surgery, vertigo was completely controlled in 77.5% of patients in Group-I and 90.0% in Group-II. Hearing improved by >10 dB in 7.5% of patients in Group-I and 24.0% in Group-II. ELS ratios were significantly reduced after treatments of Group-I and Group-II only in the vestibule. The obtained results indicate that MEPT as well as ELSD could be a good treatment option for patients with intractable MD.


Human milk as a vector and an indicator of exposure to PCBs and PBDEs: temporal trend of samples collected in Rome.
Thirty-seven polychlorobiphenyl (PCB) congeners and seven polybromodiphenylether (PBDE) congeners were measured in human milk samples collected in Rome between 2005 and 2007. The comparison of results with two previous studies performed in Rome in 1984 and in 2000-2001 indicates a 64% decrease of PCB levels, still in progress; profile differences with time were also evident as lighter congeners are less relevant now; data are in good agreement with recent European studies. PBDE contamination profiles were different in individual samples and a similar variability was observed in data from different countries, suggesting different exposure pathways and profiles.


Compliance with statewide regulations for communication of patients' multidrug-resistant organism and Clostridium difficile status during transitions of care.
In 2014, Oregon implemented an interfacility transfer communication law requiring notification of multidrug-resistant organism status on patient transfer. Based on 2015 and 2016 statewide facility surveys, compliance was 77% and 87% for hospitals, and 67% and 68% for skilled nursing facilities. Methods for complying with the rule were heterogeneous, and fewer than half of all facilities surveyed reported use of a standardized interfacility transfer communication form to assess a patient's multidrug-resistant organism status on transfer.


Delayed Effect of Dendritic Cells Vaccination on Survival in Glioblastoma: A Systematic Review and Meta-Analysis.
Dendritic cell vaccination (DCV) strategies, thanks to a complex immune response, may flare tumor regression and improve patients' long-term survival. This meta-analysis aims to assess the efficacy of DCV for newly diagnosed glioblastoma patients in clinical trials. The study databases, including PubMed, Web of Knowledge, Google Scholar, Scopus, and Cochrane, were searched by two blinded investigators considering eligible studies based on the following keywords: "glioblastoma multiforme", "dendritic cell", "vaccination", "immunotherapy", "immune system", "immune response", "chemotherapy", "recurrence", and "temozolomide". Among the 157 screened, only 15 articles were eligible for the final analysis. Regimens including DCV showed no effect on 6-month progression-free survival (PFS, HR = 1.385, 95% CI: 0.822-2.335, p = 0.673) or on 6-month overall survival (OS, HR = 1.408, 95% CI: 0.882-2.248, p = 0.754). In contrast, DCV led to significantly longer 1-year OS (HR = 1.936, 95% CI: 1.396-2.85, p = 0.001) and longer 2-year OS (HR = 3.670, 95% CI: 2.291-5.879, p = 0.001) versus control groups. Hence, introducing DCV could lead to increased 1 and 2-year survival of patients by 1.9 and 3.6 times, respectively. Antitumor regimens including DCV can effectively improve mid-term survival in patients suffering glioblastoma multiforme (GBM), but its impact emerges only after one year from vaccination. These data indicate the need for more time to achieve an anti-GBM immune response and suggest additional therapeutics, such as checkpoint inhibitors, to empower an earlier DCV action in patients affected by a very poor prognosis.


Genome-wide transcript profiling indicates induction of energy-generating pathways and an adaptive immune response in the liver of sows during lactation.
The present study aimed to explore the lactation-induced changes in hepatic gene expression in sows (Sus scrofa) during lactation. Using a porcine whole-genome microarray a total of 632 differentially expressed genes in the liver of lactating compared to non-lactating sows could be identified. Enrichment analysis revealed that the differentially expressed genes were mainly involved in fatty acid metabolism, pyruvate metabolism, glutathione metabolism, glycine, serine and threonine metabolism, citrate cycle, glycerophospholipid metabolism, PPAR signaling, and focal adhesion. The most striking observation with respect to intermediary metabolism was that genes involved in fatty acid catabolism, the catabolism of gluconeogenic amino acids, the citrate cycle and the respiratory chain were up-regulated in the liver of sows during lactation. With respect to immune response, it could be demonstrated that genes encoding acute phase proteins and genes involved in tissue repair were up-regulated and genes encoding adhesion molecules were down-regulated in the liver of sows during lactation. The results indicate that energy-generating pathways and pathways involved in the delivery of gluconeogenic substrates are induced in sow liver during lactation. The alterations of expression of genes encoding proteins involved in immune response suggest that lactation in sows may cause an adaptive immune response that possibly counteracts hepatic inflammation.


Oviduct development inDrosophila : I. Effects of ovariectomy and genital disc transplantations.
The lateral oviducts ofDrosophila virilis were investigated in normal unoperated adult females, after unilateral and bilateral ovariectomy, and following the transplantation of genital discs. Subsequent to unilateral ovariectomy at larval stages of development, mature adult females exhibited reproductive systems with a free lateral oviduct which appeared somewhat shorter and less distended than a lateral oviduct normally attached to an ovary. Histological examination revealed that such free lateral oviducts have undergone considerable growth and differentiation in the absence of direct ovarian attachment, but exhibited a smaller lumen and more highly folded epithelium. They may be distinguished from attached lateral oviducts by conformational differences and by a possibly lesser size of the epithelial cells. Free lateral oviducts observed among bilaterally ovariectomized and sham-ovariectomized specimens were indistinguishable from those obtained after unilateral ovariectomy. The results are at variance with the previously accepted conclusion that oviduct growth inDrosophila is dependent upon inductive influences emanating from the ovary and directing the outgrowth of the oviducts. Differences in the developmental performance of the oviducts as a function of age at the time of ovariectomy were not evident in the study which included larvae ranging from the second instar to late in the third instar. Transplants of larval female genital discs to other larvae revealed a lesser development of the lateral oviducts than that exhibited by a genital disc developingin situ. A range of oviducal growth which lacked any relation to ovarian influences or to other internal organs of the hosts was obtained. In general, decreased amounts of oviducal muscle were found associated with the transplants.


Thymic neuroendocrine carcinoma (carcinoid): a clinicopathologic study of fourteen cases.
The medical records and histologic documents of 14 patients treated at our institution for a thymic carcinoid tumor were reviewed. There were 3 women and 11 men with an age range from 35 to 71 years. One patient had a multiple endocrine neoplasia syndrome; another had a neurofibromatosis. Twelve tumors were revealed by local symptoms and two were asymptomatic. One patient had Cushing's syndrome that appeared secondarily and was related to metastases. Tumors ranged from 6 to 20 cm and had the characteristic histologic appearance of atypical carcinoid tumor. Immunohistochemical evaluations were done. Tumors were positive for cytokeratin (92%), neuroendocrine markers (100%), and p53 oncoprotein (29%). S-100 protein antibody revealed numerous sustentacular cells in one case. Overall survival was 46% and 31% at 3 and 5 years, respectively. However, all patients died of the disease within 109 months as a result of local progression (n = 5), local relapse (n = 3), distant metastases (n = 8), or a combination of these reasons. Median survival was 71, 30, and 5 months for patients who had total resection (n = 4), partial resection (n = 5), or simple biopsy (n = 4), respectively (p = 0.023). In conclusion, thymic carcinoid tumors can be considered thymic neuroendocrine carcinomas because of their malignant behavior and histologic appearance of atypical carcinoid tumors. Complete surgical resection offers the best hope for long-term survival.


Beginning to Read in Vietnamese: Kindergarten Precursors to First Grade Fluency and Reading Comprehension.
Measures of decoding and oral language have been shown to predict early reading comprehension across a wide variety of languages, though the timeframe and strength of the predictions vary by orthographic depth. This study is the first to examine predictors of early reading in Vietnamese, a transparent orthography of Romanized letters and diacritics. Eighty-two children in Hanoi, Vietnam, completed measures of decoding and oral language in kindergarten (phonological awareness, PA; rapid automatized naming, RAN; receptive and expressive vocabulary) and measures of decoding and reading comprehension in first grade. Average performance at the end of first grade, after just one year of formal instruction, was near ceiling on word reading but more variable on nonword and text reading. Kindergarten PA and RAN (but not vocabulary) predicted first-grade decoding after controlling for maternal education and kindergarten decoding, and PA was a stronger predictor than RAN (10% vs 4% of unique variance). The best predictors of first grade reading comprehension were first-grade decoding (47% of unique variance) and kindergarten expressive vocabulary (4% of unique variance) after controlling for maternal education. Overall, Vietnamese children became accurate and efficient decoders after one year of instruction. Findings from RAN and PA suggest their utility in guiding differentiated instruction on decoding. Kindergarten vocabulary, which differed as a function of maternal education, significantly predicted first-grade comprehension.


Impact of the Paraglottic Space on Voice Production in an MRI-Based Vocal Fold Model.
While the vocal fold is in direct contact anteriorly with the thyroid cartilage, posteriorly the vocal fold connects to the thyroid cartilage through a soft tissue layer in the paraglottic space. Currently the paraglottic space is often neglected in computational models of phonation, in which a fixed boundary condition is often imposed on the lateral surface of the vocal fold. The goal of this study was to investigate the effect of the paraglottic space on voice production in an MRI-based vocal fold model, and how this effect may be counteracted by vocal fold stiffening due to laryngeal muscle activation. Parametric simulation study using an MRI-based computational vocal fold model. The results showed that the presence of the paraglottic space increased the mean and amplitude of the glottal area waveform, decreased the phonation frequency and closed quotient. For the particular vocal fold geometry used in this study, the presence of the paraglottic space also reduced the occurrence of irregular vocal fold vibration. These effects of the paraglottic space became smaller with increasing paraglottic space stiffness and to a lesser degree with vocal fold stiffening. The results suggest that the paraglottic space may be neglected in qualitative evaluations of normal phonation, but needs to be included in simulations of pathological phonation or vocal fold posturing.


Whole mount in situ hybridization and immunohistochemistry for studying retinoic acid signaling in developing amphioxus.
Retinoic acid (RA) is a vitamin A-derived signaling molecule acting during development and in the adult. This chapter provides protocols to characterize the role of RA signaling during development of the invertebrate chordate amphioxus. As sister group to all other chordates and characterized by the most vertebrate-like RA signaling system of all invertebrates, amphioxus is an important model for studying the evolution of RA signaling. Focusing on the development of GABAergic neurons in the amphioxus central nervous system, we provide detailed protocols for maintaining and breeding adult animals, for performing pharmacological treatments of embryos and for analyzing the effects of these treatments by whole mount in situ hybridization and immunohistochemistry coupled to confocal microscopy.


The use of epidemiology, scientific data, and regulatory authority to determine risk factors in cancers of some organs of the digestive system. 6. Pancreatic cancer.
The etiology of pancreatic cancer remains elusive. It predominates in males and in certain ethnic populations (i.e., Polynesians and blacks). Primarily a disease of aging, it is rare in individuals under 40. Genetics is believed to play a very small part if any. While diabetics have higher than expected incidence and mortality rates, several recent studies have indicated that in most cases the diabetes is an early sign of pancreatic cancer rather than a predisposing condition. Other conditions have been infrequently reported in association with pancreatic cancer. None of them have shown a definitive associative pattern and are believed to represent coincidental occurrences. Migrant studies of Japanese immigrants implicate some type of environmental etiology, since pancreatic cancer rates increase dramatically within one to two generations. Studies on atom bomb survivors have demonstrated no link between a single intense dose of radiation and subsequent development of this disease. Workers exposed to low-level radiation have shown an increase in pancreatic cancer, but this result may reflect methodologic problems. Many chemicals have been shown to cause pancreatic cancer in animals, and chemists exhibit higher pancreatic cancer rates than expected. However, the human cases have not been traced to any specific chemical. Many other occupations besides chemistry have shown increased pancreatic cancer rates, but the common factor in these occupations is not obvious. Studies analyzing the relationship of alcohol consumption to pancreatic cancer yield conflicting results. This may be explained if the observed effects are due to a confounder, such as cigarette smoking; to an ingredient other than alcohol contained in alcoholic beverages, such as nitrosamines; or to the immunosuppressive effects of chronic excessive alcohol consumption. Smoking is the risk factor showing the most definitive and consistent results. There is little doubt that it plays an etiologic role, and it probably accounts for the higher incidence of the disease in males. Most other research does not support MacMahon's reported association between coffee drinking and pancreatic cancer. Recent studies have indicated a generally increased fluid intake in pancreatic cancer patients due to a disease-induced disturbance in glucose tolerance function. Such increased fluid intake would tend to manifest itself in the most popular beverage of the country (i.e., coffee in the United States and tea in the United Kingdom). Animal studies have indicated a link between pancreatic cancer and high fat and/or high protein diets as well as raw soybean consumption.(ABSTRACT TRUNCATED AT 400 WORDS)


Randomized Clinical Trials and Real World Prospective Observational Studies on Bevacizumab, PARP Inhibitors, and Immune Checkpoint Inhibitors in the First-Line Treatment of Advanced Ovarian Carcinoma: A Critical Review.
Platinum/paclitaxel-based chemotherapy is able to obtain a clinical response in up to 80% of patients with advanced ovarian carcinoma, but most of them will subsequently develop recurrent disease. Several therapeutic approaches, including prolonged administration of the first-line regimen and the concomitant or sequential addition of a third cytotoxic agent to standard chemotherapy, failed to improve the clinical outcome of patients. In the last years, the implementation of the biological knowledge on ovarian carcinoma and the introduction of bevacizumab (BEV) and poly(ADP-ribose) polymerase inhibitors (PARPi) in first-line treatment have improved patient prognosis. In this review, we have analyzed the randomized clinical trials and real world observational studies on these issues, with the aim to suggest an algorithm for a rational use of BEV and PARPi in patients with newly diagnosed advanced ovarian carcinoma.


Transcriptional characterization of the Notch signaling pathway in rodent multipotent adult progenitor cells.
The Notch signaling pathway is a multifunctional, evolutionarily conserved pathway, which plays an important role in development as well as stem cell biology. Multipotent adult progenitor cells (MAPCs) represent a unique stem cell population, which is capable of differentiating into cell types of the ectodermal, mesodermal and endodermal lineages in vitro, and contribute to most somatic cell types in vivo. Our aim was to characterize the gene expression of Notch signaling elements in rodent MAPCs. We show that transcripts for Notch-receptors, ligands, regulatory molecules of the pathway and the Hairy/Enhancer of Split-1 (HES-1) target gene are present in mouse and rat low-Oct4 MAPCs. We found that mouse Notch3 and rat Notch1 transcripts increased when cells were cultured at high density for 48 to 96 hours. HES-1 and HES-related transcription factor-1 (HERP-1), transcriptional targets of Notch-signaling, were both elicited by immobilized Delta1 ligand. In addition, mRNA for Notch1 and Notch3 was also induced by Notch-signaling, suggesting the presence of regulatory feedback loops. Slight differences between mouse and rat derived MAPCs suggest that the exact function, transcriptional regulation and the fine-tuning of the signal may be species specific. Taken together, we characterized the gene expression profile of the Notch pathway in rodent low-Oct4-MAPCs, and showed that the pathway is functional and can be modulated. Our results provide an additional tool and a further basis for a better understanding of stem cell biology.


[The significance of the preoperative CEA value for the prognosis of rectal cancer].
254 patients were operated for rectal carcinoma in the period from 1980 to 1986. The number of curative procedures, recurrence rate and 5-year-survival rate were dependent on the preoperative CEA levels, which correlated with the tumour stage. Even within tumour stages with enough cases for statistical evaluation the CEA could be established as an independent prognostical variable. Within stage pT2N0M0 recurrence rate for patients with preoperative CEA level below 5 ng/ml was 29% compared to 64% in patients with higher serum levels. Median survival time for the two groups were 65 and 35 months, respectively. In stage of lymph node involvement median survival times of 46 and 25 months were observed for the groups with CEA levels above and below 5 ng/ml. The pattern of recurrence within the two stages seemed to be unaffected by the preoperative CEA level. Patients with preoperative elevated CEA represent a risk group with need of consistent postoperative follow-up and CEA controls.


Serotonin-like immunoreactivity in the epidermal club cells of teleost fishes.
The present immunocytochemical study concerns the distribution of serotonin in the epidermis of three species of teleost fish. Serotonin-like immunoreactivity was found in the club cells of Heteropneustes fossilis and Carapus acus but not in those from the sea eel Conger conger. This study is the first immunocytochemical identification of serotonin in the club cells of teleost epidermis. By comparing data from the literature (Zaccone et al. 1986, 1987, 1988) regarding the occurrence of serotonin and GRP/bombesin in the exocrine sacciform gland cells of piscine skin, it is worthy mentioning here that the serotonin contained in the club cells of the species studied may have the ability to affect the pheromonal or other possible functions of these cells. The presence of serotonin in these systems has been correlated with the capacity of the exocrine glands of fish skin to secrete, ectopically, amine messengers in contrast to those produced eutopically i.e. in the neuron-paraneuron system in some vertebrates (Fujita et al. 1988).


A novel mutation in TNNT3 associated with Sheldon-Hall syndrome in a Chinese family with vertical talus.
Distal arthrogryposis (DA) is a group of rare, clinically and genetically heterogeneous disorders primarily characterized by congenital contractures of the limb joints. Recently, mutations in genes encoding the fast-twitch skeletal muscle contractile myofibers complex, including troponin I2 (TNNI2), troponin T3 (TNNT3), tropomyosine 2 (TPM2), and embryonic myosin heavy chain 3 (MYH3), and the slow-twitch skeletal muscle myosin binding protein C1 (MYBPC1) were confirmed to cause DA1, DA2A, and DA2B. DA2B, or Sheldon-Hall syndrome (SHS; MIM 601680), is intermediate to DA1 and DA2A, or Freeman-Sheldon syndrome (FSS; MIM193700), and shows prominent facial traits. This report describes a Chinese family with SHS over three generations in which all affected individuals showed vertical talus and one demonstrated preauricular tags on the face. Linkage analysis and PCR sequencing revealed a novel substitute mutation at a hot-spot site in TNNT3 (c.187C > T; p.R63C). This mutation was confirmed to cosegregate with the DA phenotype in affected individuals. SIFT and PolyPhen analyses suggest that the mutation is pathogenic. We report this mutation in TNNT3 and speculate that bilateral vertical talus, or severe clubfoot, might be a special characteristic for cases with the TNNT3 R63C mutation.


Conflict resolution: unlocking the key to success.
Delivering the best care possible starts with healthcare providers. If we don't get along,what happens to our patients?


Is laparoscopic liver resection safe for intrahepatic cholangiocarcinoma? A meta-analysis.
The use of laparoscopic liver resection for curative surgery of intrahepatic cholangiocarcinoma (ICC) is not well established. Herein, we perform a meta-analysis to compare the differences between laparoscopic liver resection (LLR) and open liver resection (OLR) for ICC. Multiple electronic databases were searched and 8 relevant studies containing 552 patients treated by LLR and 2320 treated by OLR were identified. The fixed effects and a random-effects model were used to perform a meta-analysis. Compared with OLR, LLR for ICC was associated with less blood transfusion (7.14% versus 17.11%; OR: 0.32; 95% CI 0.15 to 0.71; P < 0.05), higher R0 resection (85.63% versus 74.69%; OR: 1.48; 95% CI 1.13 to 1.95; P < 0.05), shorter length of stay (LOS) (SMD: -0.40; 95% CI -0.80 to 0.00; P = 0.05), less overall morbidities (20% versus 32.69%; OR: 0.50; 95% CI 0.33 to 0.78; P < 0.05), and less death due to tumor recurrence (22.39% versus 35.48%; OR: 0.50; 95% CI 0.29 to 0.86; P <0.05); but LLR was associated with smaller ICC, fewer major hepatectomies, less lymph node (LN) dissection rate, and inferior 5-year overall survival (OS) (P < 0.05). Duration of operation, blood loss, average LN retrieved, LN metastasis, major morbidities, mortality, tumor recurrence, 3-year OS and disease free survival (DFS), and 5-year DFS were comparable (P >0.05). LLR for ICC is in the initial phase of exploration. More evidence is necessary to validate LLR for ICC.


Feasibility of Immunoadsorption plasmapheresis in an infant for ABO-incompatible solid organ transplant-A reality.
Immunoadsorption (IA) plasmapheresis is standard modality for pretransplant desensitization in ABO-incompatible solid organ transplants though technically challenging when considered for an infant or a child less than 10 kg due to non-availability of pediatric immunoadsorption (IA) columns. The major challenge is to maintain hemodynamic stability considering the large extracorporeal circuit volume meant for adults. To our best knowledge after extensive search in acclaimed global medical journals, this is the first successful attempt in an underweight (6 kg) infant of less than 1 year of age using adult size IA Column thus making it a reality. We report an 8-month-old male infant (A positive) of 6 kg with decompensated liver disease secondary to extrahepatic biliary atresia requiring urgent live donor liver transplantation with AB positive donor with significantly elevated pretransplant anti-B IgG/ IgM antibody titers >1:1024. Baby underwent multiple sessions of anti-B immunoadsorption plasmapheresis to lower anti-B IgM / IgG titers using available adult anti-B immunoadsorption column. Postprocedure, the antibody titers reduced to 1:8 (anti-IgG) 1:16 (anti-IgM) followed by successful ABO-incompatible live donor liver transplant (LDLT). Anti-B titers remained in normal range in the immediate and post-transplant period with satisfactory liver functions and no rejection. Immunoadsorption plasmapheresis for ABO-incompatible solid organ transplantation in infants gives desirable results and can be offered to small sized infants using currently available adult sized IA columns when conducted with adequate technical expertise.


Off-label application of intravenous immunoglobulin (IVIG) for treatment of Cogan's syndrome during pregnancy.
We report the case of a woman with Cogan's syndrome concomitant with the wish to have children. After three major flares of the disease that led to unilateral deafness, immunosuppressive therapy with prednisolone and azathioprine was started. Because of the severe side effects, an off-label therapy with intravenous immunoglobulin (IVIG) was initiated, under which our patient has since given birth to three healthy children. To our knowledge this is the first report to describe Cogan's syndrome with multiple successful pregnancies under IVIG treatment.


The stability of inhibited/uninhibited temperament from ages 3 to 11 years in Mauritian children.
Stability of inhibited/uninhibited temperament was assessed using 1,795 Mauritian children tested at ages 3, 8, and 11 years. Children were divided into uninhibited, middle, and inhibited groups at each age based on social behavior. Results indicated that, relative to uninhibited children (1) those inhibited at age 3 obtained larger inhibition scores at age 8 (p < .0001), (2) those inhibited at age 8 obtained larger inhibition scores at age 11 (p < .002), and (3) those remaining inhibited from ages 3 to 8 obtained larger inhibition scores at age 11 (p < .002). Relative to children who changed classification from ages 3 to 8, those remaining inhibited obtained larger inhibition scores (p < .05) and those remaining uninhibited obtained smaller inhibition scores (p < .015) at age 11. Inhibition scores tended to be higher in females by age 11. Results remained regardless of ethnicity. The results provide some support that inhibited/uninhibited temperament remains stable from ages 3 to 8 and may continue to age 11. The results suggest cross-cultural generalizability of these findings with implications regarding the development of anxiety disorders in the Mauritian population.


Increased expression of thioredoxin-1, vascular endothelial growth factor, and redox factor-1 is associated with poor prognosis in patients with liver metastasis from colorectal cancer.
We examined whether the expression of thioredoxin-1 (Trx-1) was associated with patient prognosis after liver resection for metastatic colorectal cancer. Eighty-four patients underwent resection of liver metastases from colorectal cancer, leaving no macroscopic evidence of residual tumor. Immunohistochemical study was performed to evaluate the relation among Trx-1, vascular endothelial growth factor (VEGF), and redox factor-1 (Ref-1) expression and the clinicopathologic characteristics and patient survival. Thirty-seven patients (44.0%) with Trx-1-positive metastases had shorter survival after primary liver resection (P = .0003) than the 47 patients (56.0%) with Trx-1-negative metastases. The percentage VEGF-positive and Ref-1-positive metastases was significantly higher in patients with Trx-1 expression (P = .0009 and .0002, respectively). Multivariate analysis revealed that Trx-1 expression was an independent prognostic factor. Expression of VEGF and Ref-1 is associated with Trx-1 overexpression, which is related to a poor prognosis in patients with liver metastases from colorectal cancer.


Activin A Modulates Inflammation in Acute Pancreatitis and Strongly Predicts Severe Disease Independent of Body Mass Index.
Acute pancreatitis (AP) is a healthcare challenge with considerable mortality. Treatment is limited to supportive care, highlighting the need to investigate disease drivers and prognostic markers. Activin A is an established mediator of inflammatory responses, and its serum levels correlate with AP severity. We hypothesized that activin A is independent of body mass index (BMI) and is a targetable promoter of the AP inflammatory response. We assessed whether BMI and serum activin A levels are independent markers to determine disease severity in a cohort of patients with AP. To evaluate activin A inhibition as a therapeutic, we used a cerulein-induced murine model of AP and treated mice with activin A-specific neutralizing antibody or immunoglobulin G control, both before and during the development of AP. We measured the production and release of activin A by pancreas and macrophage cell lines and observed the activation of macrophages after activin A treatment. BMI and activin A independently predicted severe AP in patients. Inhibiting activin A in AP mice reduced disease severity and local immune cell infiltration. Inflammatory stimulation led to activin A production and release by pancreas cells but not by macrophages. Macrophages were activated by activin A, suggesting activin A might promote inflammation in the pancreas in response to injury. Activin A provides a promising therapeutic target to interrupt the cycle of inflammation and tissue damage in AP progression. Moreover, assessing activin A and BMI in patients on hospital admission could provide important predictive measures for screening patients likely to develop severe disease.


TRFMA: a web-based tool for terminal restriction fragment length polymorphism analysis based on molecular weight.
TRFMA provides a Web environment for analyzing T-RFLP results based on molecular weights of the fragments, rather than the numbers of nucleotides, to increase accuracy. The 16S rRNA data are saved as an XML file containing around 650 sequences (light version) and a MySQL database containing around 50 000 sequences (full version), which are connected to Web server via PHP5 and manipulated on an Internet browser. TRFMA is freely available at http://myamagu.dent.kyushu-u.ac.jp/bioinformatics/trfma/index.html and can be downloaded from the same site.


G-protein signaling in iNOS gene expression.
Heterotrimeric G proteins are the molecular switches in the receptor-mediated transmembrane signaling system. Inducible nitric oxide synthase (iNOS) is inducible by a variety of inflammatory stimuli, which leads to vascular hyporeactivity. In this chapter, the system to study the cell signaling pathways downstream of the GPCR coupling to G proteins is described for the study of iNOS gene expression. The cellular signaling pathways by which ligand induces iNOS may serve as the pharmacological targets for preventing or treating vascular hyporeactivity.


Gastrostomy for infants with severe epidermolysis bullosa simplex in neonatal intensive care.
Severe epidermolysis bullosa simplex (EBS sev) is a rare genodermatosis characterized by congenital generalized blistering and mucosal involvement. Increased needs and decreased intake quickly lead to nutritional imbalance. Enteral nutrition support is proposed, but classical nasogastric tubes are not well tolerated in these patients and gastrostomy is preferred. To report the experience with EBS sev in neonatal units of French reference centers for gastrostomy. In this retrospective multicentric study, we included all patients with EBS sev who had gastrostomy placement before age 9 months during neonatal care hospitalization. Nine infants (5 males/4 females) with severe skin and mucosal involvement were included. A gastrostomy was decided, at an early age (mean 3.7 months, range 1.4 to 8 months) in infants with mean weight 4426 g (range 3500 to 6000 g). Techniques used were endoscopy with the pull technique for 5 infants and surgery under general anesthesia for 4. Main complications were local but resolved after treatment. All infants gained weight after gastrostomy. The mean withdrawal time (n = 7) for the gastrostomy was 35.8 months (range 10.5 months to 6.5 years). Seven children had persistent oral disorders. Gastrostomy in infants with EBS sev can be necessary in neonatal intensive care units. Both surgical and endoscopic pull techniques seem efficient, with good tolerance.


The impact of an intervention to improve intrapartum maternal vital sign monitoring and reduce alarm fatigue.
The infrastructure of many labor and delivery units in the United States may dispose clinicians to overuse continuous and automated maternal physiological monitors. Overmonitoring low-risk patients can negatively affect patient care, primarily through generating alarm fatigue. Given the national attention to reducing alarm fatigue across healthcare settings and the concern for vital sign monitoring overuse on our labor and delivery unit, this quality improvement study aimed to evaluate vital sign monitoring patterns and alarm rates, and nursing experiences of alarm fatigue, before and after implementing a vital sign monitoring guideline for low-risk obstetrical patients. This was a quality improvement study conducted on the labor and delivery unit of an urban, academic, tertiary hospital. The lack of guidance for maternal vital sign assessment in low-risk patients was identified as a potential safety challenge. A vital sign guideline was developed with multidisciplinary input, followed by a pre-post-implementation study evaluating vital sign volume and alarm rates. Total vital signs and alarm rates for all patients delivered during designated calendar days were assessed as a rate of vital signs per patient and compared across baseline, peri-intervention, and follow-up periods. Data were examined in p-type statistical process control charts and with time-series analysis. Patient characteristics and severe maternal morbidity, as a balancing metric, were compared across periods. Nursing perceptions of vital sign monitoring and experience of alarm fatigue were assessed via survey before and after implementation of the guideline. A total of 35 individual 24-hour periods were evaluated with regard to vital sign and alarm volume. There was a decrease in vital signs per patient from a mean of 208.34 to 135.46 (incidence rate ratio, 0.65) and in alarms per patient from a mean of 14.31 to 10.51 (incidence rate ratio, 0.73) after implementation, with no difference in severe maternal morbidity. There were 85 total respondents to the nursing surveys, and comparison of modified task-load index scores before and after implementation demonstrated overall lower scores in the postperiod, although these were not statistically significant. Introducing a maternal vital sign guideline for low-risk patients on the labor and delivery unit decreased vital signs measured as well as alarms, which may ultimately reduce alarm fatigue. This strategy should be considered on labor and delivery units widely to improve patient safety and optimize outcomes.


C-H Bond Activation by Iridium(III) and Iridium(IV) Oxo Complexes.
Oxidation of an iridium(III) oxo precursor enabled the structural, spectroscopic, and quantum-chemical characterization of the first well-defined iridium(IV) oxo complex. Side-by-side examination of the proton-coupled electron transfer thermochemistry revealed similar driving forces for the isostructural oxo complexes in two redox states due to compensating contributions from H+ and e- transfer. However, C-H activation of dihydroanthracene revealed significant hydrogen tunnelling for the distinctly more basic iridium(III) oxo complex. Our findings complement the growing body of data that relate tunnelling to ground state properties as predictors for the selectivity of C-H bond activation.


Systematic approach to electrode insertion in the ossified cochlea.
Ossification of the fluid spaces of the cochlea occurs often in candidates for cochlear implantation, especially children. When noted before surgery on computerized tomography, ossification previously was thought to contraindicate cochlear implantation because of possible mechanical obstruction and uncertainty about the level of function that could be achieved by stimulating an ossified cochlea. However, during the preceding 6 years, techniques have been developed that permit implantation in ossified cochleas. We present a systematic approach that has been developed to treat the three clinically important categories of cochlear ossification: round window niche obliteration, inferior segment obstruction, and upper segment obstruction. Case reports are presented for each of these three drill-out procedures, demonstrating results often similar to those expected for implantation of the nonossified cochlea.


A smooth start to a new age. Helping women cope with the menopause.
The menopause is a natural event in women's lives, but its associations with loss of fertility and its side-effects often lead to it being seen in a negative light. Nurses can help women control the side-effects so they may learn to see it as a positive change.


Conformational analysis of the anomeric forms of sophorose, laminarabiose, and cellobiose using MM3.
Relaxed-residue energy maps based on the MM3 force-field were computed for relative orientations of the pyranosyl rings of sophorose, laminarabiose, and cellobiose, respectively the (1----2)-beta-; (1----3)-beta-; and (1----4)-beta-linked D-glucosyl disaccharides. Sixteen starting conformations of the rotatable exocyclic side-groups were considered for each molecule. All of the energy surfaces have two intersecting low-energy troughs and illustrate the importance of exo-anomeric effects in determining disaccharide conformation. Local minima were found by relaxed minimization without restriction. The energy surfaces of these disaccharides are very similar to the energy surfaces of their corresponding 6-methyltetrahydropyran analogues. There is good agreement between disaccharide structures having minimal MM3 energy and those found by crystallography.


SVIP regulates Z variant alpha-1 antitrypsin retro-translocation by inhibiting ubiquitin ligase gp78.
Alpha-1 antitrypsin deficiency (AATD) is an inherited disorder characterized by early-onset emphysema and liver disease. The most common disease-causing mutation is a single amino acid substitution (Glu/Lys) at amino acid 342 of the mature protein, resulting in disruption of the 290-342 salt bridge (an electrophoretic abnormality defining the mutation [Z allele, or ZAAT]), protein misfolding, polymerization, and accumulation in the endoplasmic reticulum of hepatocytes and monocytes. The Z allele causes a toxic gain of function, and the E3 ubiquitin ligase gp78 promotes degradation and increased solubility of endogenous ZAAT. We hypothesized that the accumulation of ZAAT is influenced by modulation of gp78 E3 ligase and SVIP (small VCP-interacting protein) interaction with p97/VCP in ZAAT-expressing hepatocytes. We showed that the SVIP inhibitory effect on ERAD due to overexpression causes the accumulation of ZAAT in a human Z hepatocyte-like cell line (AT01). Overexpression of gp78, as well as SVIP suppression, induces gp78-VCP/p97 interaction in AT01 cells. This interaction leads to retro-translocation of ZAAT and reduction of the SVIP inhibitory role in ERAD. In this context, overexpression of gp78 or SVIP suppression may eliminate the toxic gain of function associated with polymerization of ZAAT, thus providing a potential new therapeutic approach to the treatment of AATD.


Gait disorders.
Gait disorders are frequently accompanied by loss of balance and falls, and are a common cause of disability, particularly among the elderly. In many cases the cause is multifactorial, involving both neurologic and nonneurologic systems. Physical therapy and training, coupled with pharmacologic and surgical therapy, can usually provide some improvement in ambulation, which translates into better quality of life. More research is needed on the mechanisms of gait and its disorders as well as on symptomatic therapies. Better understanding of the pathophysiology of gait disorders should lead to more specific, pathogenesis-targeted therapies.


Metabolic effects of carbon monoxide in relation to atherogenesis.
The acute and chronic effects of carbon monoxide (CO) are reviewed. Direct effects of CO-induced hypoxia on arterial metabolism may facilitate deposition of cholesterol in the intimal surface and exacerbate existing atherosclerosis. Other studies indicate that CO may alter hepatic metabolism so as to inhibit both the secretion of very low density lipoproteins and the clearance of chylomicron remnants. The former change would diminish and the latter enhance atherogenesis. These findings are considered and avenues of future investigation suggested.


The effect of short-term corticosteroid incubation on the alloactivating and antigen-presenting capacity of human epidermal Langerhans cells.
The effects of short-term incubation with hydrocortisone and methylprednisolone on the alloactivating capacity and the ability of human epidermal Langerhans cells and peripheral blood monocytes to induce antigen (PPD)-specific T-lymphocyte responses were investigated. The Langerhans cells and the monocytes were treated with different concentrations of the steroids before being added to allogeneic T lymphocytes, and before, after or simultaneously with, pulsing with PPD. Neither steroid inhibited the alloactivating capacity of the Langerhans cells, but a dose-dependent reduction of the stimulatory capacity of antigen-pulsed Langerhans cells and monocytes was obtained with methylprednisolone. Hydrocortisone had inhibitory effects only at high concentration. The inhibition appeared to be due to suppression of the presentation of PPD in immunogeneic form to sensitized T cells.


Building an efficient factory: where is pre-rRNA synthesized in the nucleolus?
The subnucleolar structure that is involved in rDNA transcription has been controversial. A report by Koberna et al. (2002)(this issue, page 743) adds significant weight toward the idea that dense fibrillar components (DFCs)**Abbreviations used in this paper: DFC, dense fibrillar component; FC, fibrillar center; GC, granular component; Pol I, polymerase I. and fibrillar center (FC)/DFC borders are the sites of pre-rRNA synthesis.


First report of Hemicriconemoides litchi associated with Piper sarmentosum and revision of the genus Hemicriconemoides in Vietnam.
The taxonomic status of Hemicriconemoides litchi, H. mangiferae, and H. strictathecatus has been in debates for many years. In this study, a population of H. litchi collected from the rhizosphere of Piper sarmentosum is characterized using a combination of morphology and molecular data. Our results are in agreement with other authors to maintain the validity of H. litchi and provide sequences of H. litchi with correct names on GenBank. A revision on the genus Hemicriconemoides in Vietnam is also discussed. The taxonomic status of Hemicriconemoides litchi, H. mangiferae, and H. strictathecatus has been in debates for many years. In this study, a population of H. litchi collected from the rhizosphere of Piper sarmentosum is characterized using a combination of morphology and molecular data. Our results are in agreement with other authors to maintain the validity of H. litchi and provide sequences of H. litchi with correct names on GenBank. A revision on the genus Hemicriconemoides in Vietnam is also discussed.


Factors associated with relative muscle strength in patients with type 2 diabetes mellitus.
Some patients with type 2 diabetes mellitus (T2DM) experience decreased mobility associated with lower relative muscle strength (normalized with muscle mass). This study aimed to identify factors predicting relative muscle strength of patients with T2DM assessed at regular clinical visits. A total of 144 T2DM patients underwent fasting blood drawing (determining white blood cell count [WBC], diabetic kidney disease [DKD], and glycated hemoglobin [HbA1c]) and the assessment of body composition, diabetic peripheral neuropathy (DPN), activity level, and muscle strength (grip, knee extensor, and ankle plantar flexor strength). One-way ANOVA and multiple regression models were used to identify factors associated with the relative muscle strength. Our data showed that age, diabetes duration, fat percentage, WBC, DPN, and DKD were negatively associated with the relative muscle strength. Specifically, a greater WBC was associated with lower relative muscle strength of both distal and proximal muscle groups of extremities after the adjustment of other associated factors. DPN was associated with lower relative strength of the distal muscle groups of extremities. WBC may be used as a marker of inflammation, and greater count, even within the normal range, is negatively associated with the relative muscle strength in patients with T2DM.


Treatment of transient bone marrow oedema of the hip--a comparative study.
Between 1990 and 2000, we treated 43 patients with transient bone marrow oedema of the hip. Five were treated with nonsteroidal antiinflammatory drugs (NSAIDs) and limited weight bearing, and 38 by core decompression followed by limited weight bearing. At follow-up 2-10 years later, all patients were assessed by a structured interview as well as the Harris hip score (HHS) and the Western Ontario and MacMaster Universities Osteoarthritis Index (WOMAC). Both groups reached the same clinical outcome (HHS and WOMAC). Core decompression enabled a significantly faster recovery. There were no complications, but progression to avascular necrosis was seen in both groups. Core decompression induced fast pain relief, making it the preferable treatment.


Activation of spinal macrophage-inducible C-type lectin induces mechanical allodynia and microglial activation in rats.
Macrophage-inducible C-type lectin (Mincle), a pattern recognition receptor, is a critical component of the innate immune system that is involved in the pathogenesis of chronic pain. Previous studies have reported the expression of Mincle in neuronal and glial cells of the brain, but its expression and role in pain processing at the spinal level remain to be determined. The current study was performed to identify Mincle in the spinal cord and to investigate the effect of Mincle activation on spinal sensitization. Most Mincle immunoreactivity was localized within the grey matter and the dorsal and ventral horns of the lumbar spinal cord in naïve rats. A single intrathecal (i.t.) injection of trehalose-6,6-dibehenate (TDB), a Mincle ligand, induced mechanical allodynia. Immunoreactivity to Mincle and Iba-1 in the spinal cord significantly increased after i.t. injection of TDB. Mechanical allodynia was attenuated by daily i.t. injection of minocycline. However, double immunofluorescence revealed that Mincle co-localizes with NeuN (neurons), but not with Iba-1 (microglia) or GFAP (astrocytes). In conclusion, we found that Mincle was present in spinal cord neurons, but not microglia or astrocytes, and may play a role in microglia-induced spinal sensitization.


DW14006 as a direct AMPKα1 activator improves pathology of AD model mice by regulating microglial phagocytosis and neuroinflammation.
Alzheimer's disease (AD) is a progressively neurodegenerative disease with typical hallmarks of amyloid β (Aβ) plaque accumulation, neurofibrillary tangle (NFT) formation and neuronal death extension. In AD brain, activated microglia phagocytose Aβ and neuronal debris, but also aggravate inflammation stress by releasing inflammatory factors and cytotoxins. Improving microglia on Aβ catabolism and neuroinflammatory intervention is thus believed to be a promising therapeutic strategy for AD. AMP-activated protein kinase (AMPK) is highly expressed in microglia with AMPKα1 being tightly implicated in neuroinflammatory events. Since indirect AMPKα1 activators may cause side effects with undesired intracellular AMP/ATP ratio, we focused on direct AMPKα1 activator study by exploring its potential function in ameliorating AD-like pathology of AD model mice. Here, we reported that direct AMPKα1 activator DW14006 (2-(3-(7-chloro-6-(2'-hydroxy-[1,1'-biphenyl]-4-yl)-2-oxo-1,2-dihydroquinolin-3-yl)phenyl)acetic acid) effectively improved learning and memory impairments of APP/PS1 mice, and the underlying mechanisms have been intensively investigated. DW14006 reduced amyloid plaque deposition by promoting microglial o-Aβ42 phagocytosis and ameliorated innate immune response by polarizing microglia to an anti-inflammatory phenotype. It selectively enhanced microglial phagocytosis of o-Aβ42 by upgrading scavenger receptor CD36 through AMPKα1/PPARγ/CD36 signaling and suppressed inflammation by AMPKα1/IκB/NFκB signaling. Together, our work has detailed the crosstalk between AMPKα1 and microglia in AD model mice, and highlighted the potential of DW14006 in the treatment of AD.


Ten-year follow-up of a randomized trial of pravastatin in heart transplant patients.
Outcomes from this trial's first year data demonstrated significant benefit in heart transplant patients treated with pravastatin in cholesterol levels, survival, rejection with hemodynamic compromise, the development of cardiac allograft vasculopathy, and decreased natural killer cell cytotoxicity. Other heart transplant studies have shown similar benefit. We now report the 10-year follow-up of this study. Ninety-seven heart transplant recipients were randomized to pravastatin (n = 47) or no pravastatin (n = 50) within 2 weeks after surgery both in combination with cyclosporine and corticosteroids. Ten-year outcomes include survival, cholesterol levels, and development of cardiac allograft vasculopathy documented by coronary angiography. Forty-two percent of the control patients crossed over to pravastatin treatment during the second year of the study, and 81% of the control patients were eventually placed on statin therapy by the 10-year follow-up. The control group had subsequent low and comparable cholesterol levels in Years 2 to 10 of the study compared with the patients originally randomized to pravastatin. Intent-to-treat analysis demonstrated that the pravastatin group compared with control had increased 10-year survival (68% vs 48%, p = 0.026). The 10-year freedom from angiographic cardiac allograft vasculopathy and/or death in the pravastatin group was significantly greater compared with the control group (43% vs 20%, p = 0.009). The 10-year follow-up of this study suggests that the use of pravastatin in heart transplant patients maintains survival benefit and appears to reduce the development of cardiac allograft vasculopathy.


Cross-linking mass spectrometry discovers, evaluates, and corroborates structures and protein-protein interactions in the human cell.
Significant recent advances in structural biology, particularly in the field of cryoelectron microscopy, have dramatically expanded our ability to create structural models of proteins and protein complexes. However, many proteins remain refractory to these approaches because of their low abundance, low stability, or-in the case of complexes-simply not having yet been analyzed. Here, we demonstrate the power of using cross-linking mass spectrometry (XL-MS) for the high-throughput experimental assessment of the structures of proteins and protein complexes. This included those produced by high-resolution but in vitro experimental data, as well as in silico predictions based on amino acid sequence alone. We present the largest XL-MS dataset to date, describing 28,910 unique residue pairs captured across 4,084 unique human proteins and 2,110 unique protein-protein interactions. We show that models of proteins and their complexes predicted by AlphaFold2, and inspired and corroborated by the XL-MS data, offer opportunities to deeply mine the structural proteome and interactome and reveal mechanisms underlying protein structure and function.


Correspondence regarding Schwend and Gustafsson, "False positives in MALDI-TOF detection of ERbeta in mitochondria".
Recently, Schwend and Gustafsson tried to use the MALDI-TOF methods to confirm one of the results reported by Yang et al., which provided definitive evidences to demonstrate the localization of estrogen receptor beta (ERbeta) in the mitochondria of multiple cell types, using immunocytochemistry, immunoblot, and proteomic approaches. Analysis of the data with the MASCOT database algorithm provided no evidence for the presence of ERbeta in the mouse live mitochondria, in which very low ERbeta expression has been detected in their own report. On the other hand, our MALDI-TOF analysis using human heart mitochondrial protein has identified 7 and 8 sequences that could be potentially from ERbeta and ERbeta3, respectively, but not from ATP synthases. Further, none of the sequences identified by us as those of ERbeta and ERbeta3 shares m/z targeted by Schwend and Gustafsson in their measurements. Therefore, the claim by Gustafsson's laboratory about false positives in MALDI-TOF detection of ERbeta in mitochondria has no relevance to our report.


[Impact of antithrombotic treatment and geriatric syndromes in octogenarians with atrial fibrillation and ischaemic heart disease. Atrial Fibrillation and Ischemic Heart Disease in the Elderly].
The prevalence of atrial fibrillation (AF) and ischaemic heart disease (IHC) increases with age. They coexist in up to 20% of octogenarian patients, a situation that poses a therapeutic challenge. Trials that have addressed this scenario, which included a low percentage of octogenarians, showed that double therapy (single antiplatelet + anticoagulation) compared to triple therapy (double antiplatelet + anticoagulation) was associated with less bleeding events, especially with direct oral anticoagulants. These studies did not have sufficient power to detect differences in ischaemic events. On the other hand, prevalent characteristics in the elderly, such as geriatric syndromes, were not assessed in these studies, and are not usually evaluated in clinical practice. Accordingly, their prognostic impact remains unknown in this clinical context. Observational, prospective, and multicentre study that will include patients ≥ 80 years with AF and IHC in Spain. Baseline characteristics and geriatric syndromes will be assessed, as well as the choice of antithrombotic treatment. The primary endpoint is cardiovascular and overall mortality at one and three years follow-up. This study will assess both characteristics and prognosis of octogenarian patients with AF and IHC in Spain, the factors involved in the choice of antithrombotic treatment, and the incidence of ischaemic and haemorrhagic events during the short- and long-term follow-up. This study will contribute to improve the knowledge in terms of safety and efficacy of the different therapeutic options in older patients with AF and IHC, as well as their prognostic impact.


Correlation of enhancement degree on contrast-enhanced ultrasound with histopathology of carotid plaques and serum high sensitive C-reactive protein levels in patients undergoing carotid endarterectomy.
This study was undertaken to investigate the correlation of the enhancement degree on contrast-enhanced ultrasound (CEUS) with the histopathology of carotid plaques and the serum high sensitive C-reactive protein (hs-CRP) levels in patients undergoing carotid endarterectomy (CEA). Carotid CEUS was performed preoperatively in 115 patients who would undergo CEA, and the enhancement degree of the carotid plaques was evaluated by both the visual semiquantitative analysis and the quantitative time-intensity curve analysis. Serum hs-CRP levels were detected using the particle-enhanced immunoturbidimetric assay also before the operation. Additionally, the carotid plaque samples were subjected to histopathological examination postoperatively. The density of neovessels and the number of macrophages in the plaques were assessed by immunohistochemistry. The results showed that among the 115 patients, grade 0 plaque contrast enhancement was noted in 35 patients, grade 1 in 48 patients and grade 2 in 32 patients. The degree of plaque enhancement, the density of neovessels, the number of macrophages, and the hs-CRP levels were highest in the grade 2 patients. Correlation analysis showed that the enhancement degree of the carotid plaques was closely related to the immunohistochemical parameters of the plaques and the serum hs-CRP levels. It was suggested that the carotid plaque enhancement on CEUS can be used to evaluate the vulnerability of carotid plaques.


Plant biomes demonstrate that landscape resilience today is the lowest it has been since end-Pleistocene megafaunal extinctions.
Resilient landscapes have helped maintain terrestrial biodiversity during periods of climatic and environmental change. Identifying the tempo and mode of landscape transitions and the drivers of landscape resilience is critical to maintaining natural systems and preserving biodiversity given today's rapid climate and land use changes. However, resilient landscapes are difficult to recognize on short time scales, as perturbations are challenging to quantify and ecosystem transitions are rare. Here we analyze two components of North American landscape resilience over 20,000 years: residence time and recovery time. To evaluate landscape dynamics, we use plant biomes, preserved in the fossil pollen record, to examine how long a biome type persists at a given site (residence time) and how long it takes for the biome at that site to reestablish following a transition (recovery time). Biomes have a median residence time of only 230-460 years. Only 64% of biomes recover their original biome type, but recovery time is 140-290 years. Temperatures changing faster than 0.5°C per 500 years result in much reduced residence times. Following a transition, biodiverse biomes reestablish more quickly. Landscape resilience varies through time. Notably, short residence times and long recovery times directly preceded the end-Pleistocene megafauna extinction, resulting in regional destabilization, and combining with more proximal human impacts to deliver a one-two punch to megafauna species. Our work indicates that landscapes today are once again exhibiting low resilience, foreboding potential extinctions to come. Conservation strategies focused on improving both landscape and ecosystem resilience by increasing local connectivity and targeting regions with high richness and diverse landforms can mitigate these extinction risks.


HDAC7 promotes the oncogenicity of nasopharyngeal carcinoma cells by miR-4465-EphA2 signaling axis.
HDAC7 plays a crucial role in cancers, and is the main drug target of several HDAC inhibitors. However, the role and mechanism of HDAC7 in nasopharyngeal carcinoma (NPC) are still unclear. In this study, we observed that HDAC7 was significantly upregulated in the NPC tissues relative to normal nasopharyngeal mucosa (NNM) tissues, HDAC7 expression levels were positively correlated with NPC progression and negatively correlated with patient prognosis, and HDAC7 knockdown dramatically inhibited the in vitro proliferation, migration, and invasion of NPC cells, and the growth of NPC xenografts in mice, indicating the HDAC7 promotes the oncogenicity of NPC. Mechanistically, HDAC7 promoted the in vitro proliferation, migration, and invasion of NPC cells by upregulating EphA2, in which miR-4465 mediated HDAC7-regulating EphA2, a direct target gene of miR-4465. We further showed that miR-4465 was significantly downregulated in the NPC tissues relative to NNM tissues, and inhibited the in vitro proliferation, migration, and invasion of NPC cells by targeting EphA2 expression. Moreover, we observed that the expressions of HDAC7, miR-4465, and EphA2 in NPC tissues were correlated. The results suggest that HDAC7 promotes the oncogenicity of NPC by downregulating miR-4465 and subsequently upregulating EphA2, highlighting HDAC7 as a potential therapeutic target for NPC.


Chemo-physical properties of asbestos bodies in human lung tissues studied at the nano-scale by non-invasive, label free x-ray imaging and spectroscopic techniques.
In the lungs, asbestos develops an Fe-rich coating (Asbestos Body, AB) that becomes the actual interface between the foreign fibers and the host organism. Conventional approaches to study ABs require an invasive sample preparation that can alter them. In this work, a novel combination of x-ray tomography and spectroscopy allowed studying unaltered lung tissue samples with chrysotile and crocidolite asbestos. The thickness and mass density maps of the ABs obtained by x-ray tomography were used to derive a truly quantitative elemental analysis from scanning x-ray fluorescence spectroscopy data. The average mass density of the ABs is compatible with that of highly loaded ferritin, or hemosiderin. The composition of all ABs analyzed was similar, with only minor differences in the relative elemental fractions. Silicon concentration decreased in the core-to-rim direction, indicating a possible partial dissolution of the inner fiber. The Fe content in the ABs was higher than that possibly contained in chrysotile and crocidolite. This finding opens two opposite scenarios, the first with Fe coming from the fiber bulk and concentrating on the surface as long as the fiber dissolves, the second where the Fe that takes part to the formation of the AB originates from the host organism Fe-pool.


Occurrence of bla genes encoding carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter baumannii from Intensive Care Unit in a tertiary care hospital.
ICU shows increasing incidence of infection associated with the use of invasive procedures for the diagnostic purpose as well as the indiscriminate use of antibiotics. Pseudomonas aeruginosa and Acinetobacter species are "very successful" pathogen and the emergence of the Metallo-β-Lactamases (MBL) is becoming a therapeutic challenge. To isolate the Nonfermenting Gram negative bacilli from the ICU samples. To identify the metallo betalactamase producers and to detect the bla gene presence among the Pseudomonas aeruginosa and Acinetobacter baumannii. The Nonfermenting Gram negative bacilli isolates from the ICU samples were taken over for 5 years (2009-2014) in a tertiary care hospital. The isolates of Pseudomonas species and Acinetobacter species were confirmed by API analyser and processed according to standard procedures. Detection of the MBL producers were done by E strip method and subjected for bla gene detection by PCR method. In our study a total of 195 isolates of NFGNB were obtained from various ICU. Of these MBL producers, 26 % were Pseudomonas aeruginosa and 25 % were Acinetobacter baumannii. The subtypes of blaVIM MBL producing P.aeruginosa were 26%. The predominant gene coding for MBL activity in A.baumannii were found to be blaOXA gene 11.9%. The gene accession numbers were KF975367, KF975372. We have to control the development and dissemination of these superbugs among the ICU's.


Development of a ginkgo biloba fingerprint chromatogram with UV and evaporative light scattering detection and optimization of the evaporative light scattering detector operating conditions.
A fingerprint chromatogram of a standardized Ginkgo biloba extract is developed on a monolithic silica column using a ternary gradient containing water, iso-propanol and tetrahydrofuran. For the detection, UV and evaporative light scattering (ELS) detectors are used, the latter allowing detection of the poor UV absorbing compounds as ginkgolides (A-C and J) and bilobalide in the extract. The complementary information between the UV and ELS fingerprint is evaluated. The ELS detector used in this study can operate in an impactor 'on' or 'off' mode. For each mode, the operating conditions such as the nebulizing gas flow rate, the drift tube temperature and the gain are optimized by use of three-level screening designs to obtain the best signal-to-noise (S/N) ratio in the final ELS fingerprint chromatogram. In both impactor modes, very similar S/N ratios are obtained for the nominal levels of the design. However, optimization of the operating conditions resulted, for both impactor modes, in a significant increase in S/N ratios compared to the initial evaluated conditions, obtained from the detector software.


Inhibin concentration in the culture media of human oocyte-cumulus-corona cell complexes is not related to subsequent embryo cleavage.
We have demonstrated an in vitro secretion of inhibin by human OCCC, not related to subsequent egg cleavage. A putative signal, released by OCCC, that could modulate inhibin secretion by granulosa cells has not been discovered. Inhibin secretion by OCCC has no predictive value concerning the further fertilization and cleavage of the oocytes.


Ruania alkalisoli sp. nov., Isolated from Saline-Alkaline Soil.
A Gram-positive, strictly aerobic, ivory-colored, rod-shaped bacterial strain, designated RN3S43T, was isolated from saline-alkaline soil, in Tumd Right Banner, Inner Mongolia, China. Strain RN3S43T grew at 10-40 °C (optimum 30 °C), pH 6.0-10.0 (optimum pH 9.0), and 0-12.5% NaCl (optimum 2-4%). It was positive to oxidase, catalase, urease, and nitrate reductase. The methyl red and Voges-Proskauer tests were negative. The phylogenetic trees based on the 16S rRNA gene sequences and genome both showed that strain RN3S43T clustered with Ruania alba YIM 93306 T and shared 95.5% and < 95.0% of 16S rRNA gene similarities with R. alba YIM 93306 T and all the other type strains. MK-8 was the major respiratory quinone. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, and an unidentified lipid were the major polar lipids. The major fatty acids were anteiso-C15:0 and iso-C15:0. The genome consisted of a 4,448,708-bp circular chromosome, with a G+C content of 68.2%, predicting 3,911 coding sequence genes, 44 tRNA genes and two rRNA operons. The average nucleotide identity (ANI), amino acid identity (AAI), and digital DNA-DNA hybridization (dDDH) values of strain RN3S43T to R. alba YIM 93306T were 79.0%, 79.2%, and 23.4%, respectively. The results of phylogenetic, physiological, and phenotypic tests allowed the discrimination of strain RN3S43T from its phylogenetic relatives. Ruania alkalisoli sp. nov. is therefore proposed, and the type strain is RN3S43T (=CGMCC 1.18652T = KCTC 49471T).


Quantitative expansion of structural genomic alterations in the spectrum of neuroendocrine lung carcinomas.
The pathogenesis and interrelationships of neuroendocrine lung carcinomas are not well understood. Tissue macro-arrays prepared from surgical resection specimens from 35 patients with typical carcinoid (TC), six with atypical carcinoid (AC), 13 with large cell neuroendocrine carcinoma (LCNEC), and 15 with small cell lung carcinoma (SCLC) were investigated by fluorescence in situ hybridization (FISH) and immunohistochemistry. Hybridizations with locus-specific DNA probes demonstrated a high incidence of deletion for the tumour suppressor genes p53 and retinoblastoma (Rb), and for the oncogene cyclin D1, comparable in all carcinoma types. Similarly, an increase of DNA copy number for the Her-2/neu and c-myc oncogenes was noted in all neoplasms. A more detailed quantitative analysis of the results, however, demonstrated increasing numbers of cells harbouring these genomic alterations, from low-grade TC to highly malignant SCLC, with the exception of cyclin D1 deletion. Mutations of the p53 and Rb genes, as assayed by immunohistochemical studies, were observed at high incidence in high-grade carcinomas, compared with a low incidence in the low-grade carcinomas. Conversely, in all carcinoma types, neither membrane-bound Her-2/neu nor nuclear cyclin D1 was detected. It is concluded that structural genomic alterations are frequent in neuroendocrine lung carcinomas and that their occurrence may be underestimated by immunohistochemical studies alone. The quantitative expansion of the Rb, p53, c-myc, and Her-2/neu alterations towards high-grade carcinomas suggests common pathogenetic mechanisms in the spectrum of these neoplasms.


Image Classification of Amazon Parrots by Deep Learning: A Potentially Useful Tool for Wildlife Conservation.
Parrots play a crucial role in the ecosystem by performing various roles, such as consuming the reproductive structures of plants and dispersing plant seeds. However, most are threatened because of habitat loss and commercial trade. Amazon parrots are one of the most traded and illegally traded parrots. Therefore, monitoring their wild populations and global trade is crucial for their conservation. However, monitoring wild populations is becoming more challenging because the manual analysis of large-scale datasets of images obtained from camera trap methods is labor-intensive and time consuming. Monitoring the wildlife trade is difficult because of the large quantities of wildlife trade. Amazon parrots can be difficult to identify because of their morphological similarity. Object detection models have been widely used for automatic and accurate species classification. In this study, to classify 26 Amazon parrot species, 8 Single Shot MultiBox Detector models were assessed. Among the eight models, the DenseNet121 model showed the highest mean average precision at 88.9%. This model classified the 26 Amazon parrot species at 90.7% on average. Continuous improvement of deep learning models classifying Amazon parrots may support monitoring wild populations and the global trade of these species.


Cineangiography in the diagnosis of aortic dissection.
To assess the ability of biplane cineangiography in the diagnostic evaluation of acute dissection of the aorta, 20 patients with acute dissection were studied within 24 hours of surgery or autopsy, or both. Biplane large film aortic angiography was performed in 11 patients (Group I) and biplane aortic cineangiography in 9 (Group II). The morphology of the aortic valve was defined precisely in 5 (50 percent) of 10 patients in Group I and in all 9 patients (100 percent) in Group II (p less than 0.02). Aortic regurgitation weas diagnosed in all patients in both groups in whom it was present (p = not significant). Intimal tears were localized in 5 (50 percent) of 10 patients in Group I and in 8 (89 percent) of 9 in Group II (p less than 0.07). Intimal flaps were not identified angiographically in three patients in Group I and were identified in four patients (100 percent) in Group II (p less than 0.01). The presence of retrograde dissection was established in three (38 percent) of eight patients in Group I and in four (100 percent) of four patients in Group II (p less than 0.05). There was no difference in the ability to identify a nonclotted false lumen between cine and large film angiography. It is concluded that in addition to improved diagnostic capabilities, technical advantages make cineangiography a good alternative to large film angiography in the diagnostic evaluation of patients with acute dissection of the aorta.


Enhanced profiling and quantification of ginsenosides from mountain-cultivated ginseng and comparison with garden-cultivated ginseng.
Panax ginseng can be generally divided into mountain-cultivated ginseng (MCG) and garden-cultivated ginseng (GCG). The market price of MCG is significantly higher than that of GCG. However, the chemical compositions of MCG and the differences from GCG remained unclear. In this study, an integrated strategy combing an offline two-dimensional liquid chromatography separation, LTQ-orbitrap dual mode acquisition, and Q-trap full quantification/quasi-quantification was proposed to explore and compare the chemical compositions of MCG. Consequently, 559 ginsenosides were characterized, among which 437 ginsenosides were in-depth characterized with α-chain and β-chain annotated. Subsequently, enhanced quantification of 213 ginsenosides was conducted in 57 batches of MCG and GCG. Ginsenosides were found more abundant in MCG than GCG. In addition, 25-year-old MCG could be distinctly differentiated from 15/20-year-old MCG. This strategy facilitated the enhanced profiling and comparison of ginsenosides, improved the quality control tactics of MCG and provided a reference approach for other ginseng related products.


The burden of loneliness: Implications of the social determinants of health during COVID-19.
This study sought to examine if mental health issues, namely depression and anxiety symptoms, and loneliness were experienced differently according to various demographic groups during the COVID-19 pandemic (i.e., a societal stressor). An online survey, comprising demographic questions and questionnaires on depression, anxiety and loneliness symptoms, was distributed in Canada during the height of social distancing restrictions during the COVID-19 pandemic. Respondents (N=661) from lower income households experienced greater anxiety, depression and loneliness. Specifically, loneliness was greater in those with an annual income <$50,000/yr versus higher income brackets. Younger females (18-29yr) displayed greater anxiety, depressive symptoms and loneliness than their male counterparts; this difference did not exist among the other age groups (30-64yr, >65yr). Moreover, loneliness scores increased with increasing depression and anxiety symptom severity category. The relationship between loneliness and depression symptoms was moderated by gender, such that females experienced higher depressive symptoms when encountering greater loneliness. These data identify younger females, individuals with lower income, and those living alone as experiencing greater loneliness and mental health challenges during the height of the pandemic in Canada. We highlight the strong relationship between loneliness, depression and anxiety, and emphasize increased vulnerability among certain cohorts.


Constraint on electromagnetic acceleration of highest energy cosmic rays.
The energetics of electromagnetic acceleration of ultrahigh-energy cosmic rays (UHECRs) is constrained both by confinement of a particle within an acceleration site and by radiative energy losses of the particle in the confining magnetic fields. We demonstrate that the detection of approximately 3 x 10(20) eV events is inconsistent with the hypothesis that compact cosmic accelerators with high magnetic fields can be the sources of UHECRs. This rules out the most popular candidates, namely spinning neutron stars, active galactic nuclei (AGNs). Galaxy clusters and, perhaps, AGN radio lobes and gamma-ray burst blast waves remain the only possible (although not very strong) candidates for UHECR acceleration sites. Our analysis places no limit on linear accelerators. With the data from the future Auger experiment one should be able to answer whether a conventional theory works or some new physics is required to explain the origin of UHECRs.


VSIG4 ameliorates intestinal inflammation through inhibiting macrophages NLRP3 inflammasome and pyroptosis.
Inflammatory bowel disease (IBD) is one of the most common diseases in the digestive system related to aberrant inflammation. V-set and immunoglobulin domain-containing 4 (VSIG4), a type I transmembrane receptor exclusively expressed in a subset of tissue-resident macrophages, has been reported to exert anti-inflammatory activity in immune-related diseases, which has been not explored in IBD yet. This study aims to explore the role and the potential mechanism of VSIG4 in IBD. Clinical samples were obtained from IBD patients and were examined by immunohistochemical staining. THP-1 cells were differentiated into macrophages, and then stimulated with IL-4 plus IL-13 or LPS to induce pro-inflammatory (M1) or anti-inflammatory (M2) phenotype. Cell transfection was conducted to overexpress VSIG4. Western blot and immunofluorescence assays were performed to assess NLRP3 inflammasome- and pyroptosis-related proteins. Cytokines were measured using ELISA. A cell co-culture model of Caco-2 cells and VSIG4-mediated macrophages were established. Cell viability and apoptosis was examined by CCK-8 and flow cytometry assays, respectively. VSIG4 was downregulated in IBD and was negatively correlated with NLRP3 inflammasome. M1 macrophages exhibited higher levels of NLRP3 inflammasome, pyroptosis and inflammatory response than M2 macrophages, while VSIG4 overexpression efficiently reversed these changes in M1 macrophages. In addition, VSIG4 overexpression partly abolished M1 macrophages-induced cell viability loss, inflammatory response, apoptosis and pyroptosis in Caco-2 cells. Collectively, VSIG4 might alleviate intestinal inflammation through regulating M1/M2 macrophages, providing novel insights for the treatment of human IBD.


An alternatingly amphiphilic, resistance-resistant antimicrobial oligoguanidine with dual mechanisms of action.
The increasing number of infections caused by multi-drug resistance (MDR) bacteria is an omen of a new global challenge. As one of the countermeasures under development, antimicrobial peptides (AMPs) and AMP mimics have emerged as a new family of antimicrobial agents with high potential, due to their low resistance generation rate and effectiveness against MDR bacterial strains resulted from their membrane-disrupting mechanism of action. However, most reported AMPs and AMP mimics have facially amphiphilic structures, which may lead to undesired self-aggregation and non-specific binding, as well as increased cytotoxicity toward mammalian cells, all of which put significant limits on their applications. Here, we report an oligomer with the size of short AMPs, with both hydrophobic carbon chain and cationic groups placed on its backbone, giving an alternatingly amphiphilic structure that brings better selectivity between mammalian and bacterial cell membranes. In addition, the oligomer shows affinity toward DNA, thus it can utilize bacterial DNA located in the vulnerable nucleoid as the second drug target. Benefiting from these designs, the oligomer shows higher therapeutic index and synergistic effect with other antibiotics, while its low resistance generation rate and effectiveness on multi-drug resistant bacterial strains can be maintained. We demonstrate that this alternatingly amphiphilic, DNA-binding oligomer is not only resistance-resistant, but is also able to selectively eliminate bacteria at the presence of mammalian cells. Importantly, the oligomer exhibits good in vivo activity: it cleans all bacteria on Caenorhabditis elegans without causing apparent toxicity, and significantly improves the survival rate of mice with severely infected wounds in a mice excision wound model study.


The assay of the activity of protein kinase C with the synthetic oligopeptide substrate designed for histone kinase II.
The activity of histone kinase II was determined on the basis of its ability to phosphorylate the nonapeptide Ala-Ala-Ala-Ser-Phe-Lys-Ala-Lys-Lys-amide designed previously as a specific substrate for this enzyme. Histone kinase II was purified from calf thymus extract by DEAE-cellulose chromatography followed by hydroxylapatite chromatography and high-performance liquid chromatography on a Protein Analysis column (I-125). The Mr value of histone kinase II estimated by the latter method was 50,000-55,000, but several observations indicated that histone kinase II was a product of a proteolytic process. Since the substrate specificity determinants for histone kinase II known from our previous investigations are very similar to those for protein kinase C, it was presumable that histone kinase II was the proteolytic fragment of protein kinase C. Therefore, the nonapeptide was tested as a substrate for protein kinase C prepared from rabbit brain extract by DEAE-cellulose chromatography. The activity of histone kinase II was also detected in brain extract. Histone kinase II was eluted from the DEAE-cellulose in the known position of the proteolytic fragment of protein kinase C. The nonapeptide Ala-Ala-Ala-Ser-Phe-Lys-Ala-Lys-Lys-amide proved to be a better substrate than H1 histone for the detection of the activity of protein kinase C because it was not phosphorylated by the cAMP-dependent protein kinase and the Vmax of protein kinase C was about one order of magnitude higher with the peptide than with H1 histone. The apparent Km of protein kinase C for the peptide was identical with that of histone kinase II (0.2 mM).


Mapping of predictors of the disengagement of the descending inhibitory pain modulation system in fibromyalgia: an exploratory study.
The main symptoms of fibromyalgia comprise diffuse pain, disability, depressive symptoms, catastrophizing, sleep disruption and fatigue, associated with dysfunction of the descending pain-modulating system (DPMS). We aimed to identify patterns of main symptoms of fibromyalgia and neuroplasticity biomarkers (i.e. brain-derived neurotrophic factor (BDNF) and S100B protein) in non-responders to the conditioned pain modulation task (CPM-task) induced by immersion of hand in cold water (0-1°C). Furthermore, we evaluated if these patterns predict responsiveness to CPM-task. This cross-sectional study included 117 women with fibromyalgia ((n = 60) non-responders and (n = 57) responders), with age ranging from 30 to 65 years old. We analysed changes in numerical pain scale (NPS-10) during the CPM-task using a standardized protocol. A hierarchical multivariate logistic regression analysis was used to construct a propensity score-adjusted index to identify non-responders compared to responders to CPM-task. The following variables were retained in the models: analgesic use four or more times per week, heat pain threshold (HPT), poor sleep quality, pain catastrophizing, serum levels of BDNF, number of psychiatric diagnoses and the impact of symptoms of fibromyalgia on quality of life. Receiver operator characteristics (ROC) analysis showed non-responders can be discriminated from responders by a composite index of more frequent symptoms of fibromyalgia and neuroplasticity markers (area under the curve (AUC) = 0.83, sensitivity = 100% and specificity = 98%). Patterns of fibromyalgia symptoms and neuroplasticity markers may be helpful to predict responsiveness to the CPM-task which might help personalize treatment and thereby contribute to the care of patients with fibromyalgia.


A simplified acute kidney injury predictor following transcatheter aortic valve implantation: ACEF score.
Transcatheter aortic valve implantation (TAVI) is an effective, less invasive treatment alternative for symptomatic severe aortic stenosis (AS). Acute kidney injury (AKI) following TAVI is a common complication and is associated with worse outcomes. The age, creatinine, ejection fraction (ACEF) score is a simple scoring method, including only three parameters: age, creatinine, and ejection fraction (EF). The score was well established in predicting AKI after coronary interventions. We aimed to evaluate whether this simple scoring method, ACEF, may predict a development of AKI in patients who underwent TAVI. A total of 173 consecutive patients with symptomatic severe AS who underwent TAVI were included retrospectively. The primary endpoint of the study was the development of AKI. Study population was divided into two groups according to the presence of AKI. The ACEF score was calculated with the formula: age/EF + 1 (if baseline creatinine >2 mg/dl). Twenty-nine patients developed AKI. The median (interquartile range) ACEF score was 1.36 (1.20-1.58). The ACEF score was found to be an independent predictor of AKI (P <0.001). The ACEF score ≥1.36 predicted AKI development with a sensitivity of 96.6% and specificity of 58.8%. Moreover, hypertension, hemoglobin levels, contrast volume, and aortic valve area (AVA) were found to be independent predictors of AKI. Our study revealed that the ACEF score was an independent predictor of AKI. A simple and objective score might be very useful in predicting AKI development in patients undergoing TAVI.


Dummy run and conformity indices in the ongoing EORTC low-grade glioma trial 22033-26033: First evaluation of quality of radiotherapy planning.
Early assessment of radiotherapy (RT) quality in the ongoing EORTC trial comparing primary temozolomide versus RT in low-grade gliomas. RT plans provided for dummy cases were evaluated and compared against expert plans. We analysed: (1) tumour and organs-at-risk delineation, (2) geometric and dosimetric characteristics, (3) planning parameters, compliance with dose prescription and Dmax for OAR (4) indices: RTOG conformity index (CI), coverage factor (CF), tissue protection factor (PF); conformity number (CN = PF x CF); dose homogeneity in PTV (U). Forty-one RT plans were evaluated. Only two (5%) centres were requested to repeat CTV-PTV delineations. Three (7%) plans had a significant under-dosage and dose homogeneity in one deviated > 10%. Dose distribution was good with mean values of 1.5, 1, 0.68, and 0.68 (ideal values = 1) for CI, CF, PF, and CN, respectively. CI and CN strongly correlated with PF and they correlated with PTV. Planning with more beams seems to increase PTV(Dmin), improving CF. U correlated with PTV(Dmax). Preliminary results of the dummy run procedure indicate that most centres conformed to protocol requirements. To quantify plan quality we recommend systematic calculation of U and either CI or CN, both of which measure the amount of irradiated normal brain tissue.


Maternal and neonatal essential fatty acid status in phospholipids: an international comparative study.
To investigate whether the steady decline in the maternal essential fatty acids (EFA) status during pregnancy observed in Dutch pregnant women is a local or general phenomenon. The EFA status was measured during uncomplicated, singleton pregnancy of healthy women from the Netherlands, Hungary, Finland, England and Ecuador. In addition, the EFA status of their neonates were measured at birth. Fatty acid profiles were analyzed in phospholipids isolated from maternal plasma and from umbilical plasma and cord vessel walls. Considerable differences between these centers were observed in the maternal EFA levels and EFA status indexes. However, the change in the absolute as well as relative amounts of the EFAs followed a similar course in the five populations during pregnancy. The neonatal EFA profiles reflected the differences found in maternal plasma during pregnancy and shortly after delivery. Comparable correlations were found, particularly, between the neonatal and the maternal n-3 fatty acids in the participating groups. It seems that the reduction in maternal EFA status during pregnancy is a general phenomenon, and is largely independent of differences in dietary habits and ethnic origin. Since the lowest values for certain maternal EFAs in a given country were significantly higher than the highest value of these EFAs throughout pregnancy in other countries, the functional implications of the pregnancy-associated reduction in the maternal EFA status for the fetal and neonatal development is not obvious and needs to be further elucidated.


Immune Profiling Uncovers Memory T-Cell Responses with a Th17 Signature in Cancer Patients with Previous SARS-CoV-2 Infection Followed by mRNA Vaccination.
It is unclear whether patients with cancer present inherently impaired responses to COVID-19 and vaccination due to their treatments, neoplastic diseases or both. To address this question, immune profiling was performed in three cohorts of healthy donors and oncologic patients: infected with SARS-CoV-2, BNT162b2-vaccinated, and with previous COVID-19 disease and subsequently vaccinated. Cancer patients showed good antibody responses to vaccination, but poor induction of T-cell responses towards the S protein when compared to infection. Following natural infection, the major targets for T-cells were the SARS-CoV-2 structural proteins M and S, but not the N protein. Similar to antibody titers, the T-cell responses quickly decayed after six months post-vaccination. Significant memory T-cell expansion was observed in vaccinated donors only if previously diagnosed with COVID-19 before undergoing vaccination. Oncologic patients with previous COVID-19 followed by vaccination exhibited potent IL-17+ CD4 and CD8 T-cell responses and elevated numbers of circulating neutrophils in peripheral blood.